CN119119321A - Sea cucumber polysaccharide with anti-colitis effect and application thereof - Google Patents
Sea cucumber polysaccharide with anti-colitis effect and application thereof Download PDFInfo
- Publication number
- CN119119321A CN119119321A CN202411612593.7A CN202411612593A CN119119321A CN 119119321 A CN119119321 A CN 119119321A CN 202411612593 A CN202411612593 A CN 202411612593A CN 119119321 A CN119119321 A CN 119119321A
- Authority
- CN
- China
- Prior art keywords
- sea cucumber
- polysaccharide
- cucumber polysaccharide
- intestinal
- formula
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 150000004676 glycans Chemical class 0.000 title claims abstract description 89
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 88
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 88
- 241000251511 Holothuroidea Species 0.000 title claims abstract description 54
- 230000000694 effects Effects 0.000 title abstract description 19
- 206010009887 colitis Diseases 0.000 title abstract description 9
- 150000002772 monosaccharides Chemical class 0.000 claims abstract description 16
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 claims abstract description 12
- PNNNRSAQSRJVSB-SLPGGIOYSA-N Fucose Natural products C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)C=O PNNNRSAQSRJVSB-SLPGGIOYSA-N 0.000 claims abstract description 11
- SHZGCJCMOBCMKK-DHVFOXMCSA-N L-fucopyranose Chemical compound C[C@@H]1OC(O)[C@@H](O)[C@H](O)[C@@H]1O SHZGCJCMOBCMKK-DHVFOXMCSA-N 0.000 claims abstract description 11
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 claims abstract description 10
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 4
- 239000000203 mixture Substances 0.000 claims description 17
- 206010009900 Colitis ulcerative Diseases 0.000 claims description 14
- 201000006704 Ulcerative Colitis Diseases 0.000 claims description 14
- 238000000034 method Methods 0.000 claims description 14
- 239000000843 powder Substances 0.000 claims description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 8
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 claims description 8
- YMKDRGPMQRFJGP-UHFFFAOYSA-M cetylpyridinium chloride Chemical compound [Cl-].CCCCCCCCCCCCCCCC[N+]1=CC=CC=C1 YMKDRGPMQRFJGP-UHFFFAOYSA-M 0.000 claims description 8
- 208000022559 Inflammatory bowel disease Diseases 0.000 claims description 7
- 238000002360 preparation method Methods 0.000 claims description 7
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 6
- 238000011097 chromatography purification Methods 0.000 claims description 6
- 238000001556 precipitation Methods 0.000 claims description 6
- OVRNDRQMDRJTHS-CBQIKETKSA-N N-Acetyl-D-Galactosamine Chemical compound CC(=O)N[C@H]1[C@@H](O)O[C@H](CO)[C@H](O)[C@@H]1O OVRNDRQMDRJTHS-CBQIKETKSA-N 0.000 claims description 5
- MBLBDJOUHNCFQT-UHFFFAOYSA-N N-acetyl-D-galactosamine Natural products CC(=O)NC(C=O)C(O)C(O)C(O)CO MBLBDJOUHNCFQT-UHFFFAOYSA-N 0.000 claims description 5
- AEMOLEFTQBMNLQ-WAXACMCWSA-N alpha-D-glucuronic acid Chemical compound O[C@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-WAXACMCWSA-N 0.000 claims description 5
- 108090000790 Enzymes Proteins 0.000 claims description 4
- 102000004190 Enzymes Human genes 0.000 claims description 4
- 239000003153 chemical reaction reagent Substances 0.000 claims description 4
- 230000003544 deproteinization Effects 0.000 claims description 4
- 230000007071 enzymatic hydrolysis Effects 0.000 claims description 4
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 claims description 4
- 229940088598 enzyme Drugs 0.000 claims description 4
- 239000008187 granular material Substances 0.000 claims description 4
- 238000013268 sustained release Methods 0.000 claims description 4
- 239000012730 sustained-release form Substances 0.000 claims description 4
- 239000003826 tablet Substances 0.000 claims description 4
- 208000011231 Crohn disease Diseases 0.000 claims description 3
- 108090000526 Papain Proteins 0.000 claims description 3
- 239000004365 Protease Substances 0.000 claims description 3
- 229940055729 papain Drugs 0.000 claims description 3
- 235000019834 papain Nutrition 0.000 claims description 3
- 108091005658 Basic proteases Proteins 0.000 claims description 2
- 102000004142 Trypsin Human genes 0.000 claims description 2
- 108090000631 Trypsin Proteins 0.000 claims description 2
- 239000002775 capsule Substances 0.000 claims description 2
- 238000000502 dialysis Methods 0.000 claims description 2
- 239000002552 dosage form Substances 0.000 claims description 2
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 2
- 239000012588 trypsin Substances 0.000 claims description 2
- 238000004042 decolorization Methods 0.000 claims 1
- 238000010612 desalination reaction Methods 0.000 claims 1
- 230000000968 intestinal effect Effects 0.000 abstract description 28
- 241001213462 Holothuria leucospilota Species 0.000 abstract description 16
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 abstract description 9
- 230000037149 energy metabolism Effects 0.000 abstract description 9
- WHBMMWSBFZVSSR-UHFFFAOYSA-N 3-hydroxybutyric acid Chemical compound CC(O)CC(O)=O WHBMMWSBFZVSSR-UHFFFAOYSA-N 0.000 abstract description 8
- MSWZFWKMSRAUBD-UHFFFAOYSA-N beta-D-galactosamine Natural products NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 abstract description 8
- 229940116298 l- malic acid Drugs 0.000 abstract description 8
- UIUJIQZEACWQSV-UHFFFAOYSA-N succinic semialdehyde Chemical compound OC(=O)CCC=O UIUJIQZEACWQSV-UHFFFAOYSA-N 0.000 abstract description 8
- 229930182830 galactose Natural products 0.000 abstract description 5
- 229940097043 glucuronic acid Drugs 0.000 abstract description 5
- MSWZFWKMSRAUBD-IVMDWMLBSA-N 2-amino-2-deoxy-D-glucopyranose Chemical compound N[C@H]1C(O)O[C@H](CO)[C@@H](O)[C@@H]1O MSWZFWKMSRAUBD-IVMDWMLBSA-N 0.000 abstract description 4
- WQZGKKKJIJFFOK-PHYPRBDBSA-N alpha-D-galactose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-PHYPRBDBSA-N 0.000 abstract description 4
- 229960002442 glucosamine Drugs 0.000 abstract description 4
- 208000030159 metabolic disease Diseases 0.000 abstract description 4
- 230000000813 microbial effect Effects 0.000 abstract description 4
- 230000001105 regulatory effect Effects 0.000 abstract description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 3
- 208000035475 disorder Diseases 0.000 abstract description 3
- 238000005516 engineering process Methods 0.000 abstract description 3
- 206010059866 Drug resistance Diseases 0.000 abstract description 2
- 208000030453 Drug-Related Side Effects and Adverse reaction Diseases 0.000 abstract description 2
- 230000007774 longterm Effects 0.000 abstract description 2
- 238000004519 manufacturing process Methods 0.000 abstract description 2
- JCZPMGDSEAFWDY-MGCNEYSASA-N (2r,3s,4s,5r)-2,3,4,5,6-pentahydroxyhexanamide Chemical compound NC(=O)[C@H](O)[C@@H](O)[C@@H](O)[C@H](O)CO JCZPMGDSEAFWDY-MGCNEYSASA-N 0.000 abstract 1
- AEMOLEFTQBMNLQ-AQKNRBDQSA-N D-glucopyranuronic acid Chemical compound OC1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-AQKNRBDQSA-N 0.000 abstract 1
- 241000244126 Holothuria nobilis Species 0.000 description 15
- 241000700159 Rattus Species 0.000 description 13
- 239000002207 metabolite Substances 0.000 description 12
- 239000000047 product Substances 0.000 description 9
- 239000000523 sample Substances 0.000 description 9
- TVZRAEYQIKYCPH-UHFFFAOYSA-N 3-(trimethylsilyl)propane-1-sulfonic acid Chemical compound C[Si](C)(C)CCCS(O)(=O)=O TVZRAEYQIKYCPH-UHFFFAOYSA-N 0.000 description 8
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 8
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 7
- 238000004458 analytical method Methods 0.000 description 7
- 230000035622 drinking Effects 0.000 description 7
- 239000003814 drug Substances 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 7
- 230000002550 fecal effect Effects 0.000 description 7
- 230000001965 increasing effect Effects 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 6
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Chemical compound CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 6
- 102000004889 Interleukin-6 Human genes 0.000 description 6
- 108090001005 Interleukin-6 Proteins 0.000 description 6
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 239000007864 aqueous solution Substances 0.000 description 6
- 229960001927 cetylpyridinium chloride Drugs 0.000 description 6
- 239000003094 microcapsule Substances 0.000 description 6
- 238000000746 purification Methods 0.000 description 6
- 239000012086 standard solution Substances 0.000 description 6
- 238000006467 substitution reaction Methods 0.000 description 6
- MSWZFWKMSRAUBD-GASJEMHNSA-N 2-amino-2-deoxy-D-galactopyranose Chemical compound N[C@H]1C(O)O[C@H](CO)[C@H](O)[C@@H]1O MSWZFWKMSRAUBD-GASJEMHNSA-N 0.000 description 5
- 241001468155 Lactobacillaceae Species 0.000 description 5
- 241001465754 Metazoa Species 0.000 description 5
- 229940079593 drug Drugs 0.000 description 5
- 244000005709 gut microbiome Species 0.000 description 5
- 238000004128 high performance liquid chromatography Methods 0.000 description 5
- 238000000926 separation method Methods 0.000 description 5
- QAOWNCQODCNURD-UHFFFAOYSA-L sulfate group Chemical group S(=O)(=O)([O-])[O-] QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 229920001661 Chitosan Polymers 0.000 description 4
- 206010061218 Inflammation Diseases 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- IAJILQKETJEXLJ-QTBDOELSSA-N aldehydo-D-glucuronic acid Chemical compound O=C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)C(O)=O IAJILQKETJEXLJ-QTBDOELSSA-N 0.000 description 4
- 238000001460 carbon-13 nuclear magnetic resonance spectrum Methods 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 238000001212 derivatisation Methods 0.000 description 4
- 239000002662 enteric coated tablet Substances 0.000 description 4
- 210000003608 fece Anatomy 0.000 description 4
- 239000000499 gel Substances 0.000 description 4
- 230000004054 inflammatory process Effects 0.000 description 4
- 238000004255 ion exchange chromatography Methods 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- KBOPZPXVLCULAV-UHFFFAOYSA-N mesalamine Chemical compound NC1=CC=C(O)C(C(O)=O)=C1 KBOPZPXVLCULAV-UHFFFAOYSA-N 0.000 description 4
- 229960004963 mesalazine Drugs 0.000 description 4
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 4
- 150000003214 pyranose derivatives Chemical class 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 4
- 238000001228 spectrum Methods 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 239000007939 sustained release tablet Substances 0.000 description 4
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 3
- 238000001026 1H--1H correlation spectroscopy Methods 0.000 description 3
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 3
- 108020004414 DNA Proteins 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 3
- 229920005654 Sephadex Polymers 0.000 description 3
- 239000012507 Sephadex™ Substances 0.000 description 3
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 3
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 230000003321 amplification Effects 0.000 description 3
- 239000011230 binding agent Substances 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 229910052799 carbon Inorganic materials 0.000 description 3
- 238000005238 degreasing Methods 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 238000010828 elution Methods 0.000 description 3
- 235000019441 ethanol Nutrition 0.000 description 3
- 239000000945 filler Substances 0.000 description 3
- 239000000796 flavoring agent Substances 0.000 description 3
- 235000013355 food flavoring agent Nutrition 0.000 description 3
- 210000001035 gastrointestinal tract Anatomy 0.000 description 3
- 210000002490 intestinal epithelial cell Anatomy 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000002808 molecular sieve Substances 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- URGAHOPLAPQHLN-UHFFFAOYSA-N sodium aluminosilicate Chemical compound [Na+].[Al+3].[O-][Si]([O-])=O.[O-][Si]([O-])=O URGAHOPLAPQHLN-UHFFFAOYSA-N 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- 239000011550 stock solution Substances 0.000 description 3
- 238000012916 structural analysis Methods 0.000 description 3
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid group Chemical group S(O)(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 3
- 238000001551 total correlation spectroscopy Methods 0.000 description 3
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 2
- GOZMBJCYMQQACI-UHFFFAOYSA-N 6,7-dimethyl-3-[[methyl-[2-[methyl-[[1-[3-(trifluoromethyl)phenyl]indol-3-yl]methyl]amino]ethyl]amino]methyl]chromen-4-one;dihydrochloride Chemical compound Cl.Cl.C=1OC2=CC(C)=C(C)C=C2C(=O)C=1CN(C)CCN(C)CC(C1=CC=CC=C11)=CN1C1=CC=CC(C(F)(F)F)=C1 GOZMBJCYMQQACI-UHFFFAOYSA-N 0.000 description 2
- 241000208140 Acer Species 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- FERIUCNNQQJTOY-UHFFFAOYSA-M Butyrate Chemical compound CCCC([O-])=O FERIUCNNQQJTOY-UHFFFAOYSA-M 0.000 description 2
- 102000004127 Cytokines Human genes 0.000 description 2
- 108090000695 Cytokines Proteins 0.000 description 2
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 2
- 229920002271 DEAE-Sepharose Polymers 0.000 description 2
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- 240000001008 Dimocarpus longan Species 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- 235000000235 Euphoria longan Nutrition 0.000 description 2
- 229920002683 Glycosaminoglycan Polymers 0.000 description 2
- 108010085256 HLP-1 polypeptide Proteins 0.000 description 2
- 238000011993 High Performance Size Exclusion Chromatography Methods 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- 240000007472 Leucaena leucocephala Species 0.000 description 2
- 235000010643 Leucaena leucocephala Nutrition 0.000 description 2
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 2
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 2
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 2
- 239000000853 adhesive Substances 0.000 description 2
- 230000001070 adhesive effect Effects 0.000 description 2
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 2
- 244000052616 bacterial pathogen Species 0.000 description 2
- 238000005452 bending Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 210000001072 colon Anatomy 0.000 description 2
- 230000000875 corresponding effect Effects 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000001962 electrophoresis Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 230000002496 gastric effect Effects 0.000 description 2
- 238000005570 heteronuclear single quantum coherence Methods 0.000 description 2
- 239000000413 hydrolysate Substances 0.000 description 2
- 239000001341 hydroxy propyl starch Substances 0.000 description 2
- 235000013828 hydroxypropyl starch Nutrition 0.000 description 2
- 230000002757 inflammatory effect Effects 0.000 description 2
- 238000002329 infrared spectrum Methods 0.000 description 2
- 230000010354 integration Effects 0.000 description 2
- 238000004811 liquid chromatography Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 2
- 230000002503 metabolic effect Effects 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 235000021096 natural sweeteners Nutrition 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 239000006041 probiotic Substances 0.000 description 2
- 235000018291 probiotics Nutrition 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 238000011002 quantification Methods 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 238000002791 soaking Methods 0.000 description 2
- 239000001632 sodium acetate Substances 0.000 description 2
- 235000017281 sodium acetate Nutrition 0.000 description 2
- 239000007974 sodium acetate buffer Substances 0.000 description 2
- 239000000600 sorbitol Substances 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 230000019635 sulfation Effects 0.000 description 2
- 238000005670 sulfation reaction Methods 0.000 description 2
- 239000006188 syrup Substances 0.000 description 2
- 235000020357 syrup Nutrition 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 239000000811 xylitol Substances 0.000 description 2
- 235000010447 xylitol Nutrition 0.000 description 2
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 2
- 229960002675 xylitol Drugs 0.000 description 2
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- 238000004701 1H-13C HSQC Methods 0.000 description 1
- 238000012584 2D NMR experiment Methods 0.000 description 1
- WUBBRNOQWQTFEX-UHFFFAOYSA-N 4-aminosalicylic acid Chemical compound NC1=CC=C(C(O)=O)C(O)=C1 WUBBRNOQWQTFEX-UHFFFAOYSA-N 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- 101100029848 Arabidopsis thaliana PIP1-2 gene Proteins 0.000 description 1
- 241001678559 COVID-19 virus Species 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- 241000711573 Coronaviridae Species 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 102000053602 DNA Human genes 0.000 description 1
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 1
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 1
- 208000007342 Diabetic Nephropathies Diseases 0.000 description 1
- 206010061818 Disease progression Diseases 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 238000005033 Fourier transform infrared spectroscopy Methods 0.000 description 1
- 238000001157 Fourier transform infrared spectrum Methods 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 208000034826 Genetic Predisposition to Disease Diseases 0.000 description 1
- 241000258195 Holothuria Species 0.000 description 1
- SHZGCJCMOBCMKK-JFNONXLTSA-N L-rhamnopyranose Chemical compound C[C@@H]1OC(O)[C@H](O)[C@H](O)[C@H]1O SHZGCJCMOBCMKK-JFNONXLTSA-N 0.000 description 1
- PNNNRSAQSRJVSB-UHFFFAOYSA-N L-rhamnose Natural products CC(O)C(O)C(O)C(O)C=O PNNNRSAQSRJVSB-UHFFFAOYSA-N 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 241000736262 Microbiota Species 0.000 description 1
- 229920000881 Modified starch Polymers 0.000 description 1
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 229920000331 Polyhydroxybutyrate Polymers 0.000 description 1
- 108020004682 Single-Stranded DNA Proteins 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 229960004909 aminosalicylic acid Drugs 0.000 description 1
- 239000003708 ampul Substances 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 238000005571 anion exchange chromatography Methods 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 238000000149 argon plasma sintering Methods 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000000035 biogenic effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 229960000074 biopharmaceutical Drugs 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical class 0.000 description 1
- 239000000679 carrageenan Substances 0.000 description 1
- 229920001525 carrageenan Polymers 0.000 description 1
- 235000010418 carrageenan Nutrition 0.000 description 1
- 229940113118 carrageenan Drugs 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 125000003636 chemical group Chemical group 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- RNFNDJAIBTYOQL-UHFFFAOYSA-N chloral hydrate Chemical compound OC(O)C(Cl)(Cl)Cl RNFNDJAIBTYOQL-UHFFFAOYSA-N 0.000 description 1
- 229960002327 chloral hydrate Drugs 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000008951 colonic inflammation Effects 0.000 description 1
- 201000002758 colorectal adenoma Diseases 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 238000005100 correlation spectroscopy Methods 0.000 description 1
- 239000003246 corticosteroid Substances 0.000 description 1
- 229960001334 corticosteroids Drugs 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000011033 desalting Methods 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 208000033679 diabetic kidney disease Diseases 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000000378 dietary effect Effects 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 210000002249 digestive system Anatomy 0.000 description 1
- 230000005750 disease progression Effects 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 238000007908 dry granulation Methods 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 230000006571 energy metabolism pathway Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 230000003090 exacerbative effect Effects 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000194 fatty acid Substances 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- 239000007888 film coating Substances 0.000 description 1
- 238000009501 film coating Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000037406 food intake Effects 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 229960002737 fructose Drugs 0.000 description 1
- 125000002446 fucosyl group Chemical group C1([C@@H](O)[C@H](O)[C@H](O)[C@@H](O1)C)* 0.000 description 1
- 230000003862 health status Effects 0.000 description 1
- 210000002216 heart Anatomy 0.000 description 1
- 238000001052 heteronuclear multiple bond coherence spectrum Methods 0.000 description 1
- 238000000990 heteronuclear single quantum coherence spectrum Methods 0.000 description 1
- 238000012165 high-throughput sequencing Methods 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 229940127130 immunocytokine Drugs 0.000 description 1
- 230000004957 immunoregulator effect Effects 0.000 description 1
- 229960003444 immunosuppressant agent Drugs 0.000 description 1
- 239000003018 immunosuppressive agent Substances 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 230000010226 intestinal metabolism Effects 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 150000002576 ketones Chemical class 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 239000008176 lyophilized powder Substances 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 210000004379 membrane Anatomy 0.000 description 1
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000003020 moisturizing effect Effects 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 210000004877 mucosa Anatomy 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- 238000000569 multi-angle light scattering Methods 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 238000000238 one-dimensional nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 239000000825 pharmaceutical preparation Substances 0.000 description 1
- 229940127557 pharmaceutical product Drugs 0.000 description 1
- 239000005015 poly(hydroxybutyrate) Substances 0.000 description 1
- 150000004804 polysaccharides Polymers 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 230000000529 probiotic effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000003805 procoagulant Substances 0.000 description 1
- 230000000770 proinflammatory effect Effects 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000012521 purified sample Substances 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
- 239000011541 reaction mixture Substances 0.000 description 1
- 230000001603 reducing effect Effects 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 150000004666 short chain fatty acids Chemical class 0.000 description 1
- 235000021391 short chain fatty acids Nutrition 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 238000004611 spectroscopical analysis Methods 0.000 description 1
- 238000001694 spray drying Methods 0.000 description 1
- 238000001370 static light scattering Methods 0.000 description 1
- 238000013179 statistical model Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 235000019605 sweet taste sensations Nutrition 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 230000004102 tricarboxylic acid cycle Effects 0.000 description 1
- 150000003628 tricarboxylic acids Chemical class 0.000 description 1
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 1
- WVLBCYQITXONBZ-UHFFFAOYSA-N trimethyl phosphate Chemical compound COP(=O)(OC)OC WVLBCYQITXONBZ-UHFFFAOYSA-N 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
- 235000019156 vitamin B Nutrition 0.000 description 1
- 239000011720 vitamin B Substances 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- UHVMMEOXYDMDKI-JKYCWFKZSA-L zinc;1-(5-cyanopyridin-2-yl)-3-[(1s,2s)-2-(6-fluoro-2-hydroxy-3-propanoylphenyl)cyclopropyl]urea;diacetate Chemical compound [Zn+2].CC([O-])=O.CC([O-])=O.CCC(=O)C1=CC=C(F)C([C@H]2[C@H](C2)NC(=O)NC=2N=CC(=CC=2)C#N)=C1O UHVMMEOXYDMDKI-JKYCWFKZSA-L 0.000 description 1
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明属于海洋技术领域，具体涉及一种具有抗结肠炎作用的海参多糖及其应用。本发明从玉足海参中分离纯化出一种全新结构的玉足海参精多糖H.leucospilota PS，由下氨基葡萄糖、葡萄糖醛酸、氨基半乳糖、半乳糖和岩藻糖5种单糖组成。该多糖通过改变肠道微生物群落结构，进一步的，调节3‑Hydroxybutanoate、L‑Malic acid、Succinic semialdehyde的产生，以此调节肠道能量代谢，改善结肠炎导致的肠道微生物群紊乱和能量代谢失调，能够避免长期服用结肠炎治剂所带来的耐药性和副作用。
The present invention belongs to the field of marine technology, and specifically relates to a sea cucumber polysaccharide with anti-colitis effect and its application. The present invention separates and purifies a sea cucumber polysaccharide H. leucospilota PS with a completely new structure from sea cucumbers, which is composed of five monosaccharides: glucosamine, glucuronic acid, aminogalactose, galactose and fucose. The polysaccharide changes the structure of intestinal microbial communities, and further regulates the production of 3-Hydroxybutanoate, L-Malic acid and Succinic semialdehyde, thereby regulating intestinal energy metabolism, improving intestinal microbial community disorders and energy metabolism disorders caused by colitis, and can avoid drug resistance and side effects caused by long-term use of colitis treatment agents.
