CN1887301A - Application of spanishneedles herb flavonid in preparing medicine for preventing and treating viral hepatitis - Google Patents

Application of spanishneedles herb flavonid in preparing medicine for preventing and treating viral hepatitis Download PDF

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CN1887301A
CN1887301A CNA2005100973384A CN200510097338A CN1887301A CN 1887301 A CN1887301 A CN 1887301A CN A2005100973384 A CNA2005100973384 A CN A2005100973384A CN 200510097338 A CN200510097338 A CN 200510097338A CN 1887301 A CN1887301 A CN 1887301A
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medicine
viral hepatitis
general flavone
hepatitis
sticktight general
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CN100486599C (en
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陈飞虎
李俊
钟明媚
吕雄文
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Anhui Medical University
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Anhui Medical University
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Abstract

The present invention belongs to the field of Chinese medicine technology, relates to medicine for treating viral hepatitis, and is especially application of sponish needles herb flavonid in preparing medicine for preventing and treating viral hepatitis. The medicine of the present invention has the functions of clearing away heat and toxic material, promoting blood circulation to disperse blood clots, soothing liver and invigorating spleen, and is suitable for various kinds of viral hepatitis. The medicine of the present invention has determined curative effect and low toxic side effect.

Description

Sticktight general flavone is prevented and treated application in the viral hepatitis medicine in preparation
Technical field
The present invention relates to the purposes of Sticktight general flavone, the definite Sticktight general flavone of saying so is prevented and treated purposes in the viral hepatitis medicine in preparation.
Background technology
Herba Bidentis Bipinnatae has another name called Herba Bidentis Bipinnatae, terrible wooden fork grass, little Bidens bipinnata, sticking body grass etc., its medicinal history is long, the beginning is stated from Tang's CHEN Zang-Qi supplement to the Herbal, meaning: " terrible hairpin grass, bitter in the mouth is flat, nontoxic, main Serpentis and Aranea sting, broken Fu of pestle it, also pestle strand juice clothes.There is fork at Radix Rehmanniae side, and square-stem, son are made the hairpin foot, people such as clothing pin, and what exhaled in the north is Bidens bipinnata.But " all medications of Herba Bidentis Bipinnatae class medical material herb, bitter in the mouth is flat and nontoxic, has heat-clearing and toxic substances removing, and effects such as dissipating blood stasis for subsidence of swelling cure mainly malaria, diarrhoea, dysentery, hepatitis, acute nephritis, stomachache, dysphagic, acute appendicitis, laryngopharynx swelling and pain, traumatic injury, snake bite and insect sting etc.In order better to be familiar with and to understand this natural resources, better develop and use, existing that the research and development review of Present Situation of Herba Bidentis Bipinnatae is as follows.
Herba Bidentis Bipinnatae is that Compositae (Compositae) tubular flower subfamily Bidens (Bidens L.) plant is an annual herb, its herb resource is very abundant, mainly be distributed in ground such as China northeast, Hebei, Shandong, Shanxi, Henan, Jiangsu, Yunnan, Sichuan, Guangdong and Guangxi Provinces, be born in waterside, riverbank, wetland, wasteland and hillside etc. and locate, very easily gather.The chemical constituent of Herba Bidentis Bipinnatae:
The contained chemical constituent kind of Herba Bidentis Bipinnatae is more, and bibliographical information once separated from this plant and obtains volatile oil, sterol, chalcone derivative, aurones, polyacetylene class etc., and its active component is based on polyacetylene class and flavones ingredient.Isolated multiple polyacetylene constituents.The n-butyl alcohol of ethanol extraction partly identifies 4 flavone components, is hyperin (hyperoside), okanin (okanