CN110368392A - Corosolic acid is preparing the application in anti-liver injury medicament or anti-liver injury health care product - Google Patents
Corosolic acid is preparing the application in anti-liver injury medicament or anti-liver injury health care product Download PDFInfo
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- CN110368392A CN110368392A CN201910770488.9A CN201910770488A CN110368392A CN 110368392 A CN110368392 A CN 110368392A CN 201910770488 A CN201910770488 A CN 201910770488A CN 110368392 A CN110368392 A CN 110368392A
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- liver
- corosolic acid
- liver injury
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
-
- 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
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Gastroenterology & Hepatology (AREA)
- Virology (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Epidemiology (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention discloses Corosolic acids to prepare the application in anti-liver injury medicament or anti-liver injury health care product, belongs to Medicines and Health Product field.Corosolic acid (the Corosolic β-dihydroxyurs-12-en-28-oic of acid, 2 α, 3 acid, CA) also known as 2α-Hydroxyursolic Acid, molecular formula C30H48O4.Belong to α-botany bar gum alcohol type or Ursane pentacyclic triterpenoid, is primarily present in the plants such as loguat leaf, Lagerstroemia speciosa, fructus schisandrae, raspberry, valvate actinidia root, Leaves of Hippophae L, wild cherry.Corosolic acid of the invention has significant anti-liver injury effect, can be used for preparing prevention and/or prevents and treats the drug and health care product of anti-liver injury.
Description
Technical field
The invention belongs to Medicines and Health Product fields, and in particular to Corosolic acid is preparing anti-liver injury medicament or anti-liver injury
Application in health care product.
Background technique
Liver plays very important effect as intraperitoneal maximum organ in organism metabolism.Alcohol, drug, disease
The pathogenic factors such as poison, environmental factor and living habit can make liver damage, be destroyed so as to cause liver morphology structure and
The exception of liver function.
When dysfunction of liver, apparent metabolism obstacle, function of detoxification can be caused to reduce, the formation of bile and excretion hinder
Hinder.
It is estimated that there are about 400,000,000 people by all kinds of hepatopaths in the whole nation, in which: patients with nonalcoholic fatty liver disease is about 200,000,000
Example, accounts for 50.0% or so；Patients with alcoholic liver disease accounts for 15.0% or so there are about 60,000,000；Chronic hepatitis B, hepatitis C
Chronic viral hepatitis accounts for 26% there are about 1.4 hundred million, other liver diseases account for about 9.5%.It is serious that liver diseases have become China
Social public health problem.
Nonalcoholic fatty liver (non-alcoholic fatty liver disease, NAFLD) refers to except alcohol and its
He explicitly damages the clinical pathology syndrome that fatty over-deposit in liver cell caused by liver factor is main feature, is that the whole world is most normal
The liver diseases seen.Estimate according to meta-analysis, NAFLD is 25.24% in global illness rate.In China, as people are raw
The flat raising of running water and living habit, the change of dietary structure, its illness rate of China's last decade also increase substantially, up to 26%~
45%.
At present no matter traditional Chinese medicine or western modern medicine treatment NAFLD there is no specific medicament, have both improvement be metabolized and anti-inflammatory effect
Insulin sensitizer melbine and the Thiazolidinediones most advantage in pharmacological mechanism, but due to its hepatotoxicity wind agitation,
Weight the specific side effect such as increases and limits its extensive use clinically.
Corosolic acid also known as 2α-Hydroxyursolic Acid, molecular formula are as follows: C30H48O4 belongs to α-botany bar gum alcohol type or Ursane
Pentacyclic triterpenoid.Its structural formula is as follows:
It is primarily present in loguat leaf, Lagerstroemia speciosa, fructus schisandrae, raspberry, valvate actinidia root, Leaves of Hippophae L, wild cherry etc.
In plant.According to the literature, Corosolic acid has the injury of kidney of anti-diabetic and its induction, antitumor, anti-inflammatory, anti-cardiac muscle damage
Wound, antibacterial, inhibits the pharmacological actions such as osteoclast generates and bone dissolves at blood pressure lowering.
Summary of the invention
The present invention provides Corosolic acids to prepare the application in anti-liver injury medicament or anti-liver injury health care product, the liver
Damage is non-alcoholic hepatic injury, and the non-alcoholic hepatic injury is that simple fatty liver, steatohepatitis and fatty liver are fine
Dimensionization.
Further, in above-mentioned technical proposal, the hepatic injury further includes liver caused by alcoholic liver injury, virus infection
Damage, drug induced hepatic injury.
Further, in above-mentioned technical proposal, the non-alcoholic hepatic injury further includes cirrhosis and hepatocellular carcinoma.
Further, in above-mentioned technical proposal, the drug or health care product dosage form include oral agents and non-oral dose.
Further, in above-mentioned technical proposal, the oral agents include capsule, powder, pill, tablet, oral solutions.
Further, in above-mentioned technical proposal, described non-oral dose includes injection.
Corosolic acid of the invention can be made to treat and/or prevent the drug or health care product of anti-liver injury, described
Drug or health care product liquid, solid, semisolid is made, take orally or it is non-oral, capsule, powder, pill, tablet, oral solution,
Pharmaceutically acceptable any dosage form such as injection.
Simple fatty liver of the present invention is the early reaction of nonalcoholic fatty liver, in liver fatty synthesis with point
Solution metabolism disequilibrium, or transports generation obstacle, fat will in liver cell excess accumulation.Fatty excess accumulation will result in
The duration of liver damages, and leads to the steatosis of liver cell, the disorder of lipid-metabolism, makes liver to inflammatory reaction and various livers
The neurological susceptibility of impairment factor increases, and then promotes the generation and development of hepatic fibrosis-renal tubular ectasia syndrome.Nonalcoholic fatty liver disease is with liver
The liver diseases that dirty fatty infiltration, inflammatory reaction, hepatocellular damage, necrosis and fibrosis are characterized.Non-alcoholic fatty liver
If scorching diagnosing and treating not in time, can progress to liver fibrosis, cirrhosis, portal hypertension, hepatic failure and hepatocellular carcinoma,
Therefore, it treats ahead of time increasingly important.
Drug of the invention is the effective component of various plants, from a wealth of sources, has significant curative effect to anti-liver injury.This
Invention discovery Corosolic acid has effects that anti-liver injury.Corosolic acid can improve to conspicuousness 60%kcal high lipid food and lure
The C57BL/6j fatty liver mice serum and aspartate transaminase (AST) led are horizontal and liver fat is accumulated, to pure
Fatty liver has good therapeutic effect；The acute hyperlipemia that Corosolic acid can improve to conspicuousness tyloxapol induction is simultaneous
Have fatty liver ICR mouse ALT, AST, triglycerides and cholesterol levels, liver fat accumulation and inflammation；Corosolic acid can
Improve to conspicuousness 60%kcal high lipid food and carbon tetrachloride combined induction has both fatty liver inflammation and fatty liver is fine
The C57BL/6j mouse adipose liver inflammation and fatty liver fibrosis of dimensionization.
Detailed description of the invention
Fig. 1 is the mouse blood aspartate transaminase that 1 Corosolic acid of embodiment induces 60%kcal high lipid food
It influences.
Fig. 2 be the C57BL/6j mouse liver tissue H&E that induce to 60%kcal high lipid food of 1 Corosolic acid of embodiment with
Oil red O dyeing.
Fig. 3 is that the mouse alanine aminotransferase (figure A) that 2 Corosolic acid of embodiment induces tyloxapol and aspartic acid turn
The influence of adnosine deaminase (figure B), triglycerides (figure C) and cholesterol (figure D) level.
Fig. 4 is that 2 Corosolic acid of embodiment dyes ICR mouse liver H&E and Oil the red O that tyloxapol induces.
Fig. 5 is ICR mouse liver F4/80 and the caspase-1 expression that 2 Corosolic acid of embodiment induces tyloxapol
It influences.