Description
Technical Field
The invention belongs to the technical field of ocean, and particularly relates to sea cucumber polysaccharide with an anti-colonitis effect and application thereof.
Background
Ulcerative Colitis (UC) and Crohn's Disease (CD) are two major forms of Inflammatory Bowel Disease (IBD). The pathogenesis of UC is closely related to immune response disorders, changes in intestinal microbiota, genetic susceptibility and environmental factors. UC patients are often accompanied by reduced diversity of intestinal microbiota, altered metabolic profiles of intestinal microbiota, and reduced short chain fatty acids. Currently, UC is mainly treated by drugs, and clinicians can purposefully select drugs to control disease progression according to the severity of lesions in UC patients. Conventional formulations for UC, including aminosalicylic acid, corticosteroids, immunosuppressants and biologicals, however, these drugs have side effects and limitations. Therefore, there is a need to find new, safe and efficient natural products to prevent and improve UC.
Clinical evidence suggests that a decrease in fecal microbiota diversity is a common indicator of UC. When inflammation occurs in the intestinal tract, the number of intestinal flora is drastically reduced, the flora structure is destroyed, and some harmful bacteria are dominant, further exacerbating the inflammation. There is a correlation between intestinal flora characteristics and metabolite composition in UC patients. The relative abundance of certain pathogenic bacteria in UC is significantly correlated with the level of host microbial co-metabolites, while biogenic amines, amino acids and lipids are significantly increased and B vitamins are significantly reduced when the relative abundance of pathogenic bacteria is significantly increased. Intestinal metabolic biomarkers may improve methods of UC diagnosis and management, thereby facilitating the development and application of personalized medicine.
In recent years, researchers have found that a large number of functional polysaccharides can be obtained from many plants, animals and microorganisms in the ocean, and that these natural polysaccharides have advantages of safety, high therapeutic effect, non toxicity, low cost, good biocompatibility, and the like. The marine dietary polysaccharide has good immunoregulatory activity and anti-inflammatory effect, and influences the immune system through the regulation of intestinal flora. Chinese patent application CN111543638B discloses a sea cucumber polysaccharide product with auxiliary blood sugar reducing effect and a preparation method thereof, and the sea cucumber polysaccharide product reduces blood sugar of diabetics. Patent application CN111265627B discloses a sea cucumber polysaccharide composition for regulating intestinal flora, a preparation method and application thereof, and the sea cucumber polysaccharide composition can effectively regulate the intestinal flora, protect gastrointestinal mucosa and further improve immunity. Patent CN111658664B discloses application of sea cucumber polysaccharide in resisting novel coronavirus, and the sea cucumber polysaccharide oral liquid can inhibit invasion of SARS-CoV-2 virus from digestive system. However, the effect and application of Holothuria nobilis selenka polysaccharide H.leucospilota PS obtained by a specific method in the aspect of resisting colon inflammation have not been reported.
Disclosure of Invention
Aiming at the defects of the technology of the existing sea cucumber polysaccharide as a medicinal preparation in the anti-colonitis field, the invention aims to provide Holothuria nobilis selenka refined polysaccharide H.leucospilota PS with remarkable anti-colonitis activity and application thereof.
The object of the first aspect of the invention is to provide a sea cucumber polysaccharide.
The object of the second aspect of the present invention is to provide a method for preparing sea cucumber polysaccharide according to the first aspect of the present invention.
The object of the third aspect of the present invention is to provide a polysaccharide composition.
The fourth aspect of the invention aims at providing the application of the sea cucumber polysaccharide in the first aspect in preparing products for treating diabetic nephropathy.
In order to achieve the above purpose of the present invention, the present invention adopts the following technical scheme:
The object of the first aspect of the present invention is to provide a sea cucumber polysaccharide isolated from Holothuria nobilis, named Holothuria nobilis refined polysaccharide (Holothuria leucospilotapolysaccharide, H.leucopiplata PS).
Preferably, the sea cucumber polysaccharide comprises a polysaccharide represented by the structure of formula 1):
Formula 1);
wherein, the R group is selected from one of H, SO 3 -;
the R1 group is selected from one of Fucp, H, SO 3 -;
n is 75-157, p is pyranose (pyranose) abbreviation, representing a six-membered sugar.
In some embodiments of the invention, the sea cucumber polysaccharide consists of D-glucuronic acid (Glcp UA), N-acetylgalactosamine (GalpNAc) and fucose (Fucp), wherein the molar ratio of the D-glucuronic acid (Glcp UA), the N-acetylgalactosamine (GalpNAc) to the fucose (Fucp) is (0.2-0.6): 1 (2.0-3.0).
In some embodiments of the present invention, the sea cucumber polysaccharide is a sulfated polysaccharide having a Fucosylated Glycosaminoglycan (FGs) -like structure, and the main chain structure thereof is → 3) - β -D-Galp NAc- (1 → 4) - β -D-Glcp UA- (1 → where p is pyranose (pyranose) abbreviation, representing a hexahydric saccharide.
In some embodiments of the invention, the sea cucumber polysaccharide has a substitution group wherein the R group substitution occurs at the 4-O and 6-O positions of GalpNAc and the R group is selected from one of H, SO 3 -. The Fucp branch is located at the C3 site of GlcpUA, wherein the 2C and 3C sites of Fucp have an alternative R group, the R group being selected from one of H, SO 3 -, the 4C site of Fucp having an alternative R 1 group, the R 1 group being selected from one of Fucp, H, SO 3 -.
In some embodiments of the invention, the sea cucumber polysaccharide comprises a polysaccharide represented by the structure of formula 2):
Formula 2).
In some embodiments of the invention, the sea cucumber polysaccharide comprises a polysaccharide represented by the structure of formula 3):
Formula 3).
In some embodiments of the invention, the sea cucumber polysaccharide comprises a polysaccharide represented by the structure of formula 4):
formula 4).
In some embodiments of the invention, the polysaccharide of formula 1) has a weight average molecular weight of 9X 10 4~1.1×105 Da, preferably 1.05X10 5~1.08×105 Da.
In some embodiments of the invention, the polysaccharide of formula 1) has a number average molecular weight of 0.5X10 5~0.8×105 Da, preferably 0.6X10 5~0.7×105 Da.
In some embodiments of the invention, the polysaccharide of formula 1) has a Z-average molecular weight of 5X 10 5~7×105 Da, preferably 6X 10 5~6.5×105 Da.
In a second aspect of the present invention, there is provided a method for preparing sea cucumber polysaccharide according to the first aspect of the present invention, comprising the steps of:
1) Degreasing sea cucumber, and performing enzymolysis;
2) Precipitation of cetylpyridinium chloride (CPC);
3) Alcohol precipitation;
4) Decolorizing with active carbon;
5) Deproteinization;
6) Dialyzing for desalting;
7) And (5) chromatographic purification.
In some embodiments of the invention, the degreasing is performed with an organic reagent comprising at least one of acetone, petroleum ether, chloroform.
In some embodiments of the invention, the enzyme that is taken by the enzymolysis comprises at least one of papain, alkaline protease, trypsin.
In some embodiments of the invention, the temperature of the enzymatic hydrolysis is 50-70 ℃, the time of the enzymatic hydrolysis is 22-26 hours, and the enzyme dosage in the enzymatic hydrolysis is 8000-11000 (U/g).
In some embodiments of the invention, the CPC precipitate is used in an amount of 0.08-0.12 g/mL.
In some embodiments of the invention, the reagent used for the alcohol precipitation is selected from at least one of ethanol, isopropanol, butanol.
In some embodiments of the invention, the deproteinization method is selected from the group consisting of Sevage reagent or trichloroacetic acid.
In some embodiments of the invention, the chromatographic purification comprises ion chromatographic purification and molecular sieve chromatographic purification.
In some embodiments of the present invention, the ion chromatography purification is performed by DEAE Sepharose fast flow chromatography columns, and other conventional ion chromatography columns in the art can achieve corresponding effects.
In some embodiments of the invention, the molecular sieve chromatographic purification is performed by using a Sephadex G-100 chromatographic column, and other conventional molecular sieve chromatographic columns in the field can achieve corresponding effects.
