CN104971090A - Application of cyclocarya paliurus effective part in preparation of medicine used for treating non-alcoholic fatty liver disease - Google Patents
Application of cyclocarya paliurus effective part in preparation of medicine used for treating non-alcoholic fatty liver disease Download PDFInfo
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Abstract
The invention belongs to the field of pharmaceuticals and relates to application of a cyclocarya paliurus effective part in preparation of health products and medicines which are used for treating a non-alcoholic fatty liver disease. The cyclocarya paliurus effective part mainly contains natural triterpenoid and can be used for effectively reducing rat liver lipid accumulation degree and triglyceride level which are caused by high fat diet, improving the liver histology and liver functions and improving the liver oxidative stress and the organism inflammatory level, so that the cyclocarya paliurus effective part can be used for obviously alleviating non-alcoholic fatty liver disease and preparing health products and medicines which are used for treating the non-alcohol fatty liver disease. The application of the cyclocarya paliurus effective part has the advantage that application range of cyclocarya paliurus preparation in the medical field.
Description
Technical field
The present invention relates to the novelty teabag of Cyclocarya paliurus Iljinskaja extract and effective site thereof, particularly relate to the novelty teabag of Cyclocarya paliurus Iljinskaja effective site at pharmaceutical field, is exactly the purposes in preparation control non-alcoholic fatty liver disease medicine.
Background technology
Non-alcoholic fatty liver disease (non-alcoholic fatty liver disease, NAFLD) a kind ofly stores up clinical syndrome for feature without excessive drinking history with hepatic parenchymal cells steatosis and fat.It is current modal chronic hepatopathy.Its basic course of disease comprises simple fatty liver, fat hepatitis, fatty liver fibrosis, finally may develop into fatty cirrhosis and fatty hepatocarcinoma, may cause too early death.About there is the NAFLD patient of 20 ~ 40% in the whole world, and quantity is constantly rising and presenting the trend of becoming younger.NAFLD is the reaction of liver to metabolism syndrome, closely bound up with the morbidity of obesity, type-II diabetes, cardiovascular disease, and obesity patient NAFLD prevalence is 65 ~ 95%, and type 2 diabetes mellitus patient NAFLD prevalence is 28 ~ 55% and 27 ~ 92%.Because NAFLD morbidity has invisible, unresolved, universality, and the hazardness of its pathological changes is comparatively large, therefore becomes the direction of world professional person effort to its sick control.
Change lifestyles, intervening diet and motion can effective control NAFLD, by the improvement of primary stage generally recommended in order to disease, but implements difficulty, often cannot get a desired effect, and to hepatic fibrosis, liver cirrhosis patient curative effect is not obvious.Therefore, effective Drug therapy is essential.Clinical in increasing insulin sensitivity, improving insulin resistant (as class biguanide, thiazolidinediones), reduce blood fat (Statins, the special class of shellfish), antioxidation (vitamin E) treats non-alcoholic fatty liver disease, but because of these medicines or there is side effect, or aggravation burden of liver, or drug effect is not good enough, uses and be restricted.Not yet there is the treatment NAFLD medicine of approval at present.Therefore, develop NAFLD medicine and health promoting product safely and effectively and there is important innovative significance and Clinical practice value.
Cyclocarya paliurus Iljinskaja (Cyclocarya paliurus Batal.) is Juglandaceae (Juglandaceae) cyclocarya plant, is distributed widely in the ground such as Jiangxi, Guangxi.It is a kind of tall and big fast-growing deciduous tree, and leaf sweetness is moistened, can promoting the production of body fluid to quench thirst, clearing away summer-heat.Leaf of Cyclocarya paliurus Iljinskaja among the people makes millet paste for a long time, and local people are long-lived, and rarely have metabolism class disease, be referred to as " Folium hydrangeae strigosae ", " refreshing tea ", visible Cyclocarya paliurus Iljinskaja has the multiple physiologically active useful to human body and pharmacological function, and has no side effect.Take Cyclocarya paliurus Iljinskaja as the health tea of raw material production be the health tea that the 1st, China obtains U.S. FDA accreditation.In recent years, Chinese scholars to the physical and chemical composition of Cyclocarya paliurus Iljinskaja and health care in animal level and clinical trial preliminary study, show that Cyclocarya paliurus Iljinskaja and relevant healthcare product have pharmacological action and the health-care effect of multiple beneficial, comprise blood pressure lowering, blood sugar lowering, fat-reducing, antitumor, antiallergic, enhancing human body immunity, the effect such as antibacterial.Research finds, Cyclocarya paliurus Iljinskaja can significantly improve body lipid metabolism, and play and reduce body weight, improve blood fat and reduce the effect that liver fat drips, and have good antiinflammatory antioxidant activity, prompting Cyclocarya paliurus Iljinskaja may play positive preventive and therapeutic effect to the generation development of NAFLD.
Summary of the invention
The present invention is directed to the current treatment status of NAFLD; a kind of novelty teabag of Cyclocarya paliurus Iljinskaja extract effective site is proposed; namely the application in preparation control non-alcoholic fatty liver disease medicine; Cyclocarya paliurus Iljinskaja effective site effectively can improve the NAFLD Hepaticlipid level that high lipid food causes; play a protective role to liver, the control for non-alcoholic fatty liver disease provides to be selected safely and efficiently.
Effective site involved in the present invention is extracted and is obtained from plant Cyclocarya paliurus Iljinskaja, and in this total effective parts, the content of total triterpene is 5 ~ 100%, and can prepare according to the method described in various document, these are all technology well-known to those skilled in the art.In addition, Cyclocarya paliurus Iljinskaja effective site is also bought by various commercial sources and is obtained, main containing following one or more:
Wherein: Ra: β-D-Xyl-(1 → 3)-α-L-Rha-(1 → 2)-α-L-Ara (p) Rb: α-L-Rha-(1 → 4)-β-D-Glu-(1 → 6)-β-D-Glu
Its preparation method is preferably, and comprises the following steps:
(1) extract: the dry medical material of leaf of Cyclocarya paliurus Iljinskaja does solvent extraction, merge extractive liquid, through 0%-100% ethanol or methanol (C1-C5 fatty alcohol), filters, concentrated.
Said extracted method can be: reflux, dipping, percolation, the multiple method such as ultrasonic.
(2) purification: this concentrated solution is by the means purification such as organic solvent (petroleum ether, ethyl acetate, chloroform, n-butyl alcohol etc.) extraction, Sephadex LH-20, macroporous resin, silica gel.
The present invention also provides its effective site of Cyclocarya paliurus Iljinskaja and pharmaceutically acceptable adjuvant or complementary composition to be prepared into pharmaceutically conventional preparation.
Provided by the invention with Cyclocarya paliurus Iljinskaja extract or its effective site for the obtained new drug of raw material, not only comprise single preparations of ephedrine, be also included in compound preparation and add Cyclocarya paliurus Iljinskaja effective site as the health product of active component and medicine.
Accompanying drawing illustrates:
Fig. 1. Cyclocarya paliurus Iljinskaja effective site HPLC-UV detection
Fig. 2. Cyclocarya paliurus Iljinskaja effective site is on the impact of the relative lipid level of High fat diet rats liver
Fig. 3. Cyclocarya paliurus Iljinskaja effective site feeds the impact of Hepaticlipid accumulation to high fat
Embodiment 1: the preparation of Cyclocarya paliurus Iljinskaja effective site
1. the preparation of Cyclocarya paliurus Iljinskaja effective site
Cyclocarya paliurus Iljinskaja dried leaves 1kg, through 80% (volume/volume) ethanol 8L reflux, extract, 3 times, each 2h, merge extractive liquid, filter, recycling design obtains extractum.After extractum adds suitable quantity of water suspendible, with 1.5 times amount chloroform extraction 3 times, recycling design obtains Cyclocarya paliurus Iljinskaja extract.Wherein the percentage composition of triterpene is 10%-80% (w/w).
2. the mensuration of total triterpene contents:
The preparation of standard substance: precision takes oleanolic acid standard substance 5mg, prepares concentration is the mother solution of 0.1mg/mL with dehydrated alcohol.Respectively get mother solution 1.0 ~ 5.0mL again, be settled to 10mL with dehydrated alcohol respectively.Draw each 1mL of reference substance solution after above-mentioned dilution, volatilize solvent, add 5% vanillin-glacial acetic acid 0.4mL and perchloric acid 0.8mL, mix homogeneously, cool rapidly heat 10min in 80 DEG C of water-baths after, finally add the glacial acetic acid of 5mL, survey absorbance in 550nm place.Prepared by blank control sample: only add successively and the vanillin-glacial acetic acid of standard substance same concentrations, perchloric acid and glacial acetic acid each 0.4,0.8 and 5mL.With concentration to absorbance drawing standard curve, and calculate standard regressive method.
The mensuration of total triterpene contents in sample: the leaf of Cyclocarya paliurus Iljinskaja of accurately weighed 5g refines triterpenic acid powder 5mg in 50mL volumetric flask, and anhydrous alcohol solution is also diluted to graduation mark, obtains sample liquid.Pipette samples liquid 1mL, volatilizes solvent, by the sextuplicate mensuration absorbance (Abs) of method under standard curve item, calculates the content of total triterpene.
3.HPLC analyzes
Cyclocarya paliurus Iljinskaja effective site position sample is appropriate, is dissolved in AG methanol, obtains the sample solution of 1mg/mL concentration with filtering with microporous membrane.The chromatographic condition of this experiment is: adopt Alltima C18 column C18 chromatographic column (5 μm, 4.6mm × 250mm) to take acetonitrile as mobile phase A, water-0.02% formic acid is Mobile phase B; Adopt A-B mixed flow phase gradient eluting: 0 ~ 20min, 50%A; 20 ~ 22min, 50% ~ 53%A; 22 ~ 40min, 53%A; 40 ~ 55min, 53% ~ 100%A; 55 ~ 60min, 100%A.Flow velocity is 1mLmin-1, column temperature 27 DEG C, and determined wavelength is 205nm, sample size 10 μ L.By comparing its retention time determination chromatographic peak with standard substance, (1) arjunolic acid (arjunolic acid), (2) cyclocarya paliurus acid B (cyclocaric acid B), (3) pterocaryoside B, (4) 3 β, 23-dihydroxy-12-alkene-28-ursolic acid (3 β, 23-dihydroxy-12-ene-28-ursolic acid), (5) helexin (hederagenin), (6) oleanolic acid (oleanolic acid) (as Fig. 1).
Embodiment 2: Cyclocarya paliurus Iljinskaja effective site is to the improvement result of NAFLD
1. material
1.1 animal
Healthy male SD rat, 180 ~ 220g, SPF level, purchased from Nantong University's Experimental Animal Center, credit number: SCXK (SU12014-000).
1.2 medicines and reagent
Cyclocarya paliurus Iljinskaja effective site powder, is dissolved in appropriate 0.5% carboxymethylcellulose sodium solution, prepares certain density solution for standby.Simvastatin is purchased from Changsha, Yichang City pharmaceutcal corporation, Ltd.Triglyceride (TG), T-CHOL (TC), non-esterified fatty acid (NEFAs), glutamate pyruvate transaminase (ALT), glutamic oxaloacetic transaminase, GOT (AST), alkali phosphatase (ALP), malonaldehyde (MDA) detection kit build up Bioengineering Research Institute purchased from Nanjing.Rat TNF-α Elisa test kit is purchased from Nanjing Sen Beijia Bioisystech Co., Ltd.
2. experimental technique
After rat adaptability is fed, be divided into two groups at random, blank group (NC group) 10, gives normal diet and light water, remaining modeling group 30, gives high lipid food (cholesterol 2%, Adeps Sus domestica 10%, egg yolk 10%, sodium cholate 0.5%, normal feedstuff 77.5%).Modeling is after six weeks, and high fat is fed rat and is divided into 3 groups at random, hyperlipidemia model group (HC group), Cyclocarya paliurus Iljinskaja effective site administration group (ChE group), simvastatin positive drug group (SMT group).
Within 7th week, rise, each group rat continues to feed former feedstuff 4 weeks, and every day gavage sodium carboxymethyl cellulose suspension.NC group and HC group gavage every day 0.5% carboxymethylcellulose sodium solution.ChE group gavage every day Cyclocarya paliurus Iljinskaja ethanol extract (dosage make a living dose 8g/kg), SMT group gavage every day simvastatin (4mg/kg).Often group get three rats weekly by Magnetic Resonance Spectrum (
1h-MRS) liver is detected relative to lipid content (liver relative lipid content=fat peak/(fat peak+water peak), wherein the chemical shift at water peak is 4.7ppm, and the chemical shift at fat peak is 2.1,1.2,0.9ppm).
After experiment terminates, all Rat Fast 12 hours, weigh, and ventral aorta is taken a blood sample, separation of serum, detect TC, TG, NEFA in serum ' s, ALT, AST, ALP, TNF-α.Collect whole liver, weigh and calculate liver coefficient, liver TC, TG, NEFA ' s, TNF-α, MDA, and carry out hepatic pathology sections observation.(H & E dyes).
3. result
The change that 3.1 rat body weight regulating liver-QI are heavy
As shown in table 1, compared with NC group, the body weight of HC group rat significantly raises (p < 0.01).Compared with HC group rat, ChE group and SMT group rat body weight all significantly reduce (p < 0.05).Compared with NC group, HC group rat liver coefficient significantly increases (p < 0.05), and ChE, SMT administration significantly reduces the liver coefficient (p < 0.05) that high fat feeds rat.Illustrate that Cyclocarya paliurus Iljinskaja effective site significantly can reduce high fat and feed rat body weight, alleviate liver enlargement.
The impact that table 1. Cyclocarya paliurus Iljinskaja effective site weighs high fat nursing rat body weight and liver
The change of 3.2 rat fat content
As shown in table 2, compare with NC group, high fat feeds Serum TC, and TG level significantly increases (p < 0.01).After ChE and SMT administration, rat blood serum TG level all significantly declines (p < 0.01).ChE administration significantly reduces Serum TC level (p < 0.05), and SMT group Serum TC level reduces significantly (p < 0.01).High fat diet rats serum N EFAs level comparatively NC group raises significantly.After ChE, SMT administration, Induced by High Fat Diet in Rats serum N EFAs all significantly reduces (p < 0.05).The above results illustrates that Cyclocarya paliurus Iljinskaja administration can effectively prevent high fat to feed the blood fat disorder caused.
Table 2. Cyclocarya paliurus Iljinskaja effective site feeds the impact of Serum Lipids in Experimental HypercholesterolemicRats to high fat
3.3 Liver Function index changes
As shown in table 3, HC group rat ALT, AST, ALP level comparatively NC group significantly raises (p < 0.01), and HC group is compared in Cyclocarya paliurus Iljinskaja administration, ALT, AST, ALP significantly reduce (p < 0.01), and in dose dependent.Illustrate and the hepatic injury that Cyclocarya paliurus Iljinskaja administration effectively can improve high fat diet and causes improve liver function.
Table 3. Cyclocarya paliurus Iljinskaja effective site feeds the impact of Liver Function to high fat
The change of 3.4 Hepaticlipid levels
As shown in table 4, high lipid food is fed and is significantly improved rat liver TC, TG level (p < 0.01).After ChE and SMT administration, compare HC group, rat liver TC level significantly reduces (p < 0.05), and TG level extremely significantly reduces (p < 0.01).HC rat liver NEFAs level comparatively NC group raises significantly.After ChE, SMT administration, Induced by High Fat Diet in Rats liver NEFAs all significantly reduces (p < 0.05).The above results illustrates that Cyclocarya paliurus Iljinskaja administration effectively can reverse high fat and feed the liver lipids disorder caused.
Table 4. Cyclocarya paliurus Iljinskaja effective site is on the impact of Hepaticlipid level
The relative lipid content change of 3.5 liver
Fig. 2 shows 6 ~ 10 weeks HC groups and ChE stack features Hepaticlipid wave spectrogram.Can significantly see in figure, 6 weeks time, the apparition of rat liver fat, and ChE administration significantly reduces liver lipids level.All spectral datas are carried out quantitatively and adds up, found that, High fat diet feeds the rat of 6 weeks, there was no significant difference between each group, but compared with the rat fed with chow diet, the relative lipid content of liver significantly raises (p < 0.01).Pharmaceutical intervention has significantly reversed liver lipids content.SMT administration is after 1 week, liver relative to lipid content comparatively modeling terminate remarkable reduction (p < 0.01), and administration ChE is after 2 weeks, liver also has remarkable reduction (p < 0.01) before comparing administration relative to lipid content.After surrounding administration, ChE group lipid level there was no significant difference relative to NC group liver.