Fig. 6 is 3 Corosolic acid of embodiment to high lipid food-CCl4Mouse overall appearance (figure A), the fresh liver of induction
The influence of (figure B), liver index (figure C) and blood alanine aminotransferase and aspartate transaminase level (figure D).
Fig. 7 is 3 Corosolic acid of embodiment to high lipid food-CCl4The mouse liver histopathological analysis of induction.
Fig. 8 is 3 Corosolic acid of embodiment to high lipid food-CCl4The mouse liver tissue IL-1 β, F4/80 of induction and
The influence of caspase-1 expression.
Specific embodiment
Following nonlimiting examples can with a person of ordinary skill in the art will more fully understand the present invention, but not with
Any mode limits the present invention.
Material and reagent involved in the embodiment of the present invention are unless otherwise instructed commercial product.
1 Corosolic acid of embodiment improves the simple fatty liver mouse liver fat deposition of 60%kcal high lipid food induction
Effect
(1) experimental animal: 5 week old C57BL/6j mouse freely ingest and drink water, room temperature (23 ± 2 DEG C), natural lighting.
(2) experimental method:
A. mouse mice group: is randomly divided into 5 groups: normal group (CON), model group (HFD), Corosolic acid low dose group
(CAL), Corosolic acid high dose group (CAH), positive controls (Met).
B. feeding method: (H10010, Beijing China Fukang biotechnology share are limited with 10%kcal chow diet for normal group
Company) it feeds, remaining four groups are used 60%kcal high lipid food (H10060, Beijing HFK Bio-Technology Co., Ltd.)
It feeds, feeds 8 weeks in total.
C. the measuring method of administration and physiochemical indice: latter 4 weeks, the drug of corresponding dosage was given in stomach-filling to groups of animals respectively,
Drug is dissolved in 0.5% sodium carboxymethylcellulose (CMC) solution, and the 0.5%CMC that normal group and model group give respective volume is molten
Liquid, Corosolic acid low dose group and Corosolic acid high dose group give the Corosolic acid (CA) of 10mg/kg and 20mg/kg respectively,
Positive controls give the melbine of 300mg/kg, once a day.Mice plasma is acquired using retroorbital venous clump blood collection method,
At 4 DEG C, 3000rpm, it is centrifuged 20min, supernatant is taken to prepare serum.Aspartate transaminase is measured by the operating instruction of kit
(AST) horizontal, aspartate transaminase kit (Bioengineering Research Institute is built up in C010-2-1, Nanjing).
D. the preparation of frozen section: it is put into a small amount of O.C.T. frozen embedding liquid on mold, tissue block is adjusted to be convenient for
The position of slice, lies against in mold, then adds O.C.T. frozen embedding liquid to submerge tissue in right amount, and mold is put into cryostat
Middle frost is blocking.Frost is completed to be placed on cryostat frozen section.5 μm of frozen section.Frozen section is attached to glass slide
Afterwards, it is immediately placed in cryostat and saves.
E.H&E dyeing: the liver cut, histotomy are put into ether/ethanol (1:1) fixer and fix 1min, water
It washes 1min, haematoxylin dyeing 1min, washes 3min, eosin stains liver organization 2min, wash 1min, graded ethanol (70%,
80%, 90%, 95%) dehydration, dimethylbenzene is transparent, neutral gum mounting.Microscopically observation cellular morphology is simultaneously taken pictures.
F. oil red O stain: the liver tissue slices cut being put into 10% neutral formalin fixer and fix 5min, is washed
30s, 60% isopropanol 5min, oil red O- aqueous isopropanol tune dye 3min, wash 30s, haematoxylin dyeing 3s, wash 5min,
Glycerin gelatine mounting.
(3) experimental result:
As shown in Figure 1, normally group is compared with model group, p < 0.001 * * *；Model group compared with Corosolic acid administration group,###
p<0.001。
The experimental results showed that model group mouse aspartate transaminase (AST) is horizontal significantly raised, p compared with normal group
<0.001；Compared with model group, Corosolic acid administration group and positive controls AST level are significantly reduced, p < 0.001.
From figure 2 it can be seen that normally without finding apparent fat drips, model group mouse liver group in the liver of group mouse
It can be observed many fat drips being dispersed in knitting, and fat drips in Corosolic acid administration group and melbine positive group mouse liver tissue
Quantity it is obviously fewer than model group, and in high dose administration group liver fat drips quantity be less than melbine administration group.
To sum up, the results showed that, Corosolic acid can improve to conspicuousness the C57BL/6j of 60%kcal high lipid food induction
Fatty liver mice serum and aspartate transaminase (AST) level and liver fat accumulation, have simple fatty liver good
Good therapeutic effect.
2 Corosolic acid of embodiment has both the improvement result of fatty liver to the chmice acute hyperlipemia that tyloxapol induces
(1) experimental animal: the ICR male mice of 6 week old freely ingests and drinks water, room temperature (23 ± 2 DEG C), natural lighting.
(2) experimental method:
A. mouse mice group: is randomly divided into normal group (CON), Corosolic acid administration group (CA), model group (TY), section
Roseau acid low dose group (TCL), Corosolic acid high dose group (TCH) and positive controls (TS).
B. medication: after mouse fasting 12h, the sodium carboxymethylcellulose that normal group and model group give 0.5% is molten
Liquid, the Corosolic acid of Corosolic acid administration group (CA) stomach-filling 30mg/kg, section sieve of Corosolic acid low dose group stomach-filling 15mg/kg
Rope acid, the Corosolic acid of Corosolic acid high dose group stomach-filling 30mg/kg, the Simvastatin of positive controls stomach-filling 20mg/kg.1h
Afterwards, the tyloxapol of 500mg/kg is injected intraperitoneally in normal group intraperitoneal injection aqua sterilisa, other groups.
C. the measuring method of physiochemical indice: putting to death animal after 12h, acquires mouse blood using retroorbital venous clump blood collection method
Slurry is centrifuged 20min, supernatant is taken to prepare serum at 4 DEG C, 3000rpm.Serum measures third by the operating instruction of kit respectively
Propylhomoserin transaminase (ALT), aspartate transaminase (AST) triglycerides and cholesterol levels.The alanine aminotransferase reagent
Box (Bioengineering Research Institute is built up in C009-2-1, Nanjing), (life is built up in C010-2-1, Nanjing to aspartate transaminase kit
Object Graduate School of Engineering), triglyceride reagent box (Bioengineering Research Institute is built up in A110-1-1, Nanjing), Cholesterol Kit
(Bioengineering Research Institute is built up in A111-1-1, Nanjing).
D. the preparation of frozen section: it is put into a small amount of O.C.T. frozen embedding liquid on mold, tissue block is adjusted to be convenient for
The position of slice, lies against in mold, then adds O.C.T. frozen embedding liquid to submerge tissue in right amount, and mold is put into cryostat
Middle frost is blocking.Frost is completed to be placed on cryostat frozen section.5 μm of frozen section.Frozen section is attached to glass slide
Afterwards, it is immediately placed in cryostat and saves.
E.H&E dyeing: the liver cut, histotomy are put into ether/ethanol (1:1) fixer and fix 1min, water
It washes 1min, haematoxylin dyeing 1min, washes 3min, eosin stains liver organization 2min, wash 1min, graded ethanol (70%,
80%, 90%, 95%) dehydration, dimethylbenzene is transparent, neutral gum mounting.Microscopically observation cellular morphology is simultaneously taken pictures.
F. oil red O stain: the liver tissue slices cut being put into 10% neutral formalin fixer and fix 5min, is washed
30s, 60% isopropanol 5min, oil red O- aqueous isopropanol tune dye 3min, wash 30s, haematoxylin dyeing 3s, wash 5min,
Glycerin gelatine mounting.
G. immunohistochemistry: frozen section is fixed with acetone.Slice is put into 0.3% hydrogen peroxide methanol solution of people and impregnates 30s,
To eliminate activating oxide enzyme in tissue.Slice takes out to be put into distilled water and rinse 2 times, each 3min.PBS liquid rinsing 2 times, often
Secondary 5min.Microwave digestion further takes out slice and is placed in wet box, and 10% corresponding serum is added dropwise in every slice, places 37 DEG C, 30min.