In a third aspect, the invention provides a sea cucumber polysaccharide composition, which comprises Holothuria nobilis selenka polysaccharide H.leucospilota and pharmaceutically acceptable auxiliary materials.
In some embodiments of the invention, the pharmaceutically acceptable excipients include at least one of flavoring agents, binders, fillers.
In some embodiments of the invention, the flavoring agent is selected from at least one of xylitol, dried longan powder, sorbitol, maple syrup powder, and candy.
In some embodiments of the invention, the binder comprises at least one of chitosan, trehalose, acacia.
In some embodiments of the invention, the filler comprises at least one of corn starch, acetylated starch, hydroxypropyl starch.
In some embodiments of the invention, the mass ratio of the Holothuria nobilis selenka polysaccharide H.leucospilota, the flavoring agent, the adhesive and the filler powder is 6-12:15-20:35-45:10-15.
In a fourth aspect, the invention provides an application of the sea cucumber polysaccharide in the first aspect in preparing a product for treating colonitis.
In some embodiments of the invention, the holothuria nobilis selenka refined polysaccharide H.leucospilota PS relieves colon inflammation by modulating intestinal bacterial disorders and intestinal metabolites caused by colitis.
In some embodiments of the invention, the intestinal flora is Lactobacillaceae.
In some embodiments of the invention, the intestinal flora is Bifidobacteriaceac.
In some embodiments of the invention, the intestinal metabolite is 3-Hydroxybutanoate.
In some embodiments of the invention, the intestinal metabolite is L-MALIC ACID.
In some embodiments of the invention, the intestinal metabolite is Succinic semialdehyde.
In some embodiments of the invention, the product comprises a pharmaceutical product.
In some embodiments of the invention, the dosage form of the food or drug comprises at least one of powder, tablet, capsule, sustained release agent, granule.
The beneficial effects of the invention are as follows:
The invention separates and purifies holothuria leucospilota refined polysaccharide H.leucospilota PS with a brand new structure from holothuria leucospilota, and the holothuria leucospilota refined polysaccharide H.leucospilota is composed of 5 monosaccharides including glucosamine, glucuronic acid, galactosamine, galactose and fucose. The Holothuria nobilis selenka polysaccharide H.leucospilota essence polysaccharide PS further regulates the generation of 3-Hydroxybutanoate, L-MALIC ACID and Succinic semialdehyde by changing the intestinal microbiota structure, so as to regulate intestinal energy metabolism, improve intestinal microbiota disorder and energy metabolism disorder caused by colonitis, and avoid drug resistance and side effects caused by long-term administration of colonitis treatment agents.
2) The Holothuria nobilis selenka refined polysaccharide H.leucospilota holothurian extract PS obtained through separation and purification can better improve intestinal energy metabolism disorder caused by colonitis, so that the polysaccharide has a certain application prospect in preparation of medicines for treating colonitis.
Drawings
The invention is further described with reference to the accompanying drawings and examples, in which:
FIG. 1 shows HPAEC-PAD chromatograms of standard monosaccharides and H.leucopilostaPS (A) and high performance liquid chromatograms of H.leucopilostaPS (B).
Fig. 2 is an HPSEC chromatogram of h.
Fig. 3 is an infrared spectrum of h.
FIG. 4 is a 1H NMR spectrum of H.leucopirotaPS.
FIG. 5 is a 13C NMR spectrum of H.leucopirotaPS.
FIG. 6 is a COSY spectrum of H.leucopirotaPS.
Fig. 7 is a TOCSY spectrum of h.
FIG. 8 is the HSQC spectrum of H.leucopirotaPS.
FIG. 9 is a NOESY spectrum of H.leucopirotaPS.
Fig. 10 is an HMBC spectrum of h.
Fig. 11 is a schematic structural diagram of h.
FIG. 12 shows the results of the changes in the amounts of immunocytokines TNF-alpha and IL-6 in the serum of example 3.
FIG. 13 shows the results of the relative content change of microorganisms Lactobacillaceae, bifidobacteriaceac in the intestinal tract.
FIG. 14 shows the results of the relative content changes of 3-Hydroxybutanoate, L-MALIC ACID, succinic semialdehyde in feces.
Detailed Description
The conception and the technical effects produced by the present invention will be clearly and completely described in conjunction with the embodiments below to fully understand the objects, features and effects of the present invention. It is apparent that the described embodiments are only some embodiments of the present invention, but not all embodiments, and that other embodiments obtained by those skilled in the art without inventive effort are within the scope of the present invention based on the embodiments of the present invention.
EXAMPLE 1 separation and purification of Holothuria nobilis selenka refined polysaccharide H.leucospilota PS
According to the extraction method of the sea cucumber body wall polysaccharide, holothuria nobilis is selected as the sea cucumber, and Holothuria nobilis is one of the most common edible sea cucumbers in south China sea, and has medicinal and edible values.
The method comprises the following steps of soaking the dried sea cucumber body wall for 20-24 hours, removing viscera and body cavity membranes, mincing by a meat mincer, and freeze-drying to obtain powder, wherein the freeze-drying parameters of the freeze-dried powder are-60 to-25 ℃, and the vacuum degree is 1-3 bar and 10-14 h. Soaking in acetone for degreasing, wherein the mass ratio of sodium acetate to papain to EDTA to cysteine to water is 3.28:0.26:0.58:0.24. Stirring and centrifuging, collecting supernatant, adding cetylpyridinium chloride (Cetyl-Pyridinium Chloride, CPC) into the supernatant at a ratio of 10% (W/V), standing at room temperature, centrifuging, and redissolving the precipitate in NaCl-ethanol (100:15V/V) solution. And adding absolute ethyl alcohol for precipitation, standing for 10-12h, and centrifuging. Adding 0.5% -0.75% active carbon into the supernatant for decoloring, and stirring for decoloring at 40 ℃ for 60 min. Trifluoroacetic acid is used for removing protein. Dialyzing with dialysis bag with molecular weight cutoff of 8000 Da for 48-h, and lyophilizing (at-60deg.C to-25deg.C and vacuum degree of 1-3 bar) for 10-14-h to obtain sea cucumber crude polysaccharide.
The invention relates to a fractional column chromatography, which is characterized in that DEAE-Sepharose Fast Flow column is adopted for fractional separation, and after elution, sephadex G-100 column is used for purification. After the column was assembled, the well degassed DEAE-FF packing was packed and equilibrated with 0.1M sodium acetate buffer. 200 mg samples were weighed, dissolved in 10mL buffer, and the prepared polysaccharide solution was filtered through a 0.45 μm filter and then added to a chromatographic column. The eluent was treated with a sodium acetate solution of 0.1M containing 1.5M, a flow rate of 1 mL/min was set, 5 min/tube, and 61-68 th tube was collected and lyophilized to give HLP-1. Further purification using Sephadex G-100 by weighing 100 mg of HLP-1 lyophilized powder, adding to 5mL of 0.1M sodium acetate buffer containing 2M NaCl, and passing through 0.45 μm filter membrane after sufficient dissolution, the flow rate was set at 0.5 mL/min. The main peak was collected (tubes 19-30), 5mL per tube. Concentrating under reduced pressure, dialyzing 48 h, and lyophilizing to obtain purified sample Holothuria nobilis selenka refined polysaccharide.
