CN101912435B - Leech extract and application thereof - Google Patents
Leech extract and application thereof Download PDFInfo
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- CN101912435B CN101912435B CN2010102495015A CN201010249501A CN101912435B CN 101912435 B CN101912435 B CN 101912435B CN 2010102495015 A CN2010102495015 A CN 2010102495015A CN 201010249501 A CN201010249501 A CN 201010249501A CN 101912435 B CN101912435 B CN 101912435B
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Abstract
The invention discloses a manyflower tickclover herbal extract and application thereof, relating to Chinese herbal medicinal extracts and application thereof. A preparation method of the manyflower tickclover herbal extract comprises the steps of: taking roots, stems or leaves of the Chinese herbal medicinal manyflower tickclover herbs as raw materials, carrying out leaching by refluxing on the raw materials with 75-95% (V/V) of ethanol, and recovering the solvent ethanol to obtain ethanol total extractum; and dispersing the total extractum with water, extracting with ethyl acetate, and recovering an extracting agent to obtain an ethyl acetate extract. Tests prove that the extract has better effect on preventing and treating fatty liver diseases caused by hyperlipidemia and hyperlipidemia syndrome and can be used for preparing medicaments for preventing and treating the fatty liver diseases caused by the hyperlipidemia and the hyperlipidemia syndrome; and compared with the traditional medicament, the medicament prepared from the invention has the advantages of low toxicity, safety, reliability, strong drug action, wide resources, simple process and low cost.
Description
Technical field: the present invention relates to Chinese herbal medicine extract and application thereof, specifically manyflower tickclover herbal extract and application thereof.
Background technology: Herba desmodii multifloi has another name called red palm grass, Radix Sophorae Tonkinensis, mashed cajaput, Radix Campylotropis Hirtella (Herba Myrsines Africanae), beggarweed, sticking body grass etc., is the Herb of pulse family beggar-ticks plant Herba desmodii multifloi Desmodium Multiflorum DC..Have promoting blood circulation and stopping pain, the effect of removing toxic substances and promoting subsidence of swelling cures mainly epigastric pain, infantile malnutrition, women's emaciation due to blood disorder, lumbar sprain, wound, urethritis, parotitis, lymphadenitis and venom etc.
Hyperlipemia is one of common disease of middle-aged and elderly people, also is to receive much attention and have a strong impact on the middle-aged and elderly people disease of orthobiosis.Because making one or more lipids in the blood plasma be higher than unusually, lipid metabolism or running normally be called hyperlipemia.Can show as hypercholesterolemia, hypertriglyceridemia, or both have concurrently.Blood fat is too high to be the risk factor that quickens in a plurality of factors of atherosclerosis.In addition, hyperlipemia also is an important risk factor that promotes hypertension, impaired glucose tolerance, diabetes.Hyperlipemia also can cause fatty liver, liver cirrhosis, cholelithiasis, pancreatitis, retinal hemorrhage, blind, peripheral vascular disease, limping, hyperuricemia etc.
Liver is the center organ of general metabolism and nutrition, and the synthetic and decomposition of carbohydrate, protein (aminoacid), fat is main to be accomplished through liver.Simultaneously, have multivitamin through liver synthesize, activation and storage.In addition, liver also has the biotransformation function, and most of endogenous and exogenous metabolism dead end product can excrete with the form of safety after need transforming through liver.Hyperlipidemia is the main cause that causes fatty liver: triglyceride raises and causes triglyceride acatharsia in the liver in the blood plasma, forms the liver inner lipid and stores up, and causes fatty liver.And hepatic disease is one of the most common disease that influences human health.At present, the treatment to fatty liver is still global severe problem.