Discard serum solution, then be added dropwise 3d corresponding antibodies [F4/80 antibody (sc-377009, Santacruz biotechnology) and
Caspase-1 antibody (sc-392736, Santacruz biotechnology)] 1:1000 dilution, place 4 DEG C overnight.Use PBS
Liquid rinse 3 times, each 5min, be added dropwise biotin labeling secondary antibody [mouse IgG kappa binding protein (sc-516102,
Santacruz biotechnology)], room temperature l h.Secondary antibody is discarded, PBS develops a film, and DAB developing solution is added dropwise on every slice
(k346711-2, dako), distilled water rinse 2 times, and haematoxylin is redyed, then row dehydration, copal gum mounting.
(3) experimental result:
As shown in figure 3, model group is compared with normal group, ###p < 0.001；Administration group is compared with model group, * * p <
0.01, * * * p < 0.001.After mouse peritoneal injects tyloxapol, ALT, AST, triglycerides and cholesterol levels are significantly risen
High (p < 0.001).Compared with model group, after giving Corosolic acid, Corosolic acid low dose group and Corosolic acid high dose group
ALT, AST, triglycerides and cholesterol levels dose-dependently reduce.
Figure 4, it is seen that without finding apparent fat drips, TY group mouse in CON group mouse and CA group mouse liver
Many fat drips being dispersed in can be observed in liver organization, and in Corosolic acid administration group mouse liver fat drips quantity dose-dependant
Reduce to property.
From figure 5 it can be seen that CON group mouse and CA group mouse liver do not show brown, TY group mouse liver tissue is aobvious
Show dark brown brown, and Corosolic acid administration group mouse liver sepia lighter.Illustrate that Corosolic acid induces tyloxapol
The expression of the inflammatory factors such as mouse liver F4/80 and caspase-1 have inhibiting effect.
To sum up, the results showed that, the acute hyperlipemia that Corosolic acid can improve to conspicuousness tyloxapol induction has both
Fatty liver ICR mouse ALT, AST, triglycerides and cholesterol levels, liver fat accumulation and inflammation.
C57BL/6j mouse adipose of 3 Corosolic acid of embodiment to 60%kcal high lipid food and carbon tetrachloride combined induction
Property hepatitis has both the improvement result of fatty liver fibrosis
(1) experimental animal: the C57BL/6j male mice of 5 week old, room temperature (23 ± 2 DEG C), natural lighting.
(2) experimental method:
A. mice group and medication: the free diet of mouse adapts to environment after a week, is randomly divided into 5 groups: normal group
(Control, CON), model group (High-fat diet+CCL4, HC)；Corosolic acid low dose group (High-fatdiet+CCL4
+ 10mg/kg CA, HCCL), Corosolic acid high dose group (High-fat diet+CCL4+ 20mg/kgCA, HCCH) and the positive it is right
According to group (High-fat diet+CCL4+ 30mg/kg Silymarin, HCS), every group 8.Normal group is raised is raised with 10%kcal
It is primary (0.2 μ l/g) that corn oil is injected intraperitoneally weekly in material；Other groups are raised with 60%kcal high lipid food, are injected intraperitoneally weekly
Carbon tetrachloride is primary (0.2 μ l/g), feeds 12 weeks altogether.10 weeks afterwards, the drug of the daily corresponding dosage of stomach-filling of mouse, normal group
With 0.5% sodium cellulose glycolate solution of model group stomach-filling corresponding dosage, Corosolic acid low dose group gives the section of 10mg/kg
Roseau acid；Corosolic acid high dose group gives the Corosolic acid of 20mg/kg；Positive controls give the silymarin of 30mg/kg.
Drug is dissolved in 0.5% sodium cellulose glycolate solution.