Example 2 structural identification of Holothuria nobilis selenka refined polysaccharide H.leucospilota PS
1) High performance liquid chromatography monosaccharide composition analysis
Monosaccharide composition was measured by High Performance Liquid Chromatography (HPLC) method of pre-column PMP derivatization. A sample of 6 mg (prepared in example 1) was dissolved with 3 mL trifluoroacetic acid (4M) to prepare a 2 mg/mL mixture, which was transferred to an ampoule for sealing and hydrolyzed at 120℃to 4 h. After the reaction was completed, the reaction mixture was cooled to room temperature, 100. Mu.L of the hydrolysate was added with an equal volume of methanol, and the mixture was dried with nitrogen to remove trifluoroacetic acid, and the above-mentioned operation was repeated 3 times until the weight of the sample tube was constant. The dried sample was dissolved in 100. Mu.L of ultra pure water, and after adding an equal volume of 0.6M NaOH solution and mixing well, the pre-column derivatization reaction was carried out by continuing to add an equal volume of 0.5M PMP-methanol solution, and the reaction was carried out in a 70℃water bath for 1.5 h. After the reaction was cooled to room temperature, 100. Mu.L of the reaction product was taken out and neutralized with 50. Mu.L of hydrochloric acid (0.3M), chloroform 1mL and 900. Mu.L of water were added, mixed with vigorous shaking and centrifuged 5min, the chloroform layer was discarded, and the aqueous layer was further added with 1mL chloroform, and extraction was repeated 3 times. Finally, the upper layer (water phase layer) is filtered by a water-based filter membrane with the thickness of 0.22 mu m, and then high performance liquid chromatography detection is carried out. The monosaccharide standard is prepared into a solution with the concentration of 1 mg/mL and the polysaccharide hydrolysate is subjected to derivatization labeling simultaneously. The monosaccharide standard is glucosamine, glucuronic acid, galactosamine, galactose and fucose.
The detection conditions of the liquid chromatography are shown in table 1.
TABLE 1 liquid chromatography operating parameters
2) Analysis of monosaccharide composition by ion chromatography
And (3) passing the upper layer (aqueous layer) derivatization polysaccharide solution obtained in the step (1) through a 0.22 mu m aqueous filter membrane, and injecting samples. The monosaccharide standard solution and the mixed standard solution with different concentrations are prepared, and the sample is analyzed by using a HPACE-PAD method. The mixed standard solution is prepared from glucosamine, glucuronic acid, galactosamine, galactose and fucose with the same concentration, 5 monosaccharide standard samples are respectively prepared into stock solutions with the concentration of 5.0mg/mL, 1.0mL of each stock solution of the 5 monosaccharide standard samples is absorbed, the stock solutions are mixed to prepare mixed standard solutions with the concentration of 0.5mg/mL, and the mixed standard solutions are diluted to the required concentration gradient, such as the mixed standard solutions diluted to 0.02, 0.04, 0.06, 0.08 and 0.1 mg/mL. A carbon Pac TMPA 20 (4 mm X250 mm) ion chromatography column was fitted, a flow rate of 0.5 mL/min was set, mobile phase A was UP water, expressed as a percentage.
TABLE 2 monosaccharide gradient elution procedure
Due to limitations of High Performance Anion Exchange Chromatography (HPAEC) and PA-20 column separation effect, peak times of galactosamine and rhamnose are very close (a in fig. 1). In structural analysis, HPSEC and PMP-HPLC were again used with Agilent C18 column to measure monosaccharide composition. The peaks were quantified by integration according to the position and time of the HPLC peak (B in FIG. 1), and the molar ratio of the monosaccharide composition of HLP-I, mainly D-glucuronic acid (Glcp UA), N-acetylgalactosamine (GalpNAc) and fucose (Fucp), was found to be 0.45:1.00:2.40.
3) Determination of homogeneity and molecular weight
High performance size exclusion chromatography-multi angle laser light scattering (HPSEC-MALLS) was measured using an Agilent 1200 series LC system (Agilent Technologies, palo Alto, USA) at 25 ℃. The mobile phase was a 0.1M NaNO 3 aqueous solution, the flow rate was 0.6 mL/min, and the specific refractive index increment (dn/dc) was set to 0.145 mL/g. The molecular weight is calculated by the Zimm method of static light scattering according to the basic light scattering equation.
The weight average molecular weight (Mw), number average molecular weight (Mn) and Z average molecular weight (Mz) of the H.leucopiplatPS were determined to be 1.062X 10 5Da、0.652×105 Da and 6.232X 10 5 Da, respectively. The narrow and symmetrical peaks on the HPSEC-RI chromatogram (fig. 2) and the relatively low values of polydispersity index (1.63, pdi=mw/Mn) indicate higher purity and better homogeneity of h.leucopitata ps.
4) Molecular structure analysis by infrared spectrum
The sample and KBr mixture (1:100, w/w) was compressed into flakes and transferred to a FT-IR spectrometer (Nicolet 6700, thermo FISHER SCIENTIFIC Technology Co., ltd, USA). FT-IR spectra were recorded in the range 400-4000 cm -1 and scanned 64 times.
5) Nuclear magnetic resonance spectrum analysis molecular structure
Deuterated polysaccharide (50 mg) was dissolved in 0.6 mL D2o and 1D and 2D NMR experiments were performed. All experiments were performed on a Bruker AVANCE spectrometer (Bruker Biospin, rheinstetten, germany). 1H and 13C NMR spectra were acquired under 298.2K conditions using 600.58 and 151.01 MHz proton frequencies and were processed with MestReNova 14.0 software. Two-dimensional nuclear magnetic resonance experiments, including 1H-1H correlation spectroscopy (1H-1H COSY), 1H-13C Heteronuclear Single Quantum Coherence (HSQC), heteronuclear Multiple Bond Correlation (HMBC), total correlation spectroscopy (TOCSY) and effect spectroscopy (NOESY), were performed for structural analysis.
As shown in FIG. 3, the strong absorption band at 3478.53 cm -1 represents the stretching vibration of O-H in the sugar residue. 2929.85 The band around cm -1 is related to the stretching vibration of C-H in the sugar ring. Peaks at about 1739 cm -1 and 1650 cm -1 are attributed to the-COOH bending vibrations of the carboxylic acid product and to the bending and stretching vibrations of C-O in glucuronic acid and N-acetamido galactose. 1253.12 The signal at cm -1 is attributed to the asymmetric stretching vibration of the sulfate group (o=s=o), which is a typical feature of sulfated polysaccharides. 1050.93 The absorption peak of cm -1 is in the fingerprint region of the polysaccharide (400-1300: 1300 cm -1, which can be used to distinguish minor differences in structure from different compounds), and can identify specific chemical groups in HLP-I, which are generated by C-O-C stretching vibration. The asymmetric absorption peaks 962.47, 850.71 and 686.36 cm -1 also indicate the presence of abundant sulfate groups in h.leucopicitaps. 820 The band around cm -1- 850 cm-1 has a distinct difference, possibly related to the different substitution positions of the sulfuric acid groups in the polysaccharide chain structure. The characteristic peak of C-O-S stretching vibration at 850.71 cm -1 indicates that the C-4 position of the fucose residue contains an axial sulfate group.
As shown in FIG. 4, the chemical shift of the anomeric proton signal region in the 1H NMR spectrum was 5.0-5.8 ppm, indicating the presence of an alpha-type fucose sulfate residue and the type of sulfate group substitution in H.leucopitaPS was Fucp2,4S,Fucp4S,Fucp0S. The signal range of Fucp and GalNpAc for the methyl proton (-CH 3) is 1.1-1.4, ppm and 1.8-2.1, ppm, and signal integration is performed to obtain an area ratio of 1.05:1.00. The 13C NMR spectrum further illustrates that the carbon skeleton of H.leucopiplatPS polysaccharide is composed of L-Fucp and D-GalNpAc and D-GlcUA, and D-GalpNAc and D-GlcUA are in the beta anomeric configuration (FIG. 5).