3.6 liver histological inspections
Fig. 3 is the rat liver H & E coloration result of feature.Compare normal liver tissue, HC group shows the hepatocyte swollen, irregular liver rope arrangement, and a large amount of fat drips.But ChE administration improves liver structure dramatically, decrease the liver lipids accumulation that high fat diet causes.
3.7 inflammatory parameters changes
As shown in table 5, compare NC group, HC group rat blood serum and liver TNF-alpha levels significantly improve (p < 0.05).And ChE group and SMT group rat blood serum and liver TNF-alpha levels are significantly lower than HC group rat (p < 0.05).
Table 5. Cyclocarya paliurus Iljinskaja effective site feeds the impact of rat TNF-alpha levels to high fat
3.8 hepatic lipid peroxidation thing changes of contents
As shown in table 6, compare NC group, HC group rat blood serum and liver MDA level significantly improve (p < 0.05).And ChE group and SMT group rat blood serum and liver MDA level are significantly lower than HC group rat (p < 0.05).
Table 6. Cyclocarya paliurus Iljinskaja effective site feeds the impact of Peroxidation In Rat Liver thing content to high fat
Claims (6)
1. the application of Cyclocarya paliurus Iljinskaja effective site in preparation control non-alcoholic fatty liver disease medicine.
2. the effective site of Cyclocarya paliurus Iljinskaja described in claim 1, is characterized in that: this Cyclocarya paliurus Iljinskaja effective site is extracted and obtained from Cyclocarya paliurus Iljinskaja, and the triterpenoid content of this effective site is 5 ~ 100%.
3. the effective site described in claim 1, is characterized by, and obtains in Cyclocarya paliurus Iljinskaja plant, and mainly contain following triterpenoid one or more, its structure is:
Wherein: Ra: β-D-Xyl-(1 → 3)-α-L-Rha-(1 → 2)-α-L-Ara (p) Rb: α-L-Rha-(1 → 4)-β-D-Glu-(1 → 6)-β-D-Glu
4. the effective site of Cyclocarya paliurus Iljinskaja described in right 1-4, its preparation method is:
(1) extract: the dry medical material of leaf of Cyclocarya paliurus Iljinskaja does solvent extraction, merge extractive liquid, through 5%-100% ethanol or methanol (C1-C5 fatty alcohol), filters, recycling design, concentrated.
Said extracted method can be: the multiple methods such as reflux, dipping, percolation, supersound extraction.
(2) purification: by the means purification such as organic solvent (petroleum ether, ethyl acetate, chloroform, n-butyl alcohol etc.) extraction, Sephadex LH-20, macroporous resin, silica gel.
5. prevent and treat non-alcoholic fatty liver disease medicine described in claim 1, it is characterized in that, described medicine also comprises Cyclocarya paliurus Iljinskaja effective site and pharmaceutically acceptable adjuvant or complementary composition and is prepared into pharmaceutically conventional preparation.
6. prevent and treat non-alcoholic fatty liver disease medicine described in claim 1 and 6, it is characterized in that, not only comprise with the obtained single preparations of ephedrine of Cyclocarya paliurus Iljinskaja effective site, be also included in compound preparation and add Cyclocarya paliurus Iljinskaja effective site as the health product of active component and medicine.
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| CN106511353A (en) * | 2016-09-08 | 2017-03-22 | 中国药科大学 | Preparation and application of triterpene drug for inhibiting synthesis and secretion of apoB48 in intestinal tracts |
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| CN108186723A (en) * | 2018-03-15 | 2018-06-22 | 广西中医药大学 | Blue or green money willow extract and its application of preparation and reducing blood lipid |
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| CN111529569A (en) * | 2020-05-22 | 2020-08-14 | 广州医科大学附属第三医院（广州重症孕产妇救治中心、广州柔济医院） | Composition for treating non-alcoholic fatty liver disease and application |
| CN112190587A (en) * | 2020-10-26 | 2021-01-08 | 江苏省中医药研究院 | Cyclocarya paliurus saponin and derivative thereof and pharmaceutical application of salt or precursor compound thereof |
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