B. sample collection and physiochemical indice measuring method: mouse web portion is dissected, mouse liver appearance is observed；It takes out
Mouse liver carries out the calculating of liver index；Serum is taken to measure alanine aminotransferase (ALT) respectively by the operating instruction of kit
With aspartate transaminase (AST) level, the alanine aminotransferase kit (build up bioengineering and grind by C009-2-1, Nanjing
Study carefully institute), aspartate transaminase kit (Bioengineering Research Institute is built up in C010-2-1, Nanjing).
C. the preparation of frozen section: it is put into a small amount of O.C.T. frozen embedding liquid on mold, tissue block is adjusted to be convenient for
The position of slice, lies against in mold, then adds O.C.T. frozen embedding liquid to submerge tissue in right amount, and mold is put into cryostat
Middle frost is blocking.Frost is completed to be placed on cryostat frozen section.5 μm of frozen section.Frozen section is attached to glass slide
Afterwards, it is immediately placed in cryostat and saves.
D.H&E dyeing: the liver cut, histotomy are put into ether/ethanol (1:1) fixer and fix 1min, water
It washes 1min, haematoxylin dyeing 1min, washes 3min, eosin stains liver organization 2min, wash 1min, graded ethanol (70%,
80%, 90%, 95%) dehydration, dimethylbenzene is transparent, neutral gum mounting.Microscopically observation cellular morphology is simultaneously taken pictures.
E. oil red O stain: the liver tissue slices cut being put into 10% neutral formalin fixer and fix 5min, is washed
30s, 60% isopropanol 5min, oil red O- aqueous isopropanol tune dye 3min, wash 30s, haematoxylin dyeing 3s, wash 5min,
Glycerin gelatine mounting.
F. sirius red dyes: 0.05% sirius red solution of the liver tissue slices cut is dyed 30min, water
It washes 3min twice, dyes 2hr with saturation picric acid solution, washed 2 times, 3min with 0.01mol/L HCl solution, with graded ethanol point
Change and be dehydrated, dimethylbenzene is transparent, neutral gum mounting.
G.western-blot experiment: weighing liver organization in centrifuge tube, and 1ml protein lysate, tissue homogenate is added
Afterwards, 20min is centrifuged under 4 DEG C, 13000rpm, collecting supernatant is liver total protein.The quantitative of protein uses BCA method according to examination
Agent box specification (Bio-rad) carries out time-and-motion study.It is (every that a certain amount of 5x Loading Buffer is added in protein example
The 5x Loading Buffer of 10 μ L is added in 40 μ L protein), it is uniformly mixed, in 100 DEG C of heat denatureds, is centrifuged after cooling.
Electrophoresis is carried out after loading at 100v.It, will be on Protein transfer to pvdf membrane after electrophoresis.It is closed in 5% skimmed milk power
2h.After closing, primary antibody [IL-1 β antibody (ab9722, abcam), F4/80 antibody (sc-377009, Santacruz
Biotechnology) and caspase-1 antibody (sc-392736, Santacruz biotechnology)] be incubated overnight, two
It is anti-that [IL-1 β is to corresponding secondary antibody mouse anti-rabbit IgG-HRP (sc-2357)；F4/80 and caspase-1 are corresponding
Secondary antibody be mouse IgG kappa binding protein (sc-516102, Santacruz biotechnology)] incubation at room temperature 2h.
It is developed the color with ECL, after reacting 3min.In darkroom, developed with X-ray film.
(3) experimental result:
Mouse web portion is dissected, it is ruddy as can be seen that normal group mouse liver surface is smooth from Fig. 6 A and 6B,
High resilience, and model group mouse liver rough surface, there is granular sensation, without elasticity, liver volume slightly increases, and coating is nervous
It is smooth, edge rust, color Huang.Corosolic acid high dose group mouse liver appearance is similar with Normal group, restores red gloss,
Corosolic acid low dose group and positive controls also suitably improve the appearance of liver.
From Fig. 6 C as can be seen that compared with Normal group, model group mouse liver index apparent increase (p <
0.001)；Compared with model group, Corosolic acid administration group liver index is decreased significantly.Prompt high lipid food-CCl4Joint
Being administered can induce mouse liver injury, and Corosolic acid can improve high lipid food-CCl4The mouse liver injury of induction.
