CN107349252A - Psoralea corylifolia is applied in the medicine for preparing treatment children's fatty liver - Google Patents

Psoralea corylifolia is applied in the medicine for preparing treatment children's fatty liver Download PDF

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CN107349252A
CN107349252A CN201710419208.0A CN201710419208A CN107349252A CN 107349252 A CN107349252 A CN 107349252A CN 201710419208 A CN201710419208 A CN 201710419208A CN 107349252 A CN107349252 A CN 107349252A
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psoralea corylifolia
fatty liver
weight
children
group
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周俪姗
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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)
    • A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
    • A61K36/487—Psoralea
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
    • A01K67/00—Rearing or breeding animals, not otherwise provided for; New breeds of animals
    • A01K67/02—Breeding vertebrates
    • 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)
    • A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/16—Agglomerates; Granulates; Microbeadlets ; Microspheres; Pellets; Solid products obtained by spray drying, spray freeze drying, spray congealing,(multiple) emulsion solvent evaporation or extraction
    • A61K9/1605—Excipients; Inactive ingredients
    • A61K9/1629—Organic macromolecular compounds
    • A61K9/1652—Polysaccharides, e.g. alginate, cellulose derivatives; Cyclodextrin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/20—Pills, tablets, discs, rods
    • A61K9/2004—Excipients; Inactive ingredients
    • A61K9/2022—Organic macromolecular compounds
    • A61K9/205—Polysaccharides, e.g. alginate, gums; Cyclodextrin
    • A61K9/2059—Starch, including chemically or physically modified derivatives; Amylose; Amylopectin; Dextrin

Abstract

The present invention relates to pharmaceutical technology field, is applied in particular to a kind of psoralea corylifolia in the medicine for treating children's fatty liver is prepared.Psoralea corylifolia can improve young mice fatty liver, and curative effect is in dose dependent；Improve young age fatty liver mouse Early hepatic fibrosis, curative effect is in dose dependent；The activation of NF κ B Inflammatory Pathways can be suppressed, curative effect is in dose dependent.The present invention proves that psoralea corylifolia has the function that to suppress hepatic tissue NF kB activities by establishing young mice NAFLD models, so as to have preventive and therapeutic action to children's fatty liver.Psoralea corylifolia can also be shared with other compound groups in the pharmaceutical preparation for preparing treatment children's fatty liver.

Description

Psoralea corylifolia is applied in the medicine for preparing treatment children's fatty liver
Technical field
The present invention relates to pharmaceutical technology field, in particular to a kind of psoralea corylifolia in the medicine for treating children's fatty liver is prepared Using.
Background technology
Non-alcohol fatty liver (nonalcoholic fatty liver disease, NAFLD) presentation globalization, Become younger fashion trend, is the primary cause of disease of childhood visits digestion training or hepatopathy training.It is fat in close relations with fatty liver, I The prevalence of state's childhood obesity has been objective fact, and Obese children NAFLD illness rates are up to 68.2%.Originating from childhood NAFLD is even more be grown up hepatic sclerosis, the main standby crowd of metabolic syndrome related component.With Urbanization in China, middle production Family emerges and the all-round opening of " two tires " policy, NAFLD spread rapidly, substituted chronic viral hepatitis, turned into and threaten me The significant problem of state's children's health.
At present, clinical anti-Adult Fatty Liver medicine lacks such as, just more lacks for children NAFLD active drug.However, It is different from adult, children's nonalcoholic fatty liver disease (nonalcoholic steatohepatitis, NASH) portal area disease It is generally serious compared with leaflet to become (inflammation and fibrosis).In recent years, " Nuclear factor kappa B (nuclear factor κ B, NF- κ B) " phase Closing critical role of the Inflammatory Pathway in NAFLD pathogenesis is constantly confirmed.
Effects of Bu Gu fat (Psoralea corylifolia L., PC) is traditional common medicine of paediatrics, has warm spleen kidney tonifying The medicinal feature of establishing-Yang.The modern pharmacology research of the medicine is mainly related to adult disease, and psoralea corylifolia has treatment tumour and changed Kind osteoporosis and other effects, but at present, therapeutic actions and its Mechanism Study of the PC to children's fatty liver have no report.
In addition to antitumor and improve osteoporosis and other effects, still, therapeutic actions and its mechanism of the PC to children's fatty liver Research has no report.
The content of the invention
Applied it is an object of the present invention to provide a kind of psoralea corylifolia in the medicine for preparing treatment children's fatty liver.
To achieve the above object, a kind of psoralea corylifolia provided by the invention should in the medicine for preparing treatment children's fatty liver With.
Preferably, children's fatty liver is children's nonalcoholic fatty liver disease NASH.
Present invention also offers a kind of pharmaceutical preparation for treating children's fatty liver, the psoralea corylifolia containing effective dose and pharmaceutically Acceptable auxiliary material and/or auxiliary material.
Psoralea corylifolia can also be shared with other compound groups in prepare treatment children's fatty liver pharmaceutical preparation, particularly with Fenugreek combination prepares the pharmaceutical preparation for being used for treating children's fatty liver.
Further, the auxiliary material is vitamin C, sorbierite, mannitol, xylitol, fructose, amino acid, meglumine, paste Any one in essence, magnesium stearate and sucrose or two kinds.
Pharmaceutical preparation of the present invention can select different dosage forms according to route of administration, and the pharmaceutical preparation is oral formulations or filling Enteral formulationses.
Preferably, the oral formulations are tablet, oral liquid or granule.
Preferably, the oral formulations are by the psoralea corylifolia of 1 parts by weight, the vitamin C of 3 parts by weight and 1 parts by weight of sucrose.
Preferably, the tablet is by the psoralea corylifolia of 1 parts by weight, the fenugreek of 1 parts by weight, the sucrose of 3 parts by weight and 1 weight The dextrin of part.
Preferably, the granule by the psoralea corylifolia of 1 parts by weight, the fenugreek of 1 parts by weight, 3 parts by weight sucrose and 1 weight Measure the dextrin of part.
Present invention discover that psoralea corylifolia has following pharmacological action：
1) psoralea corylifolia has prevention and treatment to children's fatty liver (especially children nonalcoholic fatty liver disease NASH) Effect
The present invention is by establishing young mice NAFLD models, detection blood glucose, blood fat (TC, TG, LDL-C, HDL-C), empty stomach Insulin, liver function (ALT, AST) are horizontal, calculate insulin resistance index (HOMA-IR), and observation liver morphology changes, Liver tg (TG) content, CD44 protein expressions are detected, while detects hepatic tissue Nuclear factor kappa B (NF- κ B) p65 phosphorylations (p-p65) and non-phosphorylating albumen (p65) is expressed and its downstream cytokine (TNF-α, IL-8) is horizontal, it was demonstrated that psoralea corylifolia has Suppress the effect of hepatic tissue NF- kB activities, so as to children's fatty liver (especially children nonalcoholic fatty liver disease NASH) With preventive and therapeutic action.
2) PC can improve young mice fatty liver, and curative effect is in dose dependent.
The method that the present invention uses High-fat diet, can be successfully established young mice Models of Fatty Liver.The model mice Drawn materials before body maturation (10 week old), occur insulin resistance, liver function damage (ALT ＞ AST), liver before adult TG contents increase, liver histological shows the performance such as steatosis and cell infiltration, possess NAFLD characteristic features.Give and mend After bone fat particle gavage, model mice HOMA-IR is reduced, ALT and the horizontal reductions of AST, liver TG contents are reduced, and high dose group is excellent In low dose group.Further contrast liver pathology and change hepatic cell fattydegeneration mitigation, cell infiltration after display PC treatments Reduce, and high dose group is better than low dose group.Illustrate that PC can improve young mice fatty liver, curative effect is in dose dependent.
