CN109966373A - A kind of application of black fruit fructus lycii seed oil extract in blood lipid-lowering medicine - Google Patents
A kind of application of black fruit fructus lycii seed oil extract in blood lipid-lowering medicine Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/81—Solanaceae (Potato family), e.g. tobacco, nightshade, tomato, belladonna, capsicum or jimsonweed
- A61K36/815—Lycium (desert-thorn)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/06—Antihyperlipidemics
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- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
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- Microbiology (AREA)
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- Alternative & Traditional Medicine (AREA)
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- Diabetes (AREA)
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Abstract
The present invention relates to field of biotechnology, and in particular to a kind of application of black fruit fructus lycii seed oil extract in blood lipid-lowering medicine.The black fruit fructus lycii seed oil extract fatty acid and sterol can be used as effective component and reducing blood lipid pharmaceutical formulation are made with pharmaceutically acceptable any carrier according to a conventional method.Black fruit fructus lycii seed oil extract fatty acid of the invention and sterol can significantly inhibit with hyperlipidemia related target pancreatic lipase and cholesterol esterase activity, and zebra fish and mouse experiment also demonstrate that black fruit fructus lycii seed oil fatty acid and sterol blood fat reducing function are obvious；Black fruit fructus lycii seed oil fatty acid and sterol have the function of good reducing blood lipid as hypolipidemic activity ingredient, and its obvious effect, the mechanism of action understand, target spot is clear, meet the requirement of pharmaceutical preparation exploitation.
Description
Technical field
The present invention relates to field of biotechnology, and in particular to a kind of black fruit fructus lycii seed oil extract is in blood lipid-lowering medicine
Using.
Background technique
Hyperlipidemia refers in blood TC or TG and its associated lipoprotein level is excessively high or serum high-density LP gallbladder
Sterol is too low.Hyperlipidemia is related to a variety of diseases such as hypertension, coronary heart disease, cerebral apoplexy, diabetes.As caused by hyperlipidemia
Atherosclerosis is the main reason for causing the cardiovascular and cerebrovascular diseases such as coronary heart disease, myocardial infarction, cerebral hemorrhage, cerebral infarction, and mesh
Preceding fat-reducing medicament on the market mainly has Statins, fibrates, niacin class etc..Statins are two phthaleins of typical light methylpent
CoA reductase inhibitor, it is significant in efficacy but expensive, and there is the potential toxic side effects such as hepatotoxicity wind agitation, muscle poison.Cause
This, the drug for developing natural, nontoxic treatment hyperlipidemia will have important practical significance and potentiality to be exploited.
Black fruit fructus lycii (Lycium ruthenicum Murr.) is Solanaceae Lycium, is commonly called as lycium ruthenicum, is China west
Northern region-by-region common wild bush is circle after berry fruit is mature, darkly purple.Tibetan medicine black fruit fructus lycii is referred to as " other
Ma ", the Four-Volume Medical Code are recorded, and Tibetan medicine is used as medicine with its ripening fruits, the diseases such as treatment cardiopyretic disease, heart disease, irregular menstruation, menopause
Disease, " Uygur medicine will " are recorded, and lycium ruthenicum fruit and root skin can treat the diseases such as urethral calculus, tinea scabies, gingiva bleeding gingival hemorrhage, it is civil then
As strengthening by means of tonics, improving eyesight and depressor.It is rich in anthocyanin in black fruit fructus lycii ripe berry, belongs to typical natural anthocyanin
Class phytochrome resource.Compared with red fructus lycii, the nutrient composition contents such as vitamin, minerals are richer, the flower especially contained
Color glycosides, medicinal and health value is very high, is known as " soft gold ".