From the HSQC spectral data of H.leucopitaPS (FIG. 6), the positions of the H-1 signals belonging to the H and I units were 5.62 ppm and 5.32 ppm, respectively. The 1H-1H COSY (FIG. 7) spectra were used to further analyze other signals of the resonance system. The presence of sulfate groups at both the C-2 and C-4 positions in the H unit was verified by the H-2 (4.39 ppm) and H-4 (4.58 ppm) signals at the low field positions. The H-3 (3.91 ppm) and H-4 (4.14 ppm) signals confirm the sulfation of the I units at O-3, O-4. As shown in FIG. 8, signals A, B, C are derived from (H-1, C-1), (H-2, C-2), (H-3, C-3), (H-4, C-4), (H-5, C-5) and (H-6, C-6) of GalpNAc, galpNAc S6S, galpNAc S. Signals D, E and F are derived from (H-1, C-1), (H-2, C-2), (H-3, C-3), (H-4, C-4) and (H-5, C-5) of Glcp UA, glcp UA2S and GlcpUA S3S. The chemical shifts of H-4 and H-6 in GalpNAc4S6S are at a lower field than GalpNAc, indicating that the O-4 and O-6 positions are sulfated. The residue sequence of the glycosidic bond was determined from the correlation peaks in HMBC and NOESY. The two distinct signals of 4.39/3.86 ppm and 4.64/4.43 ppm in FIG. 9 (designated D1/A3, B3, C3 and H1, B1, C1/D1, E1, F1, respectively) indicate that Glcp UA is linked to the C-3 position of GalN, which is associated with the C-3 position of GalpNAc. The cross peaks of the H1 (D)/H3 (E), H1 (G, I)/H3 (E) and H1 (G, I)/H3 (F) interactions in fig. 10 verify that Glcp UA has a fucosyl residue substitution at O-3.
H/C chemical shift data for leucopiclatPS, as shown in Table 3. Based on the structural analysis described above, h.leucopicitaps contains three main monosaccharides (GlcpUA, galpNAc and Fucp), the backbone structure is similar to some sea cucumber body wall FGs. Thus, it is speculated that H.leucopilotaPS is a sulfated glycosaminoglycan having a FGs-like structure, and that sulfuric acid substitution occurs at the O-4 and O-6 positions of GalpNAc. Furthermore, h.leucopiplata ps has fucose branches with different sulfation patterns (Fucp 2, 4S, fucp 3, 4S and Fucp S).
Assignment of 1 H and 13 C NMR spectra of Table 3 H.leucospilotaPS
Based on the above data, h.leucopiplata includes the following three different polysaccharide formulas:
formula 2);
Formula 3);
formula 4).
The structural schematic of the predicted h.leucopitata ps is shown in fig. 11, in combination with infrared and nuclear magnetic data.
Example 3 H.leucospilotaPS therapeutic Effect on inflammatory bowel disease
Animal experiment design the study was approved by the university of hainan animal ethical committee and all animal procedures were performed according to hainan university, guidelines for laboratory animal care and use. After the male SD rats were adaptively cultured in the experimental environment for 7 days, the SD rats were randomly divided into 6 groups, which were respectively a normal control group (NC, lavage saline, drinking purified water), a model control group (MC, lavage saline, drinking DSS aqueous solution), a positive control group (PC, lavage 100 mg/kg mesalazine, drinking DSS aqueous solution), a H.leucopirotaPS low dose group (HLP-L, lavage 50 mg/kg HLP, drinking DSS aqueous solution), a H.leucopirotaPS medium dose group (HLP-M, lavage 100 mg/kg HLP, drinking DSS aqueous solution), a H.leucopirotaPS high dose group (HLP-H, lavage 200 mg/kg HLP, drinking DSS aqueous solution). HLP group rats were gavaged for the first week of experiment, each group had free access to water for ingestion. The second week of the experiment, except for NC group, 3% dss (36,000-50,000 mw) was added to the drinking water of the remaining groups, and ulcerative colitis was induced by continuous drinking for 7 days. At the same time, rats in PC group and HLP group (HLP-L group, HLP-M group and HLP-H group) were respectively intragastric 5-ASA and different doses of HLP for intervention.
2) Sample collection and inflammatory factor determination
The health status of the experimental rats was checked daily for the second week of the experiment, and fresh fecal samples were collected the day before the end of the experiment and flash frozen in liquid nitrogen. After a fast of 12h, all experimental rats were anesthetized with 5% chloral hydrate (10 g/0.1 mL), collected from the orbit, and whole blood centrifuged (3500 g,10 min,4 ℃) to give serum, which was immediately placed in-80 ℃ for frozen storage for subsequent analysis. The cytokines IL-6 and TNF-alpha in rat serum were detected by enzyme-linked immunosorbent assay (ELISA) and the specific procedures were performed according to the instructions of the IL-6 kit (Jiangsu enzyme Biotechnology Co., ltd., cat# MM-0190R 1) and the TNF-alpha kit (Prriley APPLYGEN, cat# AZ 0672).
3) 16S rDNA high throughput sequencing analysis
After extracting the total genome DNA in the fecal sample, the mass of the total genome DNA is detected by 1% agarose gel electrophoresis and quantified by a spectrophotometer. And TRANSSTART FASTPFU DNA polymerase is adopted in PCR amplification, and meanwhile, the amplification cycle number is controlled, so that the amplification conditions of the same batch of samples are ensured to be consistent. After the amplification step is completed, the PCR product is recovered by cutting gel using a gel recovery kit. Elution and detection was performed by 2% agarose electrophoresis. And quantifying the PCR product by using a fluorescence quantification system according to the electrophoresis preliminary quantification result. The P7 linker sequence of Illumina (CAAGCAGAAGACGGCATACGAGAT, SEQ ID NO: 1) was added to the outer end of the target region by PCR, the PCR product was recovered by gel cutting using a gel recovery kit, and sodium hydroxide was denatured to generate single-stranded DNA fragments. Sequencing on a machine and analyzing the result.
4) Analysis of metabolites in feces
The rat fecal sample was removed from the ultra-low temperature freezer and thawed in a 4 ℃ freezer. 100mg of fecal sample was taken, 1200. Mu.L of methanol/acetonitrile/water extract (2:2:1), vortexed 2min, sonicated at room temperature for 10min, and allowed to stand in a 4℃refrigerator for 10 min. Centrifuge 15 min at 14,000 g at 4℃and prepare the supernatant for film coating and test on machine.
5) Experimental results
TNF- α is a potent cytokine in IBD that promotes inflammation through adhesion molecule expression, fibroblast proliferation, and procoagulant factor activation, among others. The increased expression of IL-6 is associated with an increased risk of colorectal adenoma development in experimental colitis and IBD patients, with high production of IL-6 by lamina propria macrophages and CD4 + T cells. The results are shown in FIG. 12, where DSS significantly increased the levels of the pro-inflammatory factors TNF- α, IL-6 in rat serum, and the levels of cytokines were recovered to different extents in the HLP and PC groups, as compared to the NC group. The intervention of H.leucopirotaPS is shown to significantly improve colon inflammation, has an inflammatory factor inhibition effect similar to that of 5-ASA, and has no toxic or side effect compared with that of 5-ASA taken for a long time.
The results of flora sequencing are shown in FIG. 13, and the relative abundance of Lactobacillaceae, bifidobacteriaceac in the intestinal flora of rats is down-regulated after DSS induction. The relative abundance of these flora was significantly improved by HLP treatment. Lactobacillaceae is the most abundant probiotic, and generates single-chain fatty acid by fermenting oligosaccharides and polysaccharides, so as to inhibit adhesion of pathogens and intestinal epithelial cells, improve the environment in the intestinal tract and promote proliferation of acidophilic probiotics. Lactobacillaceae, bifidobacteriaceac can improve intestinal environment, inhibit growth of harmful bacteria, and enhance intestinal immune barrier.
