CN104000862A - Application of total flavonoids of herba epimedii in preparing medicine for preventing hepatic fibrosis - Google Patents
Application of total flavonoids of herba epimedii in preparing medicine for preventing hepatic fibrosis Download PDFInfo
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- CN104000862A CN104000862A CN201410258438.XA CN201410258438A CN104000862A CN 104000862 A CN104000862 A CN 104000862A CN 201410258438 A CN201410258438 A CN 201410258438A CN 104000862 A CN104000862 A CN 104000862A
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- herba epimedii
- total flavones
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Abstract
The invention relates to application of total flavonoids of herba epimedii in medicine preparing, in particular to application of the total flavonoids of herba epimedii in preparing the medicine for preventing hepatic fibrosis. The medicine is a common preparation in clinic and made of the total flavonoids of herba epimedii and a convention amount of auxiliary materials accepted medically. The medicine has the inhibiting effect on hepatic fibrosis, and can reduce the jaundice level and the liver injury degree of a hepatic fibrosis patient.
Description
Technical field
The present invention relates to medicinal preparation, be specifically related to contain the not medicine of definite structure from Berberidaceae barrenwort.
Background technology
Herba Epimedii is herbaceos perennial, there is medical value widely, have another name called Herba Epimedii, its medicinal history is long, and Li Shizhen (1518-1593 A.D.) claims its useful vital essence, hard muscles and bones, mends the effect of waist knee joint, heart tonifying power in < < Compendium of Materia Medica > >.The demonstration of modern chemistry composition Study, the main effective ingredient of Herba Epimedii is Herba Epimedii total flavones and epimedium brevicornum polysaccharide, modern pharmacology research shows that Herba Epimedii total flavones has Promote immunity function, participates in the functions such as bone metabolism, antitumor, enhancing cardiovascular activity.
Herba Epimedii total flavones is a kind of mixture, mainly various icariin, icariside, consists of.Due to Herba Epimedii total flavones complicated component, so its new therapeutic use still awaits further research.
Hepatic fibrosis be comprise that most of hepatopathy of hepatitis, fatty liver, alcoholic liver etc. develops into liver cirrhosis, hepatocarcinoma must be through link.At present, the interferon that the drug main of western medical treatment hepatic fibrosis is wanted, but it is expensive, and side effect is large; Chinese medicine has low price, and the advantage that side effect is little, is more easily accepted by patient.Under the theoretical guidance that the traditional Chinese medical science is controlled in dialectical opinion, the research for the treatment of hepatic fibrosis obtains greater advance, as the patent medicine such as supporting vital QI and dispersing blood stasis 319 capsules, compound recipe 861 electuaries, FUFANG BIEJIA RUANGAN PIAN can obviously alleviate hepatic fibrosis, the reversion rate of hepatic fibrosis reaches 30%～71%.But, due to the complexity of liver cirrhosis pathology mechanism, still need to develop medicine evident in efficacy, toxic and side effects is little.
Summary of the invention
Technical problem to be solved by this invention is to provide the new purposes of Herba Epimedii total flavones, and Herba Epimedii total flavones is in the application of pharmacy.
Specifically, provide the application of Herba Epimedii total flavones at the medicine of preparation treatment hepatic fibrosis.
Application of the present invention, wherein said medicine is the common preparation clinically that the medically acceptable adjuvant of Herba Epimedii total flavones and conventional amount used is made, as, tablet, capsule or drop pill.
Herba Epimedii total flavones of the present invention can adopt the method for separating flavone from plant of seeing to obtain, and the method that the inventor recommends is as follows:
Get epimedium herb, add the ethanol extraction 3 times that 4～10 times of volumetric concentrations are 70%～90, filter, after merging filtrate, concentrating under reduced pressure becomes clear paste, add the ethanol that 4-10 times of volumetric concentration is 40%-60%, stir, stratification, filters, reclaim ethanol extremely without placing after alcohol taste, separate out precipitation, filter, obtain Herba Epimedii total flavones.
Medicine of the present invention is inhibited to hepatic fibrosis, can reduce jaundice level and the hepatic injury degree of patients with liver fibrosis.
The specific embodiment
Embodiment 1:
(1) extraction of Herba Epimedii total flavones is separated
Parallel 9 parts of the epimedium herbs of getting,, carry out reflux, extract, with 70%, 80%, 95% ethanol respectively, three parts of each concentration by 1 kilogram every part.Extract 2 times, each alcohol adding amount is 8 liters, filters, after filtrate merges, concentrating under reduced pressure becomes clear paste, then adds the ethanol of the 40%-60% that 4-10 doubly measures, stirs, stratification, filter, filtrate is recycled to without the water adding after alcohol taste with crude drug amount identical in quality, mix homogeneously, place, separate out precipitation, filter, obtain Herba Epimedii total flavones.
(2) mensuration of total flavones in epimedium flavone extract
1. instrument and reagent
1.1 instrument
Agilent 8483 ultraviolet-visible spectrophotometers (U.S. Agilent company); CP225 electronic balance (Sartorius AG)
1.2 reagent and sample
Icariin reference substance (lot number: 110737-200312 Nat'l Pharmaceutical & Biological Products Control Institute), methanol (analytical pure, Tianjin great Mao chemical reagent factory), distilled water (Watson)
2 methods
2.1 epimedium flavone extract extract sample treatment
The about 100mg of epimedium flavone extract that takes step (1) gained, is placed in 100ml measuring bottle, adds appropriate dissolve with methanol and is settled to scale, shakes up, and 0.45 μ m microporous filter membrane filters, and gets subsequent filtrate, to be determined.
The drafting of 2.2 ultraviolet spectrophotometry standard curves
2.2.1 the preparation of reference substance storing solution: precision takes the icariin 10.11mg that phosphorus pentoxide is dried to constant weight, is placed in 10ml measuring bottle, with dissolve with methanol and be settled to scale, shakes up, and obtains icariin reference substance storing solution.
2.2.2 the preparation of reference substance solution: the accurate 2.2.1 icariin reference substance storing solution 2.5ml that draws, be placed in 25ml measuring bottle, with dissolve with methanol and be settled to scale, shake up, obtaining concentration is 101.1 μ g/ml icariin reference substance solution.
2.2.3 the drafting of standard curve
Accurate reference substance solution 0.25,0.50,1.00,1.50,2.00, the 2.50ml of drawing, is placed in respectively 10ml measuring bottle, with methanol, is diluted to scale.Take methanol as blank, the trap with determined by ultraviolet spectrophotometry at 270nm place.The results are shown in Table 1.
Table 1 reference substance icariin and absorbance
With trap value, A returns icariin concentration C, obtains regression equation to be: y=0.4006x+0.1818
R2=0.9994, the range of linearity 2.53～25.28 μ g/ml.
2.3.4 precision test
The accurate reference substance solution 1ml that draws, totally 5 parts, be placed in respectively 10ml volumetric flask, according to method under Specification Curve of Increasing item, operate, retinue reagent blank, in 270nm wavelength, place measures its absorbance.The results are shown in Table 2.Result shows that this method has good precision.
Table 2 precision test result
2.3.5 repeatability is investigated
It is appropriate that precision takes Herba Epimedii total flavones extract, 6 parts, according to 2.1 lower contents, makes need testing solution.With 0.45 μ m microporous filter membrane, filter, get subsequent filtrate, measure.The results are shown in Table 3.Result shows that this method has breasting the tape property preferably.
Table 3 replica test result
2.3.6 average recovery test
Totally 6 parts of Herba Epimedii total flavones extract 0.5mg that get known content, are placed in respectively 25ml measuring bottle, and precision adds reference substance solution 1.0ml, by methanol constant volume, to scale, according to method under 2.2.3 item, measure, and the results are shown in Table 4.Result shows that this method has the good response rate.
Table 4 average recovery result of the test
2.3.7 sample determination
Precision takes epimedium flavone 2.0mg3 part prepared by Different Extraction Method, is placed in respectively 25ml measuring bottle, with methanol, is diluted to scale.At 270nm place, measure respectively trap, trap substitution standard curve is calculated to the content of total flavones in sample.Purity is calculated (lower same) with icariin.Concrete outcome is in Table 5.
Table 5 Herba Epimedii total flavones extracts experiment and result
Embodiment 2:
(1) preparation of epimedium flavone extract
Parallel 10 kilograms of the epimedium herbs of getting, carry out reflux, extract, three times with 70% ethanol, add 8 times of amounts of ethanol at every turn, filter, after filtrate merges, concentrating under reduced pressure becomes clear paste, then adds 45% ethanol of 8 times of amounts, stir, stratification, filters, filtrate is recycled to without the water adding after alcohol taste with crude drug amount identical in quality, mix homogeneously, places, and separates out precipitation, filter, obtain 63.5 grams of Herba Epimedii total flavones.