Evidence; Manyflower tickclover herbal extract is having good effect aspect the control hepatic injury; Guizhou University has applied for patent of invention " a kind of manyflower tickclover herbal extract " application number 200910102986.2; This disclosure of the Invention method for distilling and the extract application aspect the preparation chemical liver injury of manyflower tickclover herbal extract, but the content that publication relates to is not the whole of manyflower tickclover herbal extract, and its application also only limits to the hepatic injury aspect; The inventor carries out manyflower tickclover herbal extract and should study as further, as enriching and development above-mentioned patent.
Summary of the invention: the objective of the invention is on the basis of above-mentioned invention disclosed the extract of Herba desmodii multifloi and should be as further research, as to above-mentioned invention disclosed enrich and development, make manyflower tickclover herbal extract obtain comprehensive utilization.
Development test proves, manyflower tickclover herbal extract is not only having good effect aspect the control hepatic injury, and aspect the fatty liver that causes in control hyperlipidemia and hyperlipemia good effect arranged also, and particularly the ethyl acetate extract effect is better.
Manyflower tickclover herbal extract of the present invention is meant that with Herba desmodii multifloi plant roots, stem or leaf be raw material; Through 75～95% (V/V) alcohol reflux, reclaim etoh solvent, get ethanol total extract extractum; Total extract extractum is used aqueous dispersion; Use ethyl acetate extraction, reclaim the extractant ethyl acetate, get ethyl acetate extract.
Manyflower tickclover herbal extract of the present invention is used, and ethanol total extract extractum or the ethyl acetate extract that is meant above-mentioned Herba desmodii multifloi plant roots, stem or leaf is used to prevent and treat the application of the medicine of the fatty liver that hyperlipidemia and hyperlipemia cause in preparation.
Ethanol total extract extractum of the present invention or ethyl acetate extract; Can process various preparations, dosage form use; In order to prove that manyflower tickclover herbal extract provided by the invention is to blood fat reducing and the action effect of preventing and treating the hepatic disease that hyperlipemia causes; The present invention adopts the high lipoprotein emulsion of filling stomach to set up mice hyperlipemia liver injury model, and the while, except that blank control group and model group, all the other each groups were irritated the corresponding medicine of stomaches during modeling.Detect TC, TG level and ALT, AST energy value in the mice serum, isolate liver, detect the active and MDA content of SOD in the liver tissue homogenate.Experiment shows: four kinds of extracts of Herba desmodii multifloi (Herba desmodii multifloi leaf ethanol total extract and ethyl acetate extract; Herba desmodii multifloi rhizome ethanol total extract and ethyl acetate extract) all can significantly reduce TG, the TC level of high fat liver damage mice, reduce the activity of ALT, AST, reduce MDA content and SOD activity improving.Wherein best with the accent blood fat and the hepatoprotective effect activity of the ethanol total extract extractum of Herba desmodii multifloi rhizome or Herba desmodii multifloi leaves ethyl acetate extract; Activity is more or less the same under the high low dosage; Good stability; Fat-reducing liver-protecting effect is preferably arranged, can be applied in the medicine aspect of the fatty liver that preparation control hyperlipidemia and hyperlipemia cause.
The application of the ethanol total extract extractum of Herba desmodii multifloi plant roots and stems or Herba desmodii multifloi leaves ethyl acetate extract, also refer to transfer blood fat in preparation, protect the liver and the medicine of the fatty liver that anti peroxidation of lipid and hyperlipidemia cause aspect application.
Effect: the invention provides manyflower tickclover herbal extract and in the medicine of preparation treatment hyperlipemia hepatic injury, use, compare with existing medicine, it is low that it has toxicity; Safe and reliable, drug action is strong, aboundresources; Technology is simple, and the characteristics that preparation cost is low can be processed various dosage forms.
Add 75% (V/V) ethanol after the specific embodiment: embodiment 1, dry Herba desmodii multifloi leaf 1000g pulverize, three times (3 * 3000mL), each 2h merges ethanol extract to reflux, extract,, reclaims solvent, gets ethanol total extract extractum.Ethanol total extract extractum is used aqueous dispersion, and (solvent is reclaimed in 5 * 2000mL) five extractions, gets Herba desmodii multifloi leaves ethyl acetate extract with ethyl acetate.