3) PC can improve young age fatty liver mouse Early hepatic fibrosis, and curative effect is in dose dependent.
The present invention is by have detected hepatic tissue CD44 protein expressions.CD44 is I type transmembrane proteins, is cell surface receptor, It is often expressed as in liver Kupffer cells and lymphocyte infiltration surface., can be with overexpression when fibre modification occurs in liver Extracellular matrix product hyaluronic acid is specifically bound.Therefore, CD44 is considered as assessing the indirect indexes of Early hepatic fibrosis. Portal area proliferation of fibrous tissue is shown in model mice liver HE dyeing, and the protein expression of portal area lymphocyte CD 44 is in strong positive, Liver fibrosis is prompted to exist.After PC is treated, observation liver HE dyeing, low dose group mouse portal area fibroplasia mitigates, High dose group disappears；Hepatic tissue CD44 protein expressions decline, and high dose group relatively low-dose group, which declines, to be become apparent from.Illustrate that PC can improve Young age fatty liver mouse Early hepatic fibrosis, curative effect are also in dose dependent.
4) PC can suppress the activation of NF- κ B Inflammatory Pathways, and curative effect is in dose dependent.
NAFLD morbidity is related to multiple mechanism such as oxidative stress, insulin resistance and inflammation, and mutual shadow between each mechanism Ring and promote mutually.Chronic inflammatory reaction is not only played an important role in above-mentioned association, also promotes liver fibrosis process.NF-κB Inflammatory signals Pathway Activation is proved that also display model is small by the present invention in numerous clinics about NAFLD and laboratory research Rat liver phosphorylation and non-phosphorylating NF- κ B p65 protein expression ratios are significantly raised, and with NF- κ B downstreams inflammatory factor Hepatic tissue TNF-α, IL-8 expression increase.PC treatment groups are further detected, are found：Model mice is after PC is treated, hepatic tissue phosphorus Acidifying is decreased obviously with non-phosphorylating NF- κ B p65 protein expression ratios, and hepatic tissue TNF-α, IL-8 levels also significantly reduce. Illustrate that the effect of PC improves fatty liver may be by suppressing the activation of NF- κ B Inflammatory Pathways to realize.Meanwhile high dose group It is obvious to lower intensity relatively low-dose group.
Brief description of the drawings
Fig. 1 is that each group mouse liver histopathological findings compare figure (HE is dyed, × 400)；
In figure, A：Normal group；B：Model group；C：Psoralea corylifolia (low dosage) group；
D：Psoralea corylifolia (high dose) group；E：Vitamin E group；
Fig. 2 is that (× 200, arrow show portal area lymphocyte sun to each group mouse liver tissue CD44 expressions figure Property expression CD44)；
In figure, A：Normal group；B：Model group；C：Psoralea corylifolia (low dosage) group；D：Psoralea corylifolia (high dose) group；E：Vitamin E groups；
Fig. 3 be each group murine liver tissue p-p65/p65 odds ratios relatively scheme (N=3)；
In figure, N：Normal group；M：Model group；PC(L)：Psoralea corylifolia (low dosage) group；PC(H)：Psoralea corylifolia (high dose) group； VitE：Vitamin E group is (compared with normal group▲▲p<0.01；Compared with model group△△p<0.01；With psoralea corylifolia low dose group ratio Compared with * p<0.05).
Embodiment
In order to preferably explain the present invention, below in conjunction with the specific embodiment main contents that the present invention is furture elucidated, but Present disclosure is not limited solely to following examples.
The experimental animal and material that the present invention uses are as follows：
1. experimental animal
SPF levels male C 57 BL/6 J mouse (3 week old) 40, purchased from Beijing HFK Bio-Technology Co., Ltd. (Quality of Experimental Animals quality certification number：NO.11401300041293).