(2) mensuration of total flavones in epimedium flavone extract
The epimedium flavone extract that adopts the identical method of example 1 to obtain this routine described method is measured, and in described extract, general flavone content is, content is 63.1%
(3) preparation of icariin total flavones tablet
1. write out a prescription:
The epimedium flavone extract 500mg of step (1) gained, starch 200g.
2. preparation method
Extract powder is added to starch, mix, granulation, 60 ℃ are following dry, add moderate lubrication agent, are pressed into 1000 (0.2g/ sheets), and film coating, obtains.
Embodiment 3:
(1) preparation of epimedium flavone extract
Parallel 10 kilograms of the epimedium herbs of getting, carry out reflux, extract, three times with 80% ethanol, add 6 times of amounts of ethanol at every turn, filter, after filtrate merges, concentrating under reduced pressure becomes clear paste, then adds 50% ethanol of 6 times of amounts, stir, stratification, filters, filtrate is recycled to without the water adding after alcohol taste with crude drug amount identical in quality, mix homogeneously, places, and separates out precipitation, filter, obtain 62 grams of Herba Epimedii total flavones.
(2) mensuration of total flavones in epimedium flavone extract
The epimedium flavone extract that adopts the identical method of example 1 to obtain this routine described method is measured, and in described extract, general flavone content is, content is 61.3%
(3) preparation of epimedium flavone capsule
1. write out a prescription:
The epimedium flavone extract 500mg of step (1) gained, dextrin 250g.u
2. preparation method
Get epimedium flavone 500mg, add dextrin 250g, mix, pack capsule into No. 2, make 1000 of hard capsules (0.26g/ grain).
Embodiment 4: the effect of Herba Epimedii total flavones to common bile duct caused by ligature rat liver fibrosis
Getting Herba Epimedii total flavones in embodiment 3 tests.
50 of SD rats, SPF level, male and female half and half, are provided by Nanfang Medical Univ's Experimental Animal Center.Animal random packet, and give as follows medicine.
Normal group: distilled water.
Model control group: adopt common bile duct ligation to cause Rat Liver Fibrosis Model, give equal-volume distilled water.
Herba Epimedii total flavones low dose group: get Herba Epimedii total flavones before experiment, being mixed with concentration with 0.5%CMC-Na aqueous solution is 1.0mg/ml suspension.
Herba Epimedii total flavones high dose group: get Herba Epimedii total flavones before experiment, being mixed with concentration with 0.5%CMC-Na aqueous solution is 3.0mg/ml suspension.
3,5,7-trihydroxy-2-(4-methoxy-phenyl)-8-(3-methyl-but-2-enyl)-chromen-4-one dosage group: get 3,5,7-trihydroxy-2-(4-methoxy-phenyl)-8-(3-methyl-but-2-enyl)-chromen-4-one before experiment, being mixed with concentration with 0.5%CMC-Na aqueous solution is 3.0mg/ml suspension.
Modeling is respectively organized rat with 4ml/kg dosage, lumbar injection 10% chloral hydrate solution is anaesthetized, expose bile duct under liver, ligation liver and bile duct intersection and bile duct and pancreas intersection, wipe out to cause complete bile duct obstruction rat model by the bile duct in the middle of two places; The Normal group processing that do not operate.From modeling same day, Herba Epimedii total flavones is low, high dose group and 3,5,7-trihydroxy-2-(4-methoxy-phenyl)-8-(3-methyl-but-2-enyl)-chromen-4-one group are pressed 1ml/100g body weight gastric infusion 1 time, continuous 28 days every day.After last administration 1h, each is organized rat and slightly anaesthetizes with ether, and heart blood sampling is got serum and adopted test kit to detect TBIL, DBIL, ALT, AST, HA, LN, PC-III, TNF-alpha levels; Then open abdominal cavity and take out rapidly hepatic tissue and clean under normal saline, after homogenate, adopt test kit to detect SOD, MDA, GSH-Px, Hyp, adopt ELISA method to detect the expression of MMP-1 and TIMP-1 albumen.
(1) impact of Herba Epimedii total flavones on liver function index TBIL, DBIL, ALT, AST in serum
Herba Epimedii total flavones is low, level and the model control group comparison of TBIL, DBIL, ALT and AST in high dose group rat blood serum, and difference all has statistical significance (P<0.01 or 0.05), and is dose-effect relationship (in Table 2).Illustrate that Herba Epimedii total flavones can reduce jaundice level and the hepatic injury degree of common bile duct caused by ligature hepatic fibrosis rats.The jaundice level of 3,5,7-trihydroxy-2-(4-methoxy-phenyl)-8-(3-methyl-but-2-enyl)-chromen-4-one group reduction common bile duct caused by ligature hepatic fibrosis rats and hepatic injury degree are lower than the epimedium flavone group of same dose.
The impact of table 2 on liver function index TBIL, DBIL, ALT, AST in serum
Note: with Normal group comparison:
##p < 0.01; With model control group comparison:
*p < 0.05
*p < 0.01
(2) impact of Herba Epimedii total flavones on hepatic fibrosis index PC-III, HA, LN, Hyp
Herba Epimedii total flavones is low, Hyp content and model control group comparison in PC-III, HA, LN content and hepatic tissue in high dose group rat blood serum, difference all has statistical significance (P<0.01 or 0.05), and be dose-effect relationship (in Table 3), illustrate that Herba Epimedii total flavones is inhibited to common bile duct caused by ligature rat liver fibrosis.The impact of 3,5,7-trihydroxy-2-(4-methoxy-phenyl)-8-(3-methyl-but-2-enyl)-chromen-4-one group on hepatic fibrosis index PC-III, HA, LN, Hyp, suitable with epimedium flavone low dose group, illustrate that Herba Epimedii monomer suppresses the activity of hepatic fibrosis lower than the epimedium flavone of same dose.
The impact of table 3 on hepatic fibrosis index PC-III, HA, LN, Hyp
Note: with Normal group comparison:
##p < 0.01; With model control group comparison:
*p < 0.05
*p < 0.01
(3) impact of Herba Epimedii total flavones on hepatic tissue MMP-1 and TIMP-1 protein expression
Herba Epimedii total flavones is low, high dose group liver tissues of rats TIMP-1 protein expression and model control group comparison, and difference all has statistical significance (P<0.01 or 0.05), and is dose-effect relationship (in Table 4).Illustrate that Herba Epimedii total flavones can suppress the TIMP-1 protein expression of Liver Fibrosis Model rat liver, significantly promotes the decomposition of extracellular matrix in liver.And the ability of 3,5,7-trihydroxy-2-(4-methoxy-phenyl)-8-(3-methyl-but-2-enyl)-chromen-4-one group inhibition TIMP-1 protein expression is only suitable with low dose group.
The impact of table 4 on hepatic tissue MMP-1 and TIMP-1 protein expression
Note: with Normal group comparison:
##p < 0.01; With model control group comparison:
*p < 0.05
*p < 0.01
Claims (4)
1. Herba Epimedii total flavones is in the application of the medicine of preparation treatment hepatic fibrosis.
2. application according to claim 1, is characterized in that, described medicine is the common preparation clinically that the medically acceptable adjuvant of Herba Epimedii total flavones and conventional amount used is made.
3. application according to claim 2, is characterized in that, described medicine is tablet, capsule or drop pill.
4. a preparation method for the Herba Epimedii total flavones described in claim 2 or 3, the method is comprised of following steps: get epimedium herb, add the ethanol extraction 3 times that 4～10 times of volumetric concentrations are 70%～90, filter, after merging filtrate, concentrating under reduced pressure becomes clear paste, adds the ethanol that 4-10 times of volumetric concentration is 40%-60%, stirs, stratification, filter, reclaim ethanol to without placing after alcohol taste, separate out precipitation, filter, obtain Herba Epimedii total flavones.
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| CN112870211A (en) * | 2021-02-05 | 2021-06-01 | 遵义医科大学 | Application of icariside II in preparation of medicine for preventing and treating acute liver injury |
| CN113171385A (en) * | 2021-06-11 | 2021-07-27 | 劲牌持正堂药业有限公司 | Low-ash epimedium extract and preparation method thereof |
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| CN113171385A (en) * | 2021-06-11 | 2021-07-27 | 劲牌持正堂药业有限公司 | Low-ash epimedium extract and preparation method thereof |
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