Add 75% ethanol after dry Herba desmodii multifloi rhizome 1000g pulverizes, three times (3 * 3000mL), each 2h merges ethanol extract to reflux, extract,, reclaims solvent, gets ethanol total extract extractum.Ethanol total extract extractum is used aqueous dispersion, and (solvent is reclaimed in 5 * 2000mL) five extractions, gets Herba desmodii multifloi rhizome ethyl acetate extract with ethyl acetate.
Embodiment 2, present embodiment are the fat-reducing liver-protecting effect experiment of extract.
Principle: normally be called hyperlipemia because lipid metabolism or running make one or more lipids in the blood plasma be higher than unusually.Can show as hypercholesterolemia, hypertriglyceridemia, or both have concurrently.Hyperlipidemia is the main cause that causes the auxotype hepatic injury: triglyceride raises and causes triglyceride acatharsia in the liver in the blood plasma, forms the liver inner lipid and stores up, and causes fatty liver.
Instrument: UV-2000 type ultraviolet-uisible spectrophotometer (but You Ni Shanghai Instr Ltd.); CS-H1 type blender (Beijing rich encourage positive scientific & technical corporation), the biochemical incubator (Shanghai one permanent Science and Technology Ltd.) of LRH-150, Multiskan MK3 ELIASA (thermoelectric Shanghai Instr Ltd.); TGL-16 high speed centrifuge (big instrument plant in Jintan City, Jiangsu); The tissue refiner of 985370-395 type (BIOSREC, Mexico), all size pipettor etc.
Reagent: bifendate drop pill (Zhejiang Prov WanBang Pharmaceutical Co., Ltd, lot number: 080210); Jueming blood-fat-reducing sheet (Changchun people's Pharmaceutical Group Co.,Ltd, lot number: 20071201); The Adeps Sus domestica self-control, cholesterol (purity＞99.5%); No. 3 cholate (the extensive and profound in meaning star in Beijing Bioisystech Co., Ltd, lot numbers: 20080204); Propylthiouracil (Merck schuchardt OHG 85662Hochenbrum, Germany, lot number: 608710); T-CHOL TC, triglyceride TG (North Sea, Shanghai Bioisystech Co., Ltd, lot number: BH-20612); Glutamate pyruvate transaminase ALT, glutamic oxaloacetic transaminase, GOT AST (bio-engineering research institute, lot number 20080807 are built up in Nanjing); Superoxide dismutase SOD (bio-engineering research institute, lot number 20090727 are built up in Nanjing), malonaldehyde MDA (bio-engineering research institute, lot number 20080725 are built up in Nanjing); Coomassie brilliant blue G250 (packing factory of Shanghai chemical reagents corporation, lot number: 20050115)
Method: the configuration of lipomul: get 37 ℃ of thawings of 20g Adeps Sus domestica water-bath, be put in the mortar, in mortar, add the 10g cholesterol, 10g sucrose, the 1g propylthiouracil fully stirs, and dissolving adds tween 80 1mL, and propylene glycol 2mL grinds evenly; Slowly add 10% cholate 20mL, grind emulsifying, adding distil water is to 100mL again.It is subsequent use to be stored in 4 ℃ of refrigerators after preparing.