2. feed
D12451 high lipid foods are purchased from Beijing HFK Bio-Technology Co., Ltd., are formulated and are：45%kcal fat, 20%kcal protein, 35%kcal carbohydrate.Normal diet is by Tongji Medical College, Huazhong Science and Technology Univ. experimental animal The heart provides, and is formulated and is：35% flour, 20% soy meal, 20% corn flour, 15.5% wheat bran, 0.5% soya-bean oil, 5% fish meal, 2.5% bone powder, 1% dusty yeast, 0.5% salt.
3. medicine and main agents
Psoralea corylifolia granule provides (batch number by China Resources Sanjiu Medical ＆ Pharmaceutical Co., Ltd.：1601002S).
Vitamin E soft capsule is purchased from Zhejiang Medicine Co.
Blood glucose (BG) test paper, Cocktail protease inhibitors are purchased from Roche companies of Switzerland.
Glucose estimation kit is purchased from Shanghai famous classic bioengineering Co., Ltd.
T-CHOL (TC), triglycerides (TG), HDL-C (HDL-C), low-density lipoprotein courage are consolidated Alcohol (LDL-C) testing cassete；Glutamic-pyruvic transaminase (ALT), glutamic-oxalacetic transaminease (AST) kit；Mouse interleukin -8 (IL-8) Enzyme-linked immunologic detecting kit is purchased from Nanjing and builds up Science and Technology Ltd..
Mouse islets element (INS) enzyme-linked immunosorbent assay kit, murine tumor necrosis factor α (TNF-α) is enzyme-linked exempts from Epidemic disease determining adsorption kit is purchased from Wuhan Yi Lairuite bio tech ltd.
EDTA repairs liquid, BCA determination of protein concentration kit, RIPA total proteins lysate, PMSF, the suppression of phosphorylated protein enzyme Preparation is purchased from Wuhan ASPEN Bioisystech Co., Ltd.DAB colour reagents box is limited purchased from Beijing Zhong Shan Golden Bridge biotechnology Company.
Rabbit anti-mouse CD44 monoclonal antibodies, GAPDH polyclonal antibodies, phosphorylation NF- κ B p65 (phosphorylation sites S276) polyclonal antibody is purchased from Abcam companies.Rabbit anti-mouse NF- κ B p65 monoclonal antibodies are purchased from CST companies.Goat resists Rabbit polyclonal secondary antibody is purchased from KPL companies.
4. model is prepared, is grouped and handled
Mouse adaptability is fed 3 days.It is normal group to randomly select 8 mouse and compile, and continues chow diet and feeds.Remaining 32 It is 4 groups that mouse is compiled at random：Model group, psoralea corylifolia (low, high dose) group, vitamin E group, every group of 8 mouse, use D12451 height instead Fat forage feed.After 3 days, normal and model group gives 0.02ml/g physiological saline gavages；Psoralea corylifolia (low dosage) group is given 1.125mg/g psoralea corylifolia granule solution gavages；It is molten that psoralea corylifolia (high dose) group gives 2.25mg/g psoralea corylifolia granules Liquid gavage；Vitamin E group gives 0.01mg/g vitamin E soft capsule suspension oral gavages.Each group gastric infusion is to 10 week old.Put to death Preceding fasting 12 hours, eyeball method collecting blood sample is extractd, 30min is centrifuged under the conditions of blood sampling volume 2-3ml, 3000r/min, is separated Serum, -80 DEG C of preservations；Take liver organization：Left lateral lobe of liver, left middle lobe load cryopreservation tube, and -80 DEG C of preservations are standby；Liver right middle lobe, Right upper lobe and right lower lobe give 4% paraformaldehyde to fix, FFPE, are detected for hematoxylin-eosin (HE) and SABC.
Influence of the psoralea corylifolia of embodiment 1 to young age fatty liver mouse OGTT and HOMA-IR
1. the method for oral glucose tolerance test (OGTT)
OGTT experiments were carried out to the mouse of 3 days before execution first.Mouse fasting 12 hours, cut tail blood taking method collection blood mark This, detects fasting blood-glucose (FBG) value with Roche blood glucose meter/remarkable type, gives 50% glucose 2g/kg gavages afterwards, then again Cut tail and take blood, blood glucose meter determines 1h, 2h blood glucose (PG-1h, PG-2h) value after gavage respectively.
2. calculate the method for insulin resistance index (HOMA-IR)
1) determination of glucose oxidase fasting blood-glucose (FBG), ELISA method measure FPI (FINS).According to Formula：FBG (mmol/L) × FINS (mIU/L)/22.5 calculates HOMA-IR.
3. result
As illustrated in chart 1：Compared with normal group, model mice serum FBG, PG-2h, HOMA-IR have rise (p< 0.01)；Compared with model group, each dosage group of psoralea corylifolia, the vitamin E horizontal no difference of science of statistics of group serum PG -2h (p ＞ 0.05), but FBG, HOMA-IR have reduction (p in various degree<0.01, p<0.05), wherein psoralea corylifolia high dose group the biggest drop (p <0.01), the statistically significant (p of HOMA-IR relatively low-doses group<0.01).Each group mice serum PG-1h levels have no obvious poor Different (p ＞ 0.05).
As from the foregoing：Psoralea corylifolia can reduce model mice fasting blood glucose level, improve model mice insulin resistance.
Influence (mean ± SD) of the psoralea corylifolia of table 1 to young age fatty liver mouse OGTT
Compared with normal group▲▲p<0.01；Compared with model group△△p<0.01,△p<0.05；With psoralea corylifolia low dose group ratio Compared with * * p<0.01.
Influence of the psoralea corylifolia of embodiment 2 to young age fatty liver lipid of mice
1. method
Fasting 12 hours before mouse is put to death, then extracts eyeball method collecting blood sample, blood sampling volume 2-3ml, 3000r/min Under the conditions of centrifuge 30min, separate serum, enzymatic assays blood TC values, TG values, HDL-C values.
As shown in table 2：Compared with normal group, there is metabolism disorder of blood lipid, serum TG value, TC values and LDL-C in model mice Value significantly rise (p<0.01), and Serum HDL-C level substantially reduces (p<0.01)；Compared with model group, each dose of psoralea corylifolia Each blood fat individual event level of amount group, vitamin E group is improved in varying degrees (p<0.01, p<0.05), wherein psoralea corylifolia high dose Most obvious (the p of group improvement amplitude<0.01), the statistically significant (p of difference relatively low-dose group<0.01, p<0.05).
As from the foregoing：Psoralea corylifolia can reduce model mice blood TG, TC, LDL-C level, elevation model mouse blood HDL-C water It is flat, adjust model mice blood fat disorder.
Influence (mean ± SD) of the psoralea corylifolia of table 2 to young age fatty liver lipid of mice
Compared with normal group▲▲p<0.01；Compared with model group△△p<0.01,△p<0.05；With psoralea corylifolia low dose group ratio Compared with * * p<0.01, * p<0.05.
The psoralea corylifolia of embodiment 3 is on the morphologic influence of young age fatty liver mouse liver
1. method
A. material process：
Fasting 12 hours, takes liver organization, liver right middle lobe, right upper lobe and right lower lobe give 4% paraformaldehyde before mouse is put to death It is fixed, FFPE, detected for hematoxylin-eosin (HE) standby；
B. hematoxylin-eosin (HE) dyes
Take paraffin section, dimethylbenzene dewaxing, graded ethanol aquation, deionized water rinsing, brazilwood extract dyeing 1-5min, go from Sub- water rinses 2min, 1% hydrochloride alcohol differentiation 15sec, deionized water rinsing 1min, 1% ammoniacal liquor blueing 30sec, deionized water 1min, eosin stains 30sec, deionized water rinsing 30sec are rinsed, graded ethanol dehydration, dimethylbenzene is transparent, neutral tree after drying Glue mounting, in observation by light microscope.
2. result
As shown in Fig. 1 (A-E)：Fig. 1 normal group mouse lobuli hepatis structures are visible, and hepatic cell cords and hepatic sinusoid arrangement are normal. Compared with normal group, the obvious oedema of model group liver cell, denaturation, the obvious steatosis of liver cell, portal area fibr tissue and small Bile duct proliferation, more inflammatory cell infiltration.Compared with model group, each treatment group's pathology of livers has improvement, psoralea corylifolia height Dosage group lesion is most light.Concrete condition is under mirror：Psoralea corylifolia low dose group liver cell oedema, see in it and be dispersed in fat drips vacuole, converge Area under control fibr tissue is slight hyperplasia, inflammatory cell infiltration；Psoralea corylifolia high dose group part of hepatocytes mild fatty is denatured, portal area A little inflammatory cell infiltration；Vitamin E group portal area inflammatory cell infiltration, see in liver cell and be dispersed in fat drips vacuole.
As from the foregoing：Psoralea corylifolia can mitigate model mice hepatocellular injury, steatosis, improve portal area fibre modification and Inflammation.
Influence of the psoralea corylifolia of embodiment 4 to young age fatty liver mouse liver function and liver TG contents
1. method
A. material processing early stage：
Fasting 12 hours before mouse is put to death, extracts eyeball method collecting blood sample, blood sampling volume 2-3ml, 3000r/min condition Lower centrifugation 30min, separate serum, -80 DEG C of preservations；Take liver organization：Left lateral lobe of liver, left middle lobe load cryopreservation tube, -80 DEG C of guarantors Deposit.
B. enzymatic assays blood ALT, AST and hepatic tissue TG contents
2. result
As shown in table 3：Compared with normal group, model mice Serum ALT, AST have rise (p<0.01), liver TG contents Raise (p<0.01)；Compared with model group, each dosage group of psoralea corylifolia, vitamin E group Serum ALT, AST reduce (p<0.01), Liver TG contents reduce (p<0.01), the wherein obvious (p of the psoralea corylifolia high dose group relatively low-dose range of decrease<0.01, p<0.05).It is shown in Table 3。
As from the foregoing：Psoralea corylifolia can mitigate model mice liver function damage and reduce liver TG contents.
Influence (mean ± SD) of the psoralea corylifolia of table 3 to young age fatty liver mouse liver function and hepatic fat content
Compared with normal group▲▲p<0.01；Compared with model group△△p<0.01；The * * p compared with psoralea corylifolia low dose group< 0.01, * p<0.05
Influence of the psoralea corylifolia of embodiment 5 to young age fatty liver murine liver tissue CD44 protein expressions
1. method
A. material processing early stage：
Fasting 12 hours, takes liver organization before mouse is put to death：Liver right middle lobe, right upper lobe and right lower lobe give 4% paraformaldehyde It is fixed, FFPE, detected for SABC standby；
2. ImmunohistochemistryMethods Methods determine hepatic tissue CD44 protein expressions
Paraffin section is placed in baking piece 2h in 65 DEG C of baking ovens, dewaxing, aquation.After being washed with PBS, section is placed in EDTA and delayed Microwave method in fliud flushing, washed with PBS after natural cooling.Section is placed in 3% hydrogenperoxide steam generator again, lucifuge is incubated at room temperature Educate 10min.5% bovine serum albumin closing 20min after PBS washings dry.Confining liquid is removed, every section adds about 50 μ l dilutions (ratio 1:150) rabbit anti-mouse CD44 monoclonal antibodies covering tissue, 4 DEG C overnight.After being washed with PBS, every at 37 DEG C Section is incubated secondary antibody 50min.After PBS washings, every section plus the colour developing of Fresh DAB solution.Colour developing completely after, distilled water or Running water is rinsed, and haematoxylin is redyed, and 1% hydrochloride alcohol differentiation, running water rinses, and ammoniacal liquor returns indigo plant, and flowing water rinses.Section is by ladder Spend dehydration of alcohol to dry, dimethylbenzene is transparent, neutral gum sealing.
As shown in Fig. 2 (A-E)：Normal group murine liver tissue portal area lymphocyte, which is negative, expresses CD44.With normal group Compare, model group mouse portal area lymphocyte is in strong positive expression CD44.Compared with model group, each treatment group CD44 expression is equal It is reduced：Psoralea corylifolia low dosage mouse portal area part Expressions In Lymphocytes CD44；Psoralea corylifolia high dose group and vitamin E group are small Mouse portal area part lymphocyte weak expression CD44.
As from the foregoing：Psoralea corylifolia can lower model mice liver CD44 protein expressions, improve young age fatty liver mouse early stage Liver fibrosis.
Influence of the psoralea corylifolia of embodiment 6 to young age fatty liver murine liver tissue inflammatory factor
1. method
A. material processing early stage：
Fasting 12 hours, takes liver organization before mouse is put to death：Left lateral lobe of liver, left middle lobe load cryopreservation tube, -80 DEG C of preservations；
B.ELISA methods measure hepatic tissue TNF-α, IL-8 are horizontal.
2. result
As shown in table 4：Compared with normal group, model mice hepatic tissue TNF-α, IL-8 have rise (p<0.01)；With mould Type group compares, and each dosage group of psoralea corylifolia, vitamin E group hepatic tissue TNF-α, IL-8 reduce (p<0.01), wherein psoralea corylifolia is high Obvious (the p of the dosage group relatively low-dose range of decrease<0.01).
As from the foregoing：Psoralea corylifolia can reduce inflammation factor content, improve inflammation reaction.
Influence (mean ± SD) of the psoralea corylifolia of table 4 to young age fatty liver murine liver tissue inflammatory factor
Compared with normal group▲▲p<0.01；Compared with model group△△p<0.01；The * * p compared with psoralea corylifolia low dose group< 0.01
Active influence of the psoralea corylifolia of embodiment 7 to young age fatty liver murine liver tissue NF- κ B
1. method
A. material processing early stage：
Fasting 12 hours, takes liver organization before mouse is put to death：Left lateral lobe of liver, left middle lobe load cryopreservation tube, -80 DEG C of preservations；
B.Western blot methods measure hepatic tissue NF- κ B p65, phosphorylation NF- κ B p65 protein expressions
Hepatic tissue total protein is extracted, protein concentration, -80 DEG C of preservations are determined with BCA methods.40ug albumen is taken, is used after denaturation 10%SDS-PAGE separates gel electrophoresis, pvdf membrane transferring film, 5% skimmed milk power (non-phosphorylating albumen) or 0.5% bovine serum albumin (phosphorylated protein) is closed, and (GAPDH dilution ratios are 1 to 4 DEG C of incubation primary antibodies:10000；NF- κ B p65 dilution ratios are 1: 2000；Phosphorylation NF- κ B p65 dilution ratios are 1:1000) incubator overnight, after TBST washes film, secondary antibody 30min is incubated, TBST is washed After film, chemoluminescence method detection pvdf membrane, carrying out analysis gray value with AlphaEaseFC softwares, (p-p65/p65 ratios represent NF- κ B activity).
2. result
As shown in Figure 3：Compared with normal group, model mice hepatic tissue p-p65/p65 ratios significantly raise (p<0.01)；With Model group compares, and each dosage group of psoralea corylifolia, vitamin E group hepatic tissue p-p65/p65 ratios substantially reduce (p<0.01), wherein Obvious (the p of the psoralea corylifolia high dose group relatively low-dose range of decrease<0.05).
As from the foregoing：Psoralea corylifolia can lower hepatic tissue phosphorylation and non-phosphorylating NF- κ B p65 protein expression ratios, suppress NF-Kb is activated.
The preparation of the oral formulations of embodiment 8
Psoralea corylifolia, vitamin C and sucrose are taken by weight 1:3:1 ratio, is made oral formulations according to a conventional method.
The preparation of the tablet of embodiment 9
Psoralea corylifolia, fenugreek, sucrose and dextrin are taken by weight 1:1:3:1 ratio, is made tablet according to a conventional method.
The preparation of the granule of embodiment 10
Psoralea corylifolia, fenugreek, sucrose and dextrin are taken by weight 1:1:3:1 ratio, is made granule according to a conventional method.
Other unspecified parts are prior art.Although above-described embodiment is made that to the present invention and retouched in detail State, but it is only part of the embodiment of the present invention, rather than whole embodiments, people can also according to the present embodiment without Other embodiment is obtained under the premise of creativeness, these embodiments belong to the scope of the present invention.

Claims (10)

1. a kind of psoralea corylifolia is applied in the medicine for preparing treatment children's fatty liver.
2. application according to claim 1, it is characterised in that：Children's fatty liver is children's non-alcoholic fatty liver Scorching NASH.
A kind of 3. pharmaceutical preparation for treating children's nonalcoholic fatty liver disease NASH, it is characterised in that：Benefit containing effective dose Bone fat and pharmaceutically acceptable auxiliary material and/or auxiliary material.
4. the pharmaceutical preparation of children's nonalcoholic fatty liver disease is treated according to claim 3, it is characterised in that：The medicine Thing preparation is containing also fenugreek.
5. according to the pharmaceutical preparation of the treatment children's nonalcoholic fatty liver disease of claim 3 or 4, it is characterised in that：Institute Auxiliary material is stated as in vitamin C, sorbierite, mannitol, xylitol, fructose, amino acid, meglumine, dextrin, magnesium stearate and sucrose Any one or two kinds.
6. the pharmaceutical preparation according to claim 3 or 4, it is characterised in that：The pharmaceutical preparation is bowel lavage or oral formulations.
7. pharmaceutical preparation according to claim 5, it is characterised in that：The oral formulations are tablet, oral liquid or particle Agent.
8. pharmaceutical preparation according to claim 6, it is characterised in that：The oral formulations by 1 parts by weight psoralea corylifolia, 3 The vitamin C of parts by weight and 1 parts by weight of sucrose.
9. pharmaceutical preparation according to claim 6, it is characterised in that：Psoralea corylifolia of the tablet by 1 parts by weight, 1 weight The fenugreek, the sucrose of 3 parts by weight and the dextrin of 1 parts by weight of part.
10. pharmaceutical preparation according to claim 6, it is characterised in that：Psoralea corylifolia, 1 weight of the granule by 1 parts by weight Measure the fenugreek, the sucrose of 3 parts by weight and the dextrin of 1 parts by weight of part.
CN201710419208.0A 2017-06-06 2017-06-06 Psoralea corylifolia is applied in the medicine for preparing treatment children's fatty liver Pending CN107349252A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021074357A1 (en) * 2019-10-17 2021-04-22 Enyo Pharma Thiophene derivatives for use in treating portal inflammation and fibrosis

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1566114A (en) * 2003-06-16 2005-01-19 高和英 Novel psoralen derivative for annihilating live virus or germ

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1566114A (en) * 2003-06-16 2005-01-19 高和英 Novel psoralen derivative for annihilating live virus or germ

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
SEO EUNHUI，等: "Psoralea corylifolia L. Seed Extract Attenuates Nonalcoholic Fatty Liver Disease in High-Fat Diet-Induced Obese Mice", 《NUTRIENTS》 *
中华人民共和国卫生部药典委员会: "《中华人民共和国卫生部药品标准 中药成方制剂（第17册）》", 31 December 1998 *
兰卫，等: "葫芦巴药理作用的研究进展", 《西北药学杂志》 *
徐以亮: "补骨脂配方颗粒质量标准研究", 《药物鉴定》 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2021074357A1 (en) * 2019-10-17 2021-04-22 Enyo Pharma Thiophene derivatives for use in treating portal inflammation and fibrosis
CN114555577A (en) * 2019-10-17 2022-05-27 埃尼奥制药公司 Thiophene derivatives for the treatment of portal vein inflammation and fibrosis

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