The results of fecal metabolites are shown in fig. 14, and according to the multiple of change >2 between groups, the occurrence probability of the significance result <0.05 and the predicted result of a single variable in a statistical model to the model >1.0, the differential metabolites between groups are screened, the metabolites 3-Hydroxybutanoate, L-MALIC ACID and Succinic semialdehyde with obvious differences are screened out from the fecal metabolites and used as the metabolites with obvious differences of colonitis intestinal metabolism, the contents of 3-Hydroxybutanoate and Succinic semialdehyde in colonitis rat feces are reduced, and the content of L-MALIC ACID is increased, so that DSS interferes with the intestinal energy metabolism of rats. After intake of HLP, the content of 3-Hydroxybutanoate, succinic semialdehyde in feces increases and the content of L-MALIC ACID decreases. Unlike other organ energy sources, intestinal energy is 70% from the oxidation of butyric acid. Thus, butyric acid is the primary energy source for intestinal epithelial cells. Inflammation in the colon is a complex energy consuming process, and when the content of substances involved in energy metabolism changes significantly, it is indicated that the colon is abnormal in energy metabolism. L-MALIC ACID and Succinic semialdehyde are intermediates of tricarboxylic acid cycles. 3-Hydroxybutanoate is a monomer of microbial polyhydroxybutyrate, and is also a natural ketone body generated by the oxidation of butyrate in gastrointestinal epithelial cells, and provides energy for cells, hearts and brains. Thus, HLP regulates intestinal flora and restores normal energy supply to intestinal epithelial cells (oxidative energy in butyrate), improving the intestinal environment, revealing the mechanism by which HLP improves colonic inflammation in rats by improving intestinal flora, regulating energy metabolic pathways (such as tricarboxylic acid circulation pathways).
In conclusion, the Holothuria nobilis selenka refined polysaccharide H.leucospilota PILOTAPS obtained through separation and purification can better improve intestinal energy metabolism disorder caused by colonitis, and the polysaccharide has a certain application prospect in preparing a medicament for treating colonitis.
Example 4A sea cucumber polysaccharide microgel enteric sustained-release tablet
An enteric-coated slow-release tablet of sea cucumber polysaccharide microcapsule is prepared from sea cucumber polysaccharide (10 g), chitosan (5 g) as embedding material for providing slow-release effect, trehalose (10 g) as plant-derived adhesive, natural sweetener with similar film-forming property, xylitol (10 g) as natural sweetener with moisture-retaining property, longan dry powder (30 g for increasing sweet taste and nutritive value) and hydroxypropyl starch (15 g) through wet granulating. The enteric coated tablet has good effect on improving colonitis.
Example 5A sea cucumber polysaccharide microgel enteric sustained-release tablet
The formulation of the sea cucumber polysaccharide microcapsule enteric sustained-release tablet comprises 8g of sea cucumber polysaccharide, 6g of chitosan (enhancing embedding and sustained-release effects), 10g of acacia (serving as a plant source, serving as a binder and a thickening agent), 10g of sorbitol (having moisturizing and sweetening effects), 30g of maple syrup powder (a natural source sweetener and good taste) and 10g of modified corn starch (having good fluidity and cohesiveness), the microcapsule sustained-release enteric-release granule is obtained by spray drying, and the sea cucumber polysaccharide microcapsule enteric-release tablet is obtained by granulating. The enteric coated tablet has good effect on improving colonitis.
Example 6A sea cucumber polysaccharide microgel enteric sustained-release tablet
The formula of the enteric-coated slow-release granule of the sea cucumber polysaccharide microcapsule comprises 7g of sea cucumber polysaccharide, 8g of chitosan (providing additional biocompatibility and intestinal protection), 10g of carrageenan, 10g of mannitol, 30g of preserved fruit sugar and 15g of acetylated starch, and the enteric-coated tablet of the microcapsule is obtained by dry granulation. The enteric coated tablet has good effect on improving colonitis.
Claims (10)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202411612593.7A CN119119321A (en) | 2024-11-13 | 2024-11-13 | Sea cucumber polysaccharide with anti-colitis effect and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202411612593.7A CN119119321A (en) | 2024-11-13 | 2024-11-13 | Sea cucumber polysaccharide with anti-colitis effect and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN119119321A true CN119119321A (en) | 2024-12-13 |
Family
ID=93752405
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202411612593.7A Pending CN119119321A (en) | 2024-11-13 | 2024-11-13 | Sea cucumber polysaccharide with anti-colitis effect and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN119119321A (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101863997A (en) * | 2010-05-24 | 2010-10-20 | 南京中医药大学 | Preparation method of sea cucumber polysaccharide |
-
2024
- 2024-11-13 CN CN202411612593.7A patent/CN119119321A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101863997A (en) * | 2010-05-24 | 2010-10-20 | 南京中医药大学 | Preparation method of sea cucumber polysaccharide |
Non-Patent Citations (1)
| Title |
|---|
| XIN ZHANG等: "Microbiome–Metabolomics Analysis Reveals the Mechanism of Holothuria leucospilota Polysaccharides(HLP) in Ulcerative Colitis", 《MOL. NUTR. FOOD RES.》, vol. 67, 22 March 2023 (2023-03-22), pages 1 - 11 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Shao et al. | Anti-inflammatory and intestinal microbiota modulation properties of Jinxiang garlic (Allium sativum L.) polysaccharides toward dextran sodium sulfate-induced colitis | |
| Zhang et al. | Anti-inflammatory activity of alkali-soluble polysaccharides from Arctium lappa L. and its effect on gut microbiota of mice with inflammation | |
| Zhang et al. | Catabolism of Saccharina japonica polysaccharides and oligosaccharides by human fecal microbiota | |
| Cai et al. | Structural characterization of oligosaccharide from Spirulina platensis and its effect on the faecal microbiota in vitro | |
| CN105848500B (en) | Nutritional composition with lipid metabolism improving effect | |
| KR20160137581A (en) | Nutritional composition | |
| Wu et al. | In vitro digestive characteristics and microbial degradation of polysaccharides from lotus leaves and related effects on the modulation of intestinal microbiota | |
| Wu et al. | RGI-type pectic polysaccharides modulate gut microbiota in a molecular weight-dependent manner in vitro | |
| WO2021196572A1 (en) | Fucose-rich extracellular polysaccharide, preparation method therefor and application thereof | |
| Chen et al. | Improving the digestive stability and prebiotic effect of carboxymethyl chitosan by grafting with gallic acid: In vitro gastrointestinal digestion and colonic fermentation evaluation | |
| Jing et al. | Structural characterization of a heteropolysaccharide from the fruit of Crataegus pinnatifida and its bioactivity on the gut microbiota of immunocompromised mice | |
| Lu et al. | Effects of exopolysaccharides from Antrodia cinnamomea on inflammation and intestinal microbiota disturbance induced by antibiotics in mice | |
| Sun et al. | Digestion characteristics of polysaccharides from Gracilaria lemaneiformis and its interaction with the human gut microbiota | |
| Pan et al. | A galacturonan from Dioscorea opposita Thunb. regulates fecal and impairs IL-1 and IL-6 expression in diarrhea mice | |
| Xu et al. | Studies of peach gum polysaccharide on gut microbiota in vitro fermentation by human feces | |
| Qin et al. | Alleviative effects of sulfated polysaccharide from Ishige Okamurae against DSS-induced ulcerative colitis via inhibiting inflammation and modulating gut microbiota | |
| Gu et al. | Metabolic outcomes of Cordyceps fungus and Goji plant polysaccharides during in vitro human fecal fermentation | |
| Zhang et al. | Extraction of polysaccharides from Polygonum cuspidatum with activity against Type 2 Diabetes via alterations in gut microbiota | |
| CN115260330A (en) | Inulin type fructan extracted from stevia rebaudiana Bertoni root, and its preparation method and application | |
| Wang et al. | Structure analysis and in vitro evaluation of probiotic properties for polysaccharides from Phellinus baumii extracted with phosphotungstic acid assistance | |
| Bai et al. | Purification, characterization, simulated gastrointestinal digestion and gut microbiota fermentation of a Bifidobacterium-directed mannoglucan from Lilium brownii var. viridulum | |
| Zhou et al. | In vivo absorption and fecal excretion of polysaccharides from the fruits of Lycium barbarum L. in rats through fluorescence labeling | |
| Jing et al. | Structural characterization of polysaccharide from the peel of Trichosanthes kirilowii Maxim and its anti-hyperlipidemia activity by regulating gut microbiota and inhibiting cholesterol absorption | |
| Tang et al. | Advanced structural characterization and in vitro fermentation prebiotic properties of cell wall polysaccharide from Kluyveromyces marxianus | |
| CN116284468B (en) | Tremella aurantialba acidic polysaccharide, preparation method thereof and application thereof in improving ulcerative colitis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |