CN119097046A - An animal feed additive with anti-inflammatory and antioxidant functions - Google Patents
An animal feed additive with anti-inflammatory and antioxidant functions Download PDFInfo
- Publication number
- CN119097046A CN119097046A CN202411224968.2A CN202411224968A CN119097046A CN 119097046 A CN119097046 A CN 119097046A CN 202411224968 A CN202411224968 A CN 202411224968A CN 119097046 A CN119097046 A CN 119097046A
- Authority
- CN
- China
- Prior art keywords
- animal feed
- feed additive
- subcritical
- petroleum ether
- acid
- 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
- 235000019730 animal feed additive Nutrition 0.000 title claims abstract description 22
- 230000003110 anti-inflammatory effect Effects 0.000 title abstract description 16
- 230000003078 antioxidant effect Effects 0.000 title abstract description 9
- 239000003963 antioxidant agent Substances 0.000 title abstract description 6
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims abstract description 28
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 20
- 229920001184 polypeptide Polymers 0.000 claims abstract description 19
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 19
- 238000000605 extraction Methods 0.000 claims abstract description 15
- 241001465754 Metazoa Species 0.000 claims abstract description 14
- 239000003208 petroleum Substances 0.000 claims abstract description 14
- 239000000843 powder Substances 0.000 claims abstract description 13
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 10
- 240000007817 Olea europaea Species 0.000 claims abstract description 10
- 238000004821 distillation Methods 0.000 claims abstract description 5
- 238000002386 leaching Methods 0.000 claims abstract description 4
- 238000002360 preparation method Methods 0.000 claims abstract description 3
- 238000002156 mixing Methods 0.000 claims description 11
- 208000004232 Enteritis Diseases 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- 235000013399 edible fruits Nutrition 0.000 claims description 6
- LCGLNKUTAGEVQW-UHFFFAOYSA-N Dimethyl ether Chemical compound COC LCGLNKUTAGEVQW-UHFFFAOYSA-N 0.000 claims description 4
- ATUOYWHBWRKTHZ-UHFFFAOYSA-N Propane Chemical compound CCC ATUOYWHBWRKTHZ-UHFFFAOYSA-N 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 4
- NNPPMTNAJDCUHE-UHFFFAOYSA-N isobutane Chemical compound CC(C)C NNPPMTNAJDCUHE-UHFFFAOYSA-N 0.000 claims description 4
- 239000000463 material Substances 0.000 claims description 4
- 239000000706 filtrate Substances 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 239000002245 particle Substances 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 239000001273 butane Substances 0.000 claims description 2
- 239000001282 iso-butane Substances 0.000 claims description 2
- IJDNQMDRQITEOD-UHFFFAOYSA-N n-butane Chemical compound CCCC IJDNQMDRQITEOD-UHFFFAOYSA-N 0.000 claims description 2
- OFBQJSOFQDEBGM-UHFFFAOYSA-N n-pentane Natural products CCCCC OFBQJSOFQDEBGM-UHFFFAOYSA-N 0.000 claims description 2
- 239000001294 propane Substances 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 2
- MIJYXULNPSFWEK-GTOFXWBISA-N 3beta-hydroxyolean-12-en-28-oic acid Chemical compound C1C[C@H](O)C(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CCC(C)(C)C[C@H]5C4=CC[C@@H]3[C@]21C MIJYXULNPSFWEK-GTOFXWBISA-N 0.000 abstract description 39
- JKLISIRFYWXLQG-UHFFFAOYSA-N Epioleonolsaeure Natural products C1CC(O)C(C)(C)C2CCC3(C)C4(C)CCC5(C(O)=O)CCC(C)(C)CC5C4CCC3C21C JKLISIRFYWXLQG-UHFFFAOYSA-N 0.000 abstract description 39
- YBRJHZPWOMJYKQ-UHFFFAOYSA-N Oleanolic acid Natural products CC1(C)CC2C3=CCC4C5(C)CCC(O)C(C)(C)C5CCC4(C)C3(C)CCC2(C1)C(=O)O YBRJHZPWOMJYKQ-UHFFFAOYSA-N 0.000 abstract description 39
- MIJYXULNPSFWEK-UHFFFAOYSA-N Oleanolinsaeure Natural products C1CC(O)C(C)(C)C2CCC3(C)C4(C)CCC5(C(O)=O)CCC(C)(C)CC5C4=CCC3C21C MIJYXULNPSFWEK-UHFFFAOYSA-N 0.000 abstract description 39
- 229940100243 oleanolic acid Drugs 0.000 abstract description 39
- HZLWUYJLOIAQFC-UHFFFAOYSA-N prosapogenin PS-A Natural products C12CC(C)(C)CCC2(C(O)=O)CCC(C2(CCC3C4(C)C)C)(C)C1=CCC2C3(C)CCC4OC1OCC(O)C(O)C1O HZLWUYJLOIAQFC-UHFFFAOYSA-N 0.000 abstract description 39
- MDZKJHQSJHYOHJ-UHFFFAOYSA-N crataegolic acid Natural products C1C(O)C(O)C(C)(C)C2CCC3(C)C4(C)CCC5(C(O)=O)CCC(C)(C)CC5C4=CCC3C21C MDZKJHQSJHYOHJ-UHFFFAOYSA-N 0.000 abstract description 37
- MDZKJHQSJHYOHJ-LLICELPBSA-N maslinic acid Chemical compound C1[C@@H](O)[C@H](O)C(C)(C)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CCC(C)(C)C[C@H]5C4=CC[C@@H]3[C@]21C MDZKJHQSJHYOHJ-LLICELPBSA-N 0.000 abstract description 37
- 230000000694 effects Effects 0.000 abstract description 20
- 239000003674 animal food additive Substances 0.000 abstract description 10
- 230000007413 intestinal health Effects 0.000 abstract description 5
- 239000003814 drug Substances 0.000 abstract description 3
- 238000002474 experimental method Methods 0.000 abstract description 2
- 239000010465 pomace olive oil Substances 0.000 abstract 1
- 241000699670 Mus sp. Species 0.000 description 26
- 239000000243 solution Substances 0.000 description 19
- 238000012360 testing method Methods 0.000 description 14
- 238000002835 absorbance Methods 0.000 description 12
- 210000001072 colon Anatomy 0.000 description 12
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 9
- 239000003242 anti bacterial agent Substances 0.000 description 9
- 229940088710 antibiotic agent Drugs 0.000 description 9
- 238000000034 method Methods 0.000 description 9
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 8
- 241000699666 Mus <mouse, genus> Species 0.000 description 8
- 239000000523 sample Substances 0.000 description 8
- 210000001519 tissue Anatomy 0.000 description 8
- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 description 7
- 108010005636 polypeptide C Proteins 0.000 description 7
- 210000002966 serum Anatomy 0.000 description 7
- 102000004388 Interleukin-4 Human genes 0.000 description 6
- 108090000978 Interleukin-4 Proteins 0.000 description 6
- 229940028885 interleukin-4 Drugs 0.000 description 6
- 239000012488 sample solution Substances 0.000 description 6
- 102000016938 Catalase Human genes 0.000 description 5
- 108010053835 Catalase Proteins 0.000 description 5
- 108010063907 Glutathione Reductase Proteins 0.000 description 5
- 102100036442 Glutathione reductase, mitochondrial Human genes 0.000 description 5
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 description 5
- 102000003896 Myeloperoxidases Human genes 0.000 description 5
- 108090000235 Myeloperoxidases Proteins 0.000 description 5
- 102000019197 Superoxide Dismutase Human genes 0.000 description 5
- 108010012715 Superoxide dismutase Proteins 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 5
- 230000012010 growth Effects 0.000 description 5
- 229940118019 malondialdehyde Drugs 0.000 description 5
- 238000007254 oxidation reaction Methods 0.000 description 5
- 230000036542 oxidative stress Effects 0.000 description 5
- 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 4
- 241000218993 Begonia Species 0.000 description 4
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 4
- 206010061218 Inflammation Diseases 0.000 description 4
- 102100039019 Nuclear receptor subfamily 0 group B member 1 Human genes 0.000 description 4
- 229930003268 Vitamin C Natural products 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 235000006708 antioxidants Nutrition 0.000 description 4
- 210000004969 inflammatory cell Anatomy 0.000 description 4
- 230000004054 inflammatory process Effects 0.000 description 4
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 4
- 230000003647 oxidation Effects 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 235000019154 vitamin C Nutrition 0.000 description 4
- 239000011718 vitamin C Substances 0.000 description 4
- 239000012224 working solution Substances 0.000 description 4
- 102000004127 Cytokines Human genes 0.000 description 3
- 108090000695 Cytokines Proteins 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 3
- 102000003777 Interleukin-1 beta Human genes 0.000 description 3
- 108090000193 Interleukin-1 beta Proteins 0.000 description 3
- 230000003115 biocidal effect Effects 0.000 description 3
- 239000006227 byproduct Substances 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 206010009887 colitis Diseases 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 230000008595 infiltration Effects 0.000 description 3
- 238000001764 infiltration Methods 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 150000003254 radicals Chemical class 0.000 description 3
- NCEXYHBECQHGNR-UHFFFAOYSA-N sulfasalazine Natural products C1=C(O)C(C(=O)O)=CC(N=NC=2C=CC(=CC=2)S(=O)(=O)NC=2N=CC=CC=2)=C1 NCEXYHBECQHGNR-UHFFFAOYSA-N 0.000 description 3
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-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
- -1 DPPH free radical Chemical class 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 206010030113 Oedema Diseases 0.000 description 2
- 230000002292 Radical scavenging effect Effects 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 2
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 2
- 230000003044 adaptive effect Effects 0.000 description 2
- 230000003064 anti-oxidating effect Effects 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 239000003480 eluent Substances 0.000 description 2
- 230000002708 enhancing effect Effects 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 235000019253 formic acid Nutrition 0.000 description 2
- 210000002175 goblet cell Anatomy 0.000 description 2
- 230000002757 inflammatory effect Effects 0.000 description 2
- 230000008946 inflammatory intestinal reaction Effects 0.000 description 2
- 230000000968 intestinal effect Effects 0.000 description 2
- 244000144972 livestock Species 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000013641 positive control Substances 0.000 description 2
- 244000144977 poultry Species 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- LOGFVTREOLYCPF-KXNHARMFSA-N (2s,3r)-2-[[(2r)-1-[(2s)-2,6-diaminohexanoyl]pyrrolidine-2-carbonyl]amino]-3-hydroxybutanoic acid Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H]1CCCN1C(=O)[C@@H](N)CCCCN LOGFVTREOLYCPF-KXNHARMFSA-N 0.000 description 1
- OCZVHBZNPVABKX-UHFFFAOYSA-N 1,1-diphenyl-2-(2,4,6-trinitrophenyl)hydrazine;ethanol Chemical compound CCO.[O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1NN(C=1C=CC=CC=1)C1=CC=CC=C1 OCZVHBZNPVABKX-UHFFFAOYSA-N 0.000 description 1
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- 241000283690 Bos taurus Species 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 238000011740 C57BL/6 mouse Methods 0.000 description 1
- 241001092040 Crataegus Species 0.000 description 1
- 235000014493 Crataegus Nutrition 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- 108010074328 Interferon-gamma Proteins 0.000 description 1
- 102000008070 Interferon-gamma Human genes 0.000 description 1
- 108010002352 Interleukin-1 Proteins 0.000 description 1
- 102000000589 Interleukin-1 Human genes 0.000 description 1
- UETNIIAIRMUTSM-UHFFFAOYSA-N Jacareubin Natural products CC1(C)OC2=CC3Oc4c(O)c(O)ccc4C(=O)C3C(=C2C=C1)O UETNIIAIRMUTSM-UHFFFAOYSA-N 0.000 description 1
- 241001481296 Malus spectabilis Species 0.000 description 1
- 101001044384 Mus musculus Interferon gamma Proteins 0.000 description 1
- IOVCWXUNBOPUCH-UHFFFAOYSA-M Nitrite anion Chemical compound [O-]N=O IOVCWXUNBOPUCH-UHFFFAOYSA-M 0.000 description 1
- 206010034133 Pathogen resistance Diseases 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 102000007079 Peptide Fragments Human genes 0.000 description 1
- 108010033276 Peptide Fragments Proteins 0.000 description 1
- 101710188689 Small, acid-soluble spore protein 1 Proteins 0.000 description 1
- 101710188693 Small, acid-soluble spore protein 2 Proteins 0.000 description 1
- 101710166422 Small, acid-soluble spore protein A Proteins 0.000 description 1
- 101710166404 Small, acid-soluble spore protein C Proteins 0.000 description 1
- 101710174019 Small, acid-soluble spore protein C1 Proteins 0.000 description 1
- 101710174017 Small, acid-soluble spore protein C2 Proteins 0.000 description 1
- 101710174574 Small, acid-soluble spore protein gamma-type Proteins 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 102100036407 Thioredoxin Human genes 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 150000001450 anions Chemical class 0.000 description 1
- 238000003705 background correction Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000000112 colonic effect Effects 0.000 description 1
- 239000012228 culture supernatant Substances 0.000 description 1
- 230000006735 deficit Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- MGJZITXUQXWAKY-UHFFFAOYSA-N diphenyl-(2,4,6-trinitrophenyl)iminoazanium Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1N=[N+](C=1C=CC=CC=1)C1=CC=CC=C1 MGJZITXUQXWAKY-UHFFFAOYSA-N 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000002224 dissection Methods 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003640 drug residue Substances 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 238000013401 experimental design Methods 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 230000007760 free radical scavenging Effects 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 238000012826 global research Methods 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 239000000413 hydrolysate Substances 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 229960003130 interferon gamma Drugs 0.000 description 1
- 230000007358 intestinal barrier function Effects 0.000 description 1
- 210000004347 intestinal mucosa Anatomy 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 238000001819 mass spectrum Methods 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 210000004877 mucosa Anatomy 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 230000004792 oxidative damage Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000004007 reversed phase HPLC Methods 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 235000010288 sodium nitrite Nutrition 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- NCEXYHBECQHGNR-QZQOTICOSA-N sulfasalazine Chemical compound C1=C(O)C(C(=O)O)=CC(\N=N\C=2C=CC(=CC=2)S(=O)(=O)NC=2N=CC=CC=2)=C1 NCEXYHBECQHGNR-QZQOTICOSA-N 0.000 description 1
- 229960001940 sulfasalazine Drugs 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 208000037816 tissue injury Diseases 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 229940126680 traditional chinese medicines Drugs 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/08—Peptides having 5 to 11 amino acids
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
- A23K10/37—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms from waste material
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/105—Aliphatic or alicyclic compounds
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K40/00—Shaping or working-up of animal feeding-stuffs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Polymers & Plastics (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Food Science & Technology (AREA)
- Animal Husbandry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Biotechnology (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Botany (AREA)
- Physiology (AREA)
- Mycology (AREA)
- Molecular Biology (AREA)
- Gastroenterology & Hepatology (AREA)
- Immunology (AREA)
- Epidemiology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
本发明提供一种具有抗炎抗氧化功能的动物饲料添加剂，即一种中药类型的、具有抗生物功效的饲料添加剂，其制备方法是将干燥的油橄榄果渣粉样品和亚临界萃取剂混合，进行亚临界萃取，得到萃取体系；脱除所述萃取体系的亚临界萃取剂，得到脱除果渣油后的果渣粉体系；将所述果渣粉体系与乙醇混合，进行浸提，所得提取液经减压蒸馏，得到提取物；将所述提取物体系与石油醚混合，静置分层后取石油醚相，所得石油醚相过滤制成的。本发明所提取的山楂酸和齐墩果酸，具有抗炎和抗氧化的功效，经动物试验证实，其可以短时间增强肠道健康，添加了多肽之后，改善了山楂酸和齐墩果酸长其使用引起的副作用，可作为新型动物饲料添加剂推广使用。
The present invention provides an animal feed additive with anti-inflammatory and antioxidant functions, that is, a Chinese medicine type feed additive with anti-biological effects. The preparation method is as follows: a dried olive pomace powder sample is mixed with a subcritical extractant, and subcritical extraction is performed to obtain an extraction system; the subcritical extractant of the extraction system is removed to obtain a pomace powder system after pomace oil is removed; the pomace powder system is mixed with ethanol, and leaching is performed, and the obtained extract is subjected to reduced pressure distillation to obtain an extract; the extract system is mixed with petroleum ether, and the petroleum ether phase is taken after standing and stratifying, and the obtained petroleum ether phase is filtered to prepare. The maslinic acid and oleanolic acid extracted by the present invention have anti-inflammatory and antioxidant effects, and animal experiments have confirmed that they can enhance intestinal health in a short time. After adding polypeptides, the side effects caused by the long use of maslinic acid and oleanolic acid are improved, and the present invention can be promoted and used as a new animal feed additive.
Description
Technical Field
The invention relates to the technical field of biological products, in particular to an animal feed additive with anti-inflammatory and antioxidant functions.
Background
Antibiotics are substances obtained via microbial culture or chemical synthesis and have the ability to specifically kill microorganisms. In 1928, the uk scientist alexandrite flemings discovered penicillin for the first time, which marks the entrance of man into the antibiotic era. Subsequently, in 1946, the us scientist Moore et al first discovered that antibiotics had a growth promoting effect in broiler feed, and this discovery led the global livestock industry to enter a new era of antibiotic growth promotion. Since then, antibiotics have been widely used as feed additives for feeding various animals such as pigs, poultry, cattle, sheep, aquatic animals, and the like. The antibiotics with the dosage lower than the therapeutic dosage are added into the feed, so that the growth of animals can be promoted, the feed conversion rate can be improved, the morbidity and mortality of livestock and poultry can be reduced, and the important promotion effect on the development of the breeding industry can be realized. However, the long-term addition of antibiotics also brings a series of problems such as bacterial resistance, environmental pollution, drug residues in animal-derived foods, public health potential safety hazards and the like.
The prohibition of the use of feeding antibiotics in animal production has become a consensus and an irreversible trend worldwide. The use of antibiotics as feed additives has been totally banned by the european union in 2006. And the agricultural rural department of China also announced No. 194 in 9 of 2019, announced that all the drug feed additive varieties with the growth promoting function except the traditional Chinese medicines are stopped from 1 of 2020, which marks the new era of no feed resistance of the animal husbandry industry of China.
With the deep advancement of the forbidden process of the global feeding antibiotics, the search for safe and effective alternatives has become a hotspot of global research, and the market has urgent demands for safe and efficient feeding antibiotic alternative products. The feed plant and the additive thereof are considered as one of ideal substitutes for feed antibiotics in animal production because of the advantages of rich resources, natural sources, comprehensive functions, high safety, low toxic and side effects, difficult generation of drug resistance, no (or low) residues and the like.
Disclosure of Invention
The invention aims to provide an animal feed additive with anti-inflammatory and antioxidant functions, namely a traditional Chinese medicine type feed additive with an anti-biological effect.
The preparation method of the animal feed additive provided by the invention comprises the following steps:
mixing a dried olive fruit residue powder sample with a subcritical extractant, and performing subcritical extraction to obtain an extraction system;
Removing subcritical extractant of the extraction system to obtain a residue powder system after removing the fruit residue oil;
Mixing the fruit residue powder system with ethanol, leaching, and distilling the obtained extract under reduced pressure to obtain extract;
Mixing the extract system with petroleum ether, stirring, standing for layering, collecting petroleum ether phase, and filtering the obtained petroleum ether phase to obtain filtrate.
The method comprises the steps of drying and crushing olive pomace, wherein the water content of the dried material is less than or equal to 5%, and the particle size of the crushed material is 20-40 meshes.
Preferably, the subcritical extractant comprises one or more of butane, propane, isobutane or dimethyl ether.
Preferably, the subcritical extraction is carried out at a pressure of 0.45-0.8 MPa, a temperature of 35-45 ℃ and a time of 40-45 min.
Preferably, the subcritical extractant of the extraction system is removed by reduced pressure distillation, wherein the temperature of the reduced pressure distillation is 45-50 ℃, the vacuum degree is less than or equal to 0.5MPa, and the time is 25-30 min.
Preferably, the volume ratio of the extract system to the petroleum ether is 1:1-1:20.
Preferably, the purification is performed under the condition of centrifugation, wherein the rotation speed of the centrifugation is 6000-20000 r/min, and the time is 5-20 min.
Furthermore, the animal feed additive is also added with polypeptide for preventing and treating enteritis;
the polypeptide is extracted from begonia;
furthermore, the polypeptide has a sequence DKQGEP (SEQ ID NO: 1).
Animal experiments prove that the crataegolic acid and oleanolic acid extracted by the invention have the effects of resisting inflammation and oxidization, can strengthen intestinal health in a short time, improve side effects caused by the growth of crataegolic acid and oleanolic acid after adding polypeptide, and can be popularized and used as novel animal feed additives.
Drawings
FIG. 1 shows the clearance of DPPH free radical by crataegolic acid and oleanolic acid;
FIG. 2 shows the free radical scavenging ability of crataegolic acid and oleanolic acid against ABTS +;
FIG. 3 is the effect of crataegolic acid and oleanolic acid on NO release;
FIG. 4 is a reversed phase high performance liquid chromatography;
FIG. 5 is a secondary mass spectrum of a selected polypeptide;
FIG. 6H & E pathology observations of the mouse colon;
FIG. 7 IL-4, IL-1. Beta., TNF-. Alpha. And IFN-. Gamma.levels in serum of mice of each group;
FIG. 8 is a graph showing the effect of crataegolic acid and oleanolic acid on the oxidation index of mice with enteritis.
Detailed Description
It has been found that natural organic substances such as crataegolic acid, oleanolic acid and some pentacyclic triterpenic acids contained in food processing byproducts such as olive pomace which can be used as feeds have good anti-inflammatory and antioxidant effects and intestinal health enhancing effects. Therefore, the olive pomace is used as a raw material to research and prepare processing byproducts containing functional substances such as anti-inflammatory, antioxidant and intestinal health enhancing substances, and the processing byproducts can be used as animal feed additives.
The present invention will be described in detail with reference to specific embodiments and drawings.
Example 1 extraction of crataegolic acid and oleanolic acid
Drying olive pomace until the water content is not higher than 5%, crushing by a crusher, sieving by a 40-mesh sieve and a 20-mesh sieve successively, crushing the olive pomace with the particle size of 20-40 meshes, extracting by a subcritical extractant at the temperature of 35-45 ℃ under the pressure of 0.45-0.8 MPa for 40-45 min, removing the subcritical extractant to obtain pomace powder, mixing the pomace powder with ethanol, leaching, distilling the extract under reduced pressure to obtain an extract, mixing and stirring a system of the obtained extract with petroleum ether according to the volume ratio of 1:1-1:20, standing and layering, obtaining a petroleum ether phase, filtering the petroleum ether phase, and centrifuging the obtained filtrate at the rotating speed of 6000-20000 r/min to obtain hawthornic acid and oleanolic acid.
Example 2 determination of anti-inflammatory and antioxidant effects of Crataegus acid and Oleanolic acid
1. Determination of the clearance of DPPH free radicals
Measuring 0.1mL of crataegolic acid and oleanolic acid sample solution, adding the solution into a 10mL volumetric flask, diluting the solution to be close to 1mL of scale marks by distilled water, adding 3.5mL of DPPH ethanol solution with the concentration of 0.2mmol/L, and continuously metering the volume to the scale marks of the volumetric flask by ethanol to ensure uniform mixing of the solutions. The mixture was placed in a dark environment for reaction for 30min. Thereafter, the absorbance value of the solution was measured at a wavelength of 517 nm. Meanwhile, a DPPH solution without a sample and a sample solution without DPPH were prepared separately, and the absorbance values of each were measured at the same wavelength to be used as a blank control and a sample background correction. In addition, the same concentration of Vitamin C (VC) solution was used as a positive control, and the absorbance was measured by the same treatment. Finally, the DPPH radical scavenging rate of the sample is calculated according to the obtained data.
Wherein A 1 is the absorbance value of the sample solution after reaction, A 2 is the absorbance value of the solution without DPPH, and A 0 is the absorbance value of the solution without sample.
As can be seen from fig. 1, the scavenging effect of the crataegolic acid and oleanolic acid solution on DPPH free radicals is close to VC, indicating that the crataegolic acid and oleanolic acid solution have good oxidation resistance.
2. Determination of the radical scavenging Capacity of ABTS +
Respectively measuring 0.5mL of sample solution into two 10mL volumetric flasks, adding 2.5mL of ABTS + working solution, and accurately diluting to a scale mark by distilled water to ensure uniform mixing. After the reaction was allowed to stand at room temperature for 10 minutes without light, the absorbance of each solution was measured at a wavelength of 734nm using a spectrophotometer. Meanwhile, two control groups were set, one group containing only ABTS + working solution (no sample added) to determine its background absorbance, and the other group containing only sample solution (no ABTS + working solution added) to determine its own absorbance, all measured at the same wavelength. Finally, the VC solution with the same concentration level is used as a positive control, and the free radical clearance of the sample is calculated according to the obtained data.
Wherein A 1 is the absorbance value of the sample solution after reaction, A 2 is the absorbance value of the solution without the addition of ABTS + working solution, and A 0 is the absorbance value of the solution without the addition of the sample.
As shown in FIG. 2, the crataegolic acid and oleanolic acid have good removal effect on ABTS +, and the removal rate is more than 70%.
3. NO release amount detection
The experimental design contained three groups, a blank control group (containing only DMEM medium, without cells), an LPS model group (LPS concentration 1.0. Mu.g/mL), and an experimental group (LPS concentration 1.0. Mu.g/mL combined with 50. Mu.g/mL each of crataegolic acid and oleanolic acid). Each set of samples was added to the culture wells in a volume of 100. Mu.L and then incubated for 24 hours in a CO 2-containing incubator at 37 ℃. The nitrite (NaNO 2) standard was diluted with the medium, 50. Mu.L of the cell culture supernatant was collected from each well, and after leaving at room temperature for 5 minutes, equal amounts of Griess reagent I (50. Mu.L) and Griess reagent II (50. Mu.L) were sequentially added. The amount of NO produced in each group was precisely calculated by measuring the optical density value (OD) at 540nm wavelength in combination with the NaNO2 standard curve.
NO acts as a key regulator and plays an important role in various physiological functions and inflammatory processes, and particularly has a remarkable effect in an acute inflammatory model of RAW264.7 macrophages induced by LPS. FIG. 3 shows the effect of maslinic acid and oleanolic acid on NO secretion in the model, wherein the NO level in the model group is remarkably increased (P < 0.01) compared with the control group after LPS (1.0 mug/mL) is treated for 24 hours, so that the effectiveness of an inflammation model is verified, and the NO content is remarkably reduced (P < 0.05) compared with the model group after maslinic acid and oleanolic acid are added, so that the maslinic acid and oleanolic acid have good anti-inflammatory effect.
The results of applying the crataegolic acid and the oleanolic acid as feed additives to the feeding of mice show that after the continuous feeding of the feed additives added with the crataegolic acid and the oleanolic acid for 2 months, the mice in the feeding group are found to have enteritis phenomenon, and the continuous feeding of the feed additives of the crataegolic acid and the oleanolic acid is suspected to cause enteritis diseases of fed animals, so that the feed prepared by taking the crataegolic acid and the oleanolic acid as the feed additives is more suitable for short-term treatment.
Example 3 extraction of polypeptide having enteritis preventing and treating action
Grinding Begonia tissue into homogenate, adding ddH 2 O, mixing, adding 3000U/mg trypsin, adjusting pH to 7.5, maintaining the temperature at 50deg.C, hydrolyzing for 5 hr, inactivating enzyme in a water bath at 100deg.C for 15min, centrifuging at 10000r/min for 10min, separating and collecting supernatant, concentrating, and drying to obtain Begonia polypeptide dry powder.
The method comprises the steps of performing primary purification on the begonia polypeptide after enzymolysis by using an anion exchanger DEAE-52, selecting 20mM sodium acetate as a buffer solution, using Tris-HCl buffer solution containing 0-2mol/L NaCl as an eluent, then performing purification by using an HPLC method, purifying hydrolysate by using a reversed phase column 300SB-C18 (4.6X250 mM,5 μm, agilent), respectively using Acetonitrile (ACN) containing 0.1% Formic Acid (FA) and ACN as eluent x and y, setting the flow rate to be 0.5mL/min, performing elution by using the following sequence, namely 1-5min,100% x, 5-40min,5-90% y (linear gradient), 40-50min,100% x, detector wavelength of 280nm, and performing activity prediction by using a mass spectrometer, and inputting the obtained polypeptide sequence into a BIOPEP-Ubioactive peptide database to screen out the peptide fragments with high activity, wherein the sequence is DHTLP (polypeptide a), IDFLEH (polypeptide b) and DKQGEP (polypeptide C).
The three selected polypeptide sequences with the best activity are sent to a biological engineering (Shanghai) Co., ltd for synthesis, RAW264.7 cells are cultivated in a complete medium containing 10% fetal calf serum at 37 ℃ and 5% CO 2, and after every 24 hours passage, the cells in logarithmic growth phase are taken, different polypeptides are added for cultivation for 2 days, 20 mu L of MTT solution is added for continuous cultivation for 3 hours, and the anti-inflammatory performance in vitro is measured by ELISA method.
As can be seen from the results in Table 1, polypeptide c has the best effect on TNF- α and IL-1β clearance, indicating that it has the best anti-inflammatory properties in vitro and its sequence is DKQGEP (SEQ ID NO: 1).
TABLE 1 in vitro anti-inflammatory Activity of three Malus spectabilis polypeptides selected (clearance/%)
40 Male C57BL/6 mice of 6 weeks of age were used, all weighing in the range of (20.+ -.3 g). One week of adaptive feeding was performed during which all groups were given normal drinking water and feed. Mice were divided into four groups of 10 mice each, normal, model, control and test. After the end of the adaptive feeding, the other three groups of mice were continuously and freely drunk 2% crataegolic acid and oleanolic acid solution, except for the normal group. Meanwhile, mice in the control group were also perfused with 100mg/kg of sulfasalazine daily, while mice in the test group were perfused with the polypeptide-added crataegolic acid and oleanolic acid solution daily. After 60 days, mice were fasted, but not water, for 12 hours. Thereafter, the mice were dissected, blood samples were collected through orbital veins, and during the dissection process, blood samples and colon tissue were collected for H & E staining and pathological observations were made.
Under the induction of hawthornic acid and oleanolic acid, colon tissues show obvious lesion characteristics including destruction of intestinal epithelium, loss of goblet cells, strong inflammatory cell infiltration, and disappearance of crypts. As shown in fig. 6, the colon structure of normal mice remained intact, crypts were clearly visible, goblet cells were enriched, and no evidence of significant injury was observed. In contrast, the colonic mucosa of the model group mice was severely damaged, the crypt epithelial cell morphology was distorted, and infiltration of a large number of inflammatory cells was accompanied, showing a severe impairment of intestinal barrier function. While the control group showed relatively complete colon structure, it was observed that some inflammatory cells infiltrate and intestinal wall edema occurred. Notably, in colon tissue sections of the mice of the test group, crypt structures were clearly visible with little edema or inflammatory cell infiltration. The results prove that the crataegolic acid and the oleanolic acid added with the polypeptide c can effectively reduce the tissue injury of the crataegolic acid and the oleanolic acid to the colon of the mouse, and the protection effect is more obvious compared with SASP.
Example 4 alleviation of mice on enteritis by the combination of polypeptide and crataegolic acid
Mice were screened using the criteria of example 3, randomly divided into 4 groups, normal and test groups (model, test one, test two), wherein the test groups were filled with 2% DSS water daily, the test one group was also filled with 100mg/kg crataegolic acid and oleanolic acid daily, the test two groups were mixed with crataegolic acid oleanolic acid, and the normal groups were left untreated, and after 28 days mice were sacrificed by cervical removal, the intestinal tracts of the mice were taken, and colon tissues were analyzed for expression of related genes and cytokines.
1. Determination of inflammatory factor index of mice
The content of interleukin-4 (IL-4) tumor necrosis factor-alpha (TNF-alpha), interleukin-1 beta (IL-1 beta) and mouse interferon gamma (IFN-gamma) in the serum of the mouse is detected by using the kit.
Cytokine levels in mice are shown in figure 7. TNF- α, IL-1 β, IFN- γ levels were significantly elevated (p < 0.05) in both the test and test groups compared to the model group, and the data in the test group was closer to the normal group. IL-4 is an important anti-inflammatory cytokine, and it can be seen from FIG. 7 that IL-4 levels were significantly reduced (p < 0.05) in the model group compared to the normal group, while IL-4 levels were significantly increased in the test one and test two groups. The research proves that the crataegolic acid and the oleanolic acid can reduce the intestinal inflammation induced by DSS, the anti-inflammatory effect of the crataegolic acid and the oleanolic acid solution added with the polypeptide c is better than that of the crataegolic acid and the oleanolic acid which are singly used, the intestinal inflammatory reaction is obviously reduced after the crataegolic acid and the oleanolic acid are used, and the anti-inflammatory effect is obviously enhanced, thereby achieving the purpose of reducing the intestinal inflammatory reaction.
2. Determination of Oxidation index in mouse serum
The method comprises the steps of measuring Myeloperoxidase (MPO) in mouse serum by an enzyme-linked immunosorbent assay (ELISA), measuring the content of Catalase (CAT), glutathione Reductase (GR) and Malondialdehyde (MDA) in the mouse serum by a micro-method, and measuring the content of superoxide dismutase (SOD) in the mouse serum by an enzyme-labeled method.
In DSS-induced colitis in mice, oxidative stress in the colon tissue is an important indicator, and studies indicate that colitis is often accompanied by an increase in oxidative stress, so improving oxidative stress in colon tissue is a potential measure for reducing the occurrence of colitis. As shown in figure 8, which shows SOD, CAT, GR, MDA, MPO levels in mouse serum, CAT (P < 0.05), SOD (P < 0.05), GR (P < 0.05) levels were significantly reduced in model mice compared to normal mice, whereas MDA (P < 0.05). MPO (p < 0.05) levels were significantly elevated, thus demonstrating that DSS produced oxidative stress in mice, causing oxidative damage to colon tissue in mice. After the intervention of the crataegolic acid and the oleanolic acid is added, oxidation indexes show opposite trend with a model group, CAT and GR levels in mice are obviously improved (p < 0.05), SOD levels are also improved, MDA (p < 0.05) and MPO (p < 0.05) levels are reduced, the intervention effect of the crataegolic acid and the oleanolic acid added with the polypeptide c is better, the colon oxidative stress of the mice caused by the crataegolic acid and the oleanolic acid is indicated, and the antioxidation effect is stronger after the polypeptide c is added.
The crataegolic acid and oleanolic acid extracted by the invention have the effects of anti-inflammation and anti-oxidation, are used as short-term treatment, improve enteritis caused by long-term use after the polypeptide c is added, strengthen intestinal health, and can be applied and popularized as a natural feed additive.
Claims (10)
1. An animal feed additive, characterized in that the animal feed additive ethical preparation method comprises the following steps:
mixing a dried olive fruit residue powder sample with a subcritical extractant, and performing subcritical extraction to obtain an extraction system;
Removing subcritical extractant of the extraction system to obtain a residue powder system after removing the fruit residue oil;
Mixing the fruit residue powder system with ethanol, leaching, and distilling the obtained extract under reduced pressure to obtain extract;
Mixing the extract system with petroleum ether, stirring, standing for layering, collecting petroleum ether phase, and filtering the obtained petroleum ether phase to obtain filtrate as animal feed additive.
2. The animal feed additive according to claim 1, wherein the olive pomace powder sample is subjected to pretreatment before being mixed with the subcritical extractant, and the pretreatment comprises the steps of drying and crushing olive pomace, wherein the water content of the dried material is less than or equal to 5%, and the particle size of the crushed material is 20-40 meshes.
3. An animal feed additive as claimed in claim 1 or claim 2 wherein the subcritical extractant comprises one or more of butane, propane, isobutane or dimethyl ether.
4. The animal feed additive of claim 1, wherein the subcritical extraction is carried out at a pressure of 0.45-0.8 mpa, a temperature of 35-45 ℃ and a time of 40-45 min.
5. The animal feed additive according to claim 1, wherein the subcritical extractant removing mode of the extraction system is reduced pressure distillation, the temperature of the reduced pressure distillation is 45-50 ℃, the vacuum degree is less than or equal to 0.5MPa, and the time is 25-30 min.
6. The animal feed additive of claim 1, wherein the extract system and petroleum ether are present in a volume ratio of 1:1 to 1:20.
7. The animal feed additive of claim 1, wherein the animal feed additive further comprises a polypeptide for preventing and treating enteritis.
8. The animal feed additive of claim 7, wherein the polypeptide has an amino acid sequence of SEQ ID No. 1.
9. An animal feed, wherein the animal feed is supplemented with the animal feed additive of claim 1.
10. The animal feed of claim 9, wherein the animal feed further comprises a polypeptide having the amino acid sequence of SEQ ID No. 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202411224968.2A CN119097046A (en) | 2024-09-03 | 2024-09-03 | An animal feed additive with anti-inflammatory and antioxidant functions |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202411224968.2A CN119097046A (en) | 2024-09-03 | 2024-09-03 | An animal feed additive with anti-inflammatory and antioxidant functions |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN119097046A true CN119097046A (en) | 2024-12-10 |
Family
ID=93709913
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202411224968.2A Pending CN119097046A (en) | 2024-09-03 | 2024-09-03 | An animal feed additive with anti-inflammatory and antioxidant functions |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN119097046A (en) |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040185157A1 (en) * | 2001-09-28 | 2004-09-23 | The Nisshin Oillio, Ltd. | Method for preparing oil and fat compositions comprising oleanolic acid and/or maslinic acid |
| CN101805664A (en) * | 2010-03-29 | 2010-08-18 | 甘肃省轻工研究院 | Method and device for extracting olive pomace oil from olive pomace |
| CN103169771A (en) * | 2011-12-23 | 2013-06-26 | 中国科学院兰州化学物理研究所 | Method for extracting maslinic acid and oleanolic acid containing mixture from olea europaea L. pomace |
| CN107602655A (en) * | 2017-09-27 | 2018-01-19 | 北京林业大学 | The Subcritical water chromotagraphy and separation method of oleanolic acid in a kind of response phase method optimization olive pomace |
| CN107969552A (en) * | 2017-12-24 | 2018-05-01 | 中国林业科学研究院林产化学工业研究所 | A kind of extractive of olive growing leaves feed addictive and its application |
| CN110872337A (en) * | 2019-12-06 | 2020-03-10 | 湖南先伟实业有限公司 | Method for simultaneously preparing different high-purity triterpenic acids from rosemary |
| CN115361877A (en) * | 2020-03-31 | 2022-11-18 | 卢斯塔公司 | Feed additives |
| CN117815294A (en) * | 2023-11-02 | 2024-04-05 | 中国科学院兰州化学物理研究所 | Olive pomace extract rich in anti-inflammatory active ingredient |
| US20240139291A1 (en) * | 2021-11-25 | 2024-05-02 | Industry-Academic Cooperation Foundation, Yonsei University | Collagen hydrolysate comprising gpcr ligand peptide |
-
2024
- 2024-09-03 CN CN202411224968.2A patent/CN119097046A/en active Pending
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040185157A1 (en) * | 2001-09-28 | 2004-09-23 | The Nisshin Oillio, Ltd. | Method for preparing oil and fat compositions comprising oleanolic acid and/or maslinic acid |
| CN101805664A (en) * | 2010-03-29 | 2010-08-18 | 甘肃省轻工研究院 | Method and device for extracting olive pomace oil from olive pomace |
| CN103169771A (en) * | 2011-12-23 | 2013-06-26 | 中国科学院兰州化学物理研究所 | Method for extracting maslinic acid and oleanolic acid containing mixture from olea europaea L. pomace |
| CN107602655A (en) * | 2017-09-27 | 2018-01-19 | 北京林业大学 | The Subcritical water chromotagraphy and separation method of oleanolic acid in a kind of response phase method optimization olive pomace |
| CN107969552A (en) * | 2017-12-24 | 2018-05-01 | 中国林业科学研究院林产化学工业研究所 | A kind of extractive of olive growing leaves feed addictive and its application |
| CN110872337A (en) * | 2019-12-06 | 2020-03-10 | 湖南先伟实业有限公司 | Method for simultaneously preparing different high-purity triterpenic acids from rosemary |
| CN115361877A (en) * | 2020-03-31 | 2022-11-18 | 卢斯塔公司 | Feed additives |
| US20240139291A1 (en) * | 2021-11-25 | 2024-05-02 | Industry-Academic Cooperation Foundation, Yonsei University | Collagen hydrolysate comprising gpcr ligand peptide |
| CN117815294A (en) * | 2023-11-02 | 2024-04-05 | 中国科学院兰州化学物理研究所 | Olive pomace extract rich in anti-inflammatory active ingredient |
Non-Patent Citations (1)
| Title |
|---|
| 王着: "油橄榄果渣中山楂酸和齐墩果酸提取分离工艺研究", 中国优秀硕士论文电子期刊医药卫生科技, 15 November 2012 (2012-11-15), pages 1 - 70 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Lau et al. | Ethnomedicinal uses, pharmacological activities, and cultivation of Lignosus spp.(tiger׳ s milk mushrooms) in Malaysia–A review | |
| Xiang et al. | Protective effects of shrimp peptide on dextran sulfate sodium-induced colitis in mice | |
| CN108030089B (en) | Method for recycling salvia miltiorrhiza water extraction and alcohol precipitation herb residues | |
| Sipping et al. | Antioxidant and anti-inflammatory activities of Ganoderma resinaceum (Boud) fruiting bodies extracts | |
| Han et al. | Hericium erinaceus polysaccharide improves the microstructure, immune function, proliferation and reduces apoptosis of thymus and spleen tissue cells of immunosuppressed mice | |
| CN1446785A (en) | Method of producing magnolol and honokiol | |
| CN103564495B (en) | Propolis and cordycepic hypha powder composition, as well as preparation and application thereof | |
| CN116004454A (en) | Application of Lactobacillus plantarum J26 and its composition in alleviating acute-on-chronic alcoholic liver injury | |
| TW201515654A (en) | Extraction process for caulerpa lentillifera extract and pharmaceutical composition containing caulerpa lentillifera extract for inhibiting growth of prostate cancer cells | |
| CN119097046A (en) | An animal feed additive with anti-inflammatory and antioxidant functions | |
| CN111714519B (en) | Application and composition of organic solvent extract of sea cockroach | |
| CN114656522B (en) | Antioxidant peptide and preparation method and application thereof | |
| CN106692206A (en) | Application of Xylaria nigripes mycelium extract in protecting nerve cells and composition for protecting nerve cells | |
| CN102895321A (en) | Preparation method of plantain herb polyphenol with antioxidant activity | |
| CN111793105A (en) | Extraction and application of protein from Alkali larvae by Alkali Fusion | |
| CN106916206A (en) | Hard clam polypeptide and preparation method and application | |
| CN1261452C (en) | Sugar latticeing material CHB extracted from serum of turtle or/and tortoise, preparation process and application in pharmacy thereof | |
| AU2021104232A4 (en) | Application of sanghuangporus sanghuang fermentation broth polysaccharides in medicaments of anti-avian influenza virus | |
| CN110257167A (en) | Poplar bark lipoid supercritical extraction method and application | |
| CN117586889B (en) | Ma Lem-calix palmatum and dinoflagellate mutant strain with high polysaccharide yield and application thereof | |
| CN113893214B (en) | Application of rhynchophylla extract in cosmetics | |
| CN114984064B (en) | Morchella fruiting body active site and its preparation method and application | |
| CN113598280B (en) | Mouse feed containing selenium-enriched magnolia bark extract and application of mouse feed in construction of fatty liver symptom-free obese mouse model | |
| CN110551673B (en) | Method for improving fucoxanthin content by treating phaeodactylum tricornutum with rapamycin | |
| CN118452307A (en) | Preparation method of solid fermentation product of Astragalus and application thereof in preparation of feed additive for regulating growth and development of weaned donkey foals |
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 |