At present in terms of the research to lycium ruthenicum extract as medicinal application: Chinese patent ZL201510711969.4 is disclosed
A kind of black fruit fructus lycii health-care tablet production method for alleviating asthenopia, can be effective with the black fruit fructus lycii health-care tablet that this method produces
Alleviate eye strain；Granted patent ZL201310274771.5 disclose a kind of black Fructus Lycii extract and preparation method and
Using the black Fructus Lycii extract can be used for preparing the drug that the leucocyte for the treatment of human body reduces；Granted patent ZL
201210516648.5 disclose a kind of black fruit fructus lycii alkali-extracted polysaccharide and its preparation and immunosupress can be improved in application, the polysaccharide
The nospecific immunity of mouse, cellular immunity, humoral immune function, inhibition of effective antagonism cyclophosphamide to immune function of mice
Effect, can be used for preparing immunoregulation medicament and health care product；For another example granted patent ZL201410064343.4 invented it is a kind of with
The plants compatibility such as black fruit fructus lycii leaf and turmeric, lepidium seed for reducing blood lipid Chinese medicinal granule.Black fruit fructus lycii seed oil extract is made
For the application in blood lipid-lowering medicine, reported so far there is not yet disclosing.
Summary of the invention
Technical problem to be solved by the invention is to provide a kind of black fruit fructus lycii seed oil to prepare the application in blood lipid-lowering medicine.
Based on the above issues, the purpose of the present invention is to provide a kind of black fruit fructus lycii seed oil extracts in blood lipid-lowering medicine
Application.
The application of a kind of black fruit fructus lycii seed oil extract in blood lipid-lowering medicine, wherein the black fruit fructus lycii seed oil mentions
Object is taken to apply in blood lipid-lowering medicine as effective component.
Further, the black fruit fructus lycii seed oil extract can be used as effective component according to a conventional method with can pharmaceutically connect
Reducing blood lipid pharmaceutical formulation is made in any carrier received.
Further, the fruit Boxthorn Seed Oil extract is fatty acid and sterol.
Further, the fruit Boxthorn Seed Oil extract fat acid and sterol pass through supercritical fluid extraction.
Compared with the prior art, the invention has the following beneficial effects:
(1) black fruit fructus lycii seed oil of the present invention is a kind of pure natural, plant source, nontoxic non-stimulated substance, is extracted
Object fatty acid and sterol can significant reducing blood lipid active constituent.
(2) black fruit fructus lycii seed oil extract fatty acid and sterol of the present invention can significantly inhibit and hyperlipidemia related target pancreas rouge
Fat enzyme and cholesterol esterase activity, and zebra fish and mouse experiment also demonstrate, black fruit fructus lycii seed oil fatty acid and sterol pair
The zebra fish hyperlipidemia that yolk powder induces has effect for reducing blood fat, while can significantly reduce total cholesterol in mice serum high in fat
(TC), triglycerides (TG), low-density lipoprotein (LDL) is horizontal, and increasing high density lipoprotein (HDL) is horizontal.
(3) for pancreatic lipase in three ester hydrolysis of gastrointestinal tract catalyzing glycerol, triglycerides is the energy source of high calorie, is inhibited
The absorption of triglycerides can the diseases such as pre- preventing obesity and hyperlipidemia, inhibit pancreatic lipase activity be pre- preventing obesity main road
Diameter.Cholesterol esterase is widely studied as the target of control cholesterol levels.Cholesterol esterase is in hydrolysis diet cholesterol
It plays an important role in terms of ester and transhipment free cholesterol to enterocyte.Black fruit fructus lycii seed oil fatty acid and sterol are to pass through inhibition
The activity of cholesterol esterase and pancreatic lipase is to have the function that reducing blood lipid.Therefore, black fruit fructus lycii seed oil of the present invention mentions
It takes object fatty acid and sterol as hypolipidemic activity ingredient, has the function of good reducing blood lipid, and its obvious effect, effect
Mechanism understands, target spot is clear, meets the requirement of pharmaceutical preparation exploitation.
Detailed description of the invention
Lipid-lowering effect figure in Fig. 1 black fruit fructus lycii seed oil extract fatty acid of the present invention and sterol zebra fish body.
Fig. 2 black fruit fructus lycii seed oil extract fatty acid of the present invention and sterol reduce TC effect picture in mice serum.
Fig. 3 is that black fruit fructus lycii seed oil extract fatty acid of the present invention and sterol reduce TG effect picture in mice serum.
Fig. 4 is that black fruit fructus lycii seed oil extract fatty acid of the present invention and sterol increase HDL effect picture in mice serum.
Fig. 5 is that black fruit fructus lycii seed oil extract fatty acid of the present invention and sterol reduce LDL effect picture in mice serum.
Fig. 6 is that black fruit fructus lycii seed oil extract fatty acid of the present invention and sterol inhibit pancreatic lipase activity figure.
Fig. 7 is that black fruit fructus lycii seed oil extract fatty acid of the present invention and sterol inhibit cholesterol esterase activity figure.
Fig. 8 is that black fruit fructus lycii seed oil extract fatty acid of the present invention and sterol inhibit cholesterol esterase mechanism choice.
Fig. 9 is that black fruit fructus lycii seed oil extract fatty acid of the present invention and sterol inhibit cholesterol esterase power figure.
Specific embodiment
Embodiment 1
A kind of application of black fruit fructus lycii seed oil extract in blood lipid-lowering medicine.
The blood lipid-lowering medicine containing black fruit fructus lycii seed oil extract fatty acid and sterol effective component is prepared first:
The black fruit fructus lycii seed oil extract fatty acid and sterol with 80% additive amount be prepared into can reducing blood lipid soft capsule preparation.
Specific preparation method are as follows: by 80 parts of black fruit fructus lycii seed oil, 10 parts and 10 parts of vitamin E of DHA are uniformly mixed soft capsule content is made
Material, soft capsule content material and soft capsule shell material are then carried out the processing of pressure capsule on press device, and that black fruit fructus lycii seed oil is made is soft
Capsule.
Embodiment 2
A kind of application of the black fruit fructus lycii seed oil extract of the present invention in blood lipid-lowering medicine.
(1) description of test
Method used in following experiments is conventional method unless otherwise specified.
Material therefor, reagent etc., are commercially available unless otherwise specified in following experiments.
The specific side of producing of black fruit fructus lycii seed oil extract fatty acid and sterol of the present invention used in following experiments
Method are as follows: the black fruit fructus lycii seed after drying is crushed into weighing, then charging, (Nantong Huaan is super to be faced in supercritical extraction unit
Boundary extracts Co., Ltd, HA221-50-06 type) extraction kettle in, set postcritical extraction temperature as 40 DEG C, extracting pressure is
35 MPa, carbon dioxide flow are 40 kg/h, and extraction time is 90 min.
(2) experimental material
Test medicine: black fruit fructus lycii seed oil.Animal: zebra fish；KM mouse.Reagent: methylcellulose (Sigma, China)；It is different
Propyl alcohol (examination of Shanghai Shanghai, Chinese Shanghai, lot number 80109218)；Dimethyl sulfoxide/DMSO(Sigma-Aldrich, in China
Sea, lot number BCBT0803), TG, TC, LDL, HDL kit (Nanjing of China builds up Bioengineering Research Institute).
(3) laboratory apparatus
Desk type constant-temperatureoscillator oscillator (Chinese Shanghai Jing Hong experimental facilities Co., Ltd THZ-312), microplate reader (German Siemens
Healthineers)；High speed freezing centrifuge (German SIGMA 3-18K type), disecting microscope (SZX7, OLYMPUS,
Japan)；The camera (VertA1) being connected with microscope；(AZ100, Nikon are public for Power focus continuous zoom fluorescence microscope
Department)；Heartbeat blood flow analysis system (Zebralab3.3 (PB2084C))；Precision electronic balance (CP214, OHAUS,
America)；96 orifice plates are neglected and six orifice plates (Fisher Scientific, China)；
(4) efficacy validation is tested
1. black fruit fructus lycii seed oil extract fatty acid and sterol study zebra fish hypolipidemic activity
The translucent Albino strain zebra fish of 5 days melanin allelic variants of after fertilization is randomly selected in beaker, uses egg
Bloom feeding zebra fish establishes high blood lipid model, after model foundation, randomly selects 90 tail model zebra fish in six orifice plates, every hole
(experimental group) handles 30 tail zebra fish.DMSO hydrotropy gives black fruit fructus lycii seed oil, and concentration 0.1%, positive control drug Lip river is cut down
0.2 μM of concentration of statin, while model control group is set, every hole (experimental group) capacity is 3 mL.After test sample handles 48 h, use
Oil red O is dyed, and each experiment (concentration) group randomly selects 10 tail zebra fish and takes pictures and adopt under a dissecting microscope after dyeing
Collect data, analysis statistics zebra fish tail veins blood lipid optical density summation is commented according to the results of statistical analysis of optical density summation
The effect of valence black fruit fructus lycii seed oil in decreasing blood sugar.
As the result is shown: black fruit fructus lycii seed oil has drop to the zebra fish hyperlipidemia that yolk powder induces under 0.1% concentration conditions
Blood fat (is detailed in attached drawing 1 and table 1).
Experimental summary opinion: zebra fish activity research finds that black fruit fructus lycii seed oil has effect for reducing blood fat, therefore carries out to it
Activity research in Mice Body.
Zebra fish reducing blood lipid analysis result (n=10) after the processing of each test sample of table 1
2. black fruit fructus lycii seed oil extract fatty acid and sterol study hypolipidemic activity in Mice Body
Mouse high blood lipid model preparation: high lipid food Research Diets (D12079B) feeding modeling.
Experiment uses KM mouse 48, random 6 groups, every group 10, feeds different diet, respectively Normal group: common
Feed；High in fat group: high lipid food；Positive control drug group: high lipid food adds stomach-filling atorvastatin (0.125 mg/kg)；
High dose: high lipid food adds stomach-filling black fruit fructus lycii seed oily (15 g/kg)；Middle dosage: high lipid food adds stomach-filling black fruit fructus lycii seed oily
(10 g/kg)；Low dosage: high lipid food adds stomach-filling black fruit fructus lycii seed oily (5 g/kg)；Mouse freely ingests, drinks water, and record is each
The daily food ration of group mouse, food utilization (%)=(body weight evolution/total food ration) × 100 are continuous to feed 7 weeks.Daily
12 h of light application time, temperature control is at 20 ± 2 DEG C, humidity 50 ± 5%.After mouse is fed 7 weeks, 12 h of fasting weighs,
It plucks eyeball and takes blood, put to death and open mouse peritoneal and thoracic cavity, be immediately disconnected liver, heart, spleen, kidney, stomach fat and perirenal fat,
It weighs.It is to be centrifuged the time to obtain serum by one section of blood stasis, with TG, TC, LDL, HDL kit measurement blood lipid level.
The experimental results showed that (being detailed in attached drawing 2-5), the results show that black fruit fructus lycii seed oil can significantly reduce mouse high in fat
TC, TG, LDL are horizontal, raising HDL levels.Therefore, black fruit fructus lycii seed oil extract fatty acid and sterol can be used for preparing reducing blood lipid
Drug.
3. black fruit fructus lycii seed oil is studied with hyperlipidemia related target pancreatic lipase and cholesteryl ester enzyme inhibition activity
Pancreatic lipase inhibitory activity: pancreas rouge is prepared by the way that lipase to be dissolved in 100 mM Tris-HCl buffers (pH8.0)
Fat enzyme solutions (2.5 mg/ml), the buffer include 0.1% gum arabic and 0.2% NaTDC.By above-mentioned fat
Enzyme solutions are with 6000 rpm centrifugation 15 minutes to remove insoluble component.4-NPP is dissolved in isopropanol, distilled water is then used
It is diluted to required concentration.The final isopropyl alcohol concentration in solution is tested less than 1.0%.By the black fruit fructus lycii seed oil of various concentration
(1.28,2.56,4.80,6.40,9.60 and 12.80 mg/mL) and 80 μ L pancreatic lipases are added to 96 hole microtest plates
In and 37 DEG C incubate 10 minutes.Then the 4-NPP(1.0 mM of 80 μ L) is added in said mixture to start to react.It will
96 orifice plates incubate 30 minutes again at 37 DEG C, are then measured using microplate reader (Siemens Healthineers, Germany) anti-
Answer absorbance of the mixture at 405nm.Each measurement is triplicate to be carried out.It calculates suppression percentage and IC50 value (is detailed in attached
Fig. 6).
As oil concentration is in dose dependent increase, IC50 value is the pancreatic lipase inhibitory activity of black fruit fructus lycii seed oil oil
12.38±0.07 mg/mL。
Cholesteryl ester enzyme inhibition activity and mechanism: by the way that cholesterol esterase (CEase) is dissolved in 0.1M sodium phosphate buffer
2 μ g/mL CEase solution (0.03 U/mL) are prepared in liquid (pH 7.04), the buffer includes 0.1M NaCl.The bottom of enzyme
Object pNPB is dissolved in acetonitrile, tests the final acetonitrile concentration of solution less than 2.0%.Firstly, by the black fruit fructus lycii of various concentration
Seed oil and CEase be added in 96 hole microtest plates and 37 DEG C preincubate 5 minutes.It then, will be in pNPB(0.2 mM) addition
It states to start to react in mixture, and measures the absorbance of reaction mixture at 405 nm.Each experiment is triplicate to be carried out.
Calculate suppression percentage and IC50 value (being detailed in attached drawing 7).
Black fruit fructus lycii seed oil has dose-dependent inhibition effect to CEase, has stronger effective inhibiting effect, IC50
Value is 2.63 ± 0.13 mg/mL.In addition, research and dynamic analysis is inhibited to show inhibition of the black fruit fructus lycii seed oil to CEase
Mode belongs to the Noncompetition inhibition (being detailed in attached drawing 8-9) to reversible inhibition.
The foregoing is only a preferred embodiment of the present invention, is not intended to restrict the invention, for the skill of this field
For art personnel, the invention may be variously modified and varied.It is all within the principle of the present invention, it is made it is any modification, etc.
With replacement, improvement etc., should all be included in the protection scope of the present invention.
Claims (4)
1. a kind of application of black fruit fructus lycii seed oil extract in blood lipid-lowering medicine, which is characterized in that the black fruit fructus lycii seed
Oil extract is applied in blood lipid-lowering medicine as effective component.
2. a kind of application of the black fruit fructus lycii seed oil extract according to claim 1 in blood lipid-lowering medicine, feature exist
In, the black fruit fructus lycii seed oil extract can be used as effective component according to a conventional method with pharmaceutically acceptable any carrier system
At reducing blood lipid pharmaceutical formulation.
3. a kind of application of the black fruit fructus lycii seed oil extract according to claim 1 or 2 in blood lipid-lowering medicine, feature
It is, the fruit Boxthorn Seed Oil extract is fatty acid and sterol.
4. a kind of application of the black fruit fructus lycii seed oil extract according to claim 3 in blood lipid-lowering medicine, feature exist
In the fruit Boxthorn Seed Oil extract fat acid and sterol pass through supercritical fluid extraction.
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| CN114432364A (en) * | 2022-03-07 | 2022-05-06 | 中国科学院西北高原生物研究所 | Anti-inflammatory and blood fat-reducing composition, product, preparation method and application thereof |
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