Each item index determining: select 160 of healthy male mices (body weight 25g ± 2g), be divided into 16 groups at random for use; Be respectively blank control group, model group, bifendate positive controls, jueming blood-fat-reducing sheet positive controls and each administration group, the morning, all the other groups were all irritated stomach Emulsion 0.2mL/10g except that blank control group is irritated stomach (ig) distilled water.Afternoon, blank control group followed model group to irritate the sodium carboxymethyl cellulose (CMC-Na) of stomach (ig) 0.5%, and all the other groups are all irritated the corresponding medicine of stomach (ig).Grouping situation and dosage are seen table 1.After two weeks, water 12h is can't help in fasting before the last administration, and 2h gets blood to mouse orbit after the last administration; The centrifugal 10min of blood 3000r/min; Get its serum, measure serum TG, TC, ALT, AST respectively according to the test kit explanation, the liver that separates liver and the about 300mg of weighing and about 30mg becomes the liver homogenate liquid of 10% (surveying MDA) and 1% (surveying SOD) with 0-3 ℃ of normal saline homogenate; Measure MDA and SOD, measure protein content with Coomassie brilliant blue.
The mensuration of ALT, AST, TG, TC, SOD and MDA is all carried out according to the requirement of test kit; Experimental data
expression, experiment statistics adopt the spss17.0 statistical software to carry out one factor analysis of variance.
The result: (1) manyflower tickclover herbal extract is to the influence of blood fat
Table 1 shows that the TG in the model group serum, TC level are significantly higher than blank control group (P＜0.01, P＜0.001), and the modeling success is described.Compare with model group; TC in the high dose group serum of rhizome total ethanol extract, leaf ethanol total extract and leaves ethyl acetate extract and TG level are starkly lower than model group (P＜0.05; P＜0.01); And rhizome total ethanol extract high dose group TC level is lower than positive control jueming blood-fat-reducing sheet, and three groups TG is all less than positive control jueming blood-fat-reducing sheet.Can find out from the result; The lipid-lowering effect of Herba desmodii multifloi rhizome ethanol total extract is best; Effect for reducing blood fat is suitable with positive control drug jueming blood-fat-reducing sheet; And it is more or less the same at high dose (1000mg/kg) and the TG under low dosage (250mg/kg), TC level, and its good stability is described, has clinical value; Herba desmodii multifloi leaves ethyl acetate extract also all shows lipid-lowering effect preferably when high dose (750mg/kg) and low dosage (187.5mg/kg), also have certain clinical meaning; Though and effect for reducing blood fat is remarkable under the Herba desmodii multifloi leaf ethanol total extract high dose (1000mg/kg); But TG, TC level significantly increase down at low dosage (250mg/kg); And greater than model group; Herba desmodii multifloi rhizome ethyl acetate extract effect for reducing blood fat is not obvious, also shows certain toxicity under the high dose, so both do not have clinical value.
Table 1 manyflower tickclover herbal extract is to the influence (
n=10) of trophism hepatic injury mice T-CHOL, triglyceride
##P＜0.01,
###P＜0.001 is compared with blank control group; P＜0.05,
*P＜0.01 is compared with model group.
(2) manyflower tickclover herbal extract is to the influence of ALT, AST
Table 2 can find out that the ALT in the model group serum, AST are significantly higher than blank control group (P＜0.01, P＜0.001), explain that hyperlipidemia can cause mouse liver injury.Compare with model group, each extract of Herba desmodii multifloi all has ALT and the active ability of AST (P＜0.001, P＜0.01, P＜0.05) in the significant reduction model group mice serum.Compare with positive control drug, the rhizome total ethanol is put forward object height dose groups reduction ALT activity and is superior to bifendate.Can find out from the result; Herba desmodii multifloi rhizome ethanol total extract be have best ALT falls, AST is active; Fall enzymatic activity a little less than bifendate, and it is more or less the same good stability at high dose (1000mg/kg) and the ALT under low dosage (250mg/kg), AST level; Herba desmodii multifloi leaves ethyl acetate extract all has when high dose (750mg/kg) and low dosage (187.5mg/kg) and falls enzymatic activity preferably, also has certain clinical meaning.
Hepatic injury causes hepatic injury mice ALT, AST, MDA, the influence of SOD to table 2 manyflower tickclover herbal extract to trophism
##P＜0.01,
###P＜0.001 is compared with blank control group; P＜0.05.
*P＜0.01 is compared with model group.
(3) manyflower tickclover herbal extract is to the influence of lipid peroxidation
Table 2 shows that the SOD activity in the model group liver homogenate liquid significantly is lower than blank control group (P＜0.001), and MDA content is significantly higher than blank control group (P＜0.01), explains that the hyperlipemia in mice lipid peroxidation strengthens.Compare with model group, rhizome ethanol total extract and ethyl acetate extract high dose group, leaf ethanol total extract and ethyl acetate extract high dose group can significantly reduce MDA content, increased SOD active ((P＜0.01).Wherein the high dose of rhizome ethanol total extract, leaf ethanol total extract and leaves ethyl acetate extract reduces the MDA significance greater than the bifendate positive controls, and the increased SOD significance is suitable with it.It is suitable with the bifendate positive controls with the increased SOD significance that rhizome ethyl acetate extract high dose group reduces MDA.The result shows that the high low dose group of Herba desmodii multifloi rhizome ethanol total extract and Herba desmodii multifloi leaves ethyl acetate extract all has good reduction MDA and increased SOD is active, explains that both antioxidant effects are good, good stability.
In sum, Herba desmodii multifloi rhizome ethanol total extract and leaves ethyl acetate extract have transfer the most significantly blood fat (TG, TC), protect the liver (ALT, AST) and anti peroxidation of lipid (MDA, SOD) effect, both all can be used for treating the hepatic injury that hyperlipidemia causes.
The preparation of embodiment 3, manyflower tickclover herbal extract tablet:
Method for preparing: get above-mentioned Herba desmodii multifloi rhizome ethanol total extract 100g, the methanol of adding an amount of 80% is made the wetting agent mixing and is processed soft material behind adding starch 250g, the arabic gum 15g, under 60 ℃ of temperature, is dried to water content below 3%; Then the thing of oven dry was pulverized 14 mesh sieves, add magnesium stearate 5g and Pulvis Talci 20g mixing at last after, cross 12 mesh sieves; Granulate; Tabletting, quality inspection, coating are processed 1000.
It should be noted last that: above embodiment is only in order to explanation; And unrestricted technical scheme of the present invention; Although the present invention is specified with reference to the foregoing description; Those of ordinary skill in the art is to be understood that: still can make amendment or be equal to replacement the present invention, and not break away from any modification or the local replacement of the spirit and scope of the present invention, it all should be encompassed in the middle of the claim scope of the present invention.
Claims (3)
1. a manyflower tickclover herbal extract is characterized in that with Herba desmodii multifloi plant drying leaf be raw material, through 75～95% alcohol reflux, reclaims etoh solvent, gets ethanol total extract extractum; Total extract extractum is used aqueous dispersion, uses ethyl acetate extraction, reclaims the extractant ethyl acetate, gets ethyl acetate extract.
2. the application of manyflower tickclover herbal extract, described manyflower tickclover herbal extract are to be raw material with Herba desmodii multifloi plant drying root, stem or leaf, through 75～95% alcohol reflux, reclaim etoh solvent, ethanol total extract extractum; Total extract extractum is used aqueous dispersion, uses ethyl acetate extraction, reclaims the extractant ethyl acetate, gets ethyl acetate extract; Ethanol total extract extractum or the Herba desmodii multifloi leaves ethyl acetate extract that it is characterized in that referring to Herba desmodii multifloi plant roots, stem is used to prevent and treat the application aspect the medicine of the fatty liver that hyperlipidemia and hyperlipemia cause in preparation.
3. the application of manyflower tickclover herbal extract according to claim 2, ethanol total extract extractum or the Herba desmodii multifloi leaves ethyl acetate extract that it is characterized in that Herba desmodii multifloi plant roots, stem the preparation blood fat reducing, protect the liver and the medicine of anti peroxidation of lipid aspect application.
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