It can be seen that compared with Normal group from Fig. 6 D, alanine aminotransferase (ALT) in model group mice serum,
Aspartate transaminase (AST) is horizontal significantly raised (p < 0.001)；Compared with model group, Corosolic acid high dose group and Ke Luo
It is horizontal (p < 0.05) that rope acid low dose group significantly reduces Serum ALT, AST.
As can be seen that CON group liver tissues of rats has no significant change, liver cell bar rope row from the H&E coloration result of Fig. 7
It arranges neatly, has no obvious denaturation, necrosis, obvious inflammatory cell infiltration, major part in HC group liver tissues of rats are had no in liver parenchyma
Leaflet structure is destroyed, and the large stretch of necrosis of hepatic tissue is in abortive haul shape, and visible fat is denaturalized therebetween, it is seen that a large amount of inflammatory cell leachings
Profit, Corosolic acid low dose group focal necrosis visible with positive administration group murine liver tissue, there is a little inflammatory cell infiltration, rouge
Fat denaturation is lighter.
As can be seen that CON group mouse liver exists without obvious fat drips from the oil red O stain result of Fig. 7；With CON group ratio
Compared with HC group mouse liver is dispersed in very more fat drips；Compared with HC group, Corosolic acid administration group and positive controls Mouse Liver
Fat drips quantity in dirty has different degrees of reduction, and fat drips quantity is considerably less than positive controls in high dose group mouse liver,
And fat drips are less than low dosage administration group in high dose administration group mouse liver.
It can be seen that CON group hepatic tissue is normal from the sirius red coloration result of Fig. 7, HC group rat liver normal hepatocytes
Leaflet structure disappears, and with apparent fibrous connective tissue hyperplasia, forms typical pseudolobuli structure.And Corosolic acid administration group
The degree of fibrosis of mouse liver is significantly lower than model group.
From the immunoblot experiment result of Fig. 8 as can be seen that compared with CON group, HC group mouse liver tissue IL-1 β,
The protein expressions such as F4/80, caspase-1 obviously deepen, and these protein in Corosolic acid administration group mouse liver tissue
Expression obviously tail off.Illustrate that Corosolic acid combines CCl to 60%kcal high lipid food4Mouse liver IL-1 β of combined induction,
The expression of the inflammatory factors such as F4/80 and caspase1 has inhibiting effect.
To sum up, the results showed that Corosolic acid can improve to conspicuousness 60%kcal high lipid food and carbon tetrachloride is combined and lured
Lead C57BL/6j mouse adipose liver inflammation and the fatty liver fibre for having both fatty liver inflammation and fatty liver fibrosis
Dimensionization.
Claims (6)
1. Corosolic acid is preparing the application in anti-liver injury medicament or anti-liver injury health care product, which is characterized in that the liver damage
Wound is non-alcoholic hepatic injury, and the non-alcoholic hepatic injury is simple fatty liver, steatohepatitis and fatty liver fiber
Change.
2. application according to claim 1, which is characterized in that the hepatic injury further includes alcoholic liver injury, virus sense
Hepatic injury, drug induced hepatic injury caused by contaminating.
3. application according to claim 1, which is characterized in that the non-alcoholic hepatic injury further includes that cirrhosis and liver are thin
Born of the same parents' cancer.
4. application according to claim 1, which is characterized in that the drug or health care product dosage form include oral agents and non-mouth
Take agent.
5. application according to claim 4, which is characterized in that the oral agents include capsule, powder, pill, tablet
And oral solutions.
6. application according to claim 4, which is characterized in that described non-oral dose includes injection.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113841816A (en) * | 2021-09-18 | 2021-12-28 | 中国水产科学研究院长江水产研究所 | Application of corosolic acid in preparation of feed for preventing and/or treating fish fatty liver |
| CN113841816B (en) * | 2021-09-18 | 2024-04-02 | 中国水产科学研究院长江水产研究所 | Use of corosolic acid as fish feed additive for altering fatty acid composition of fish liver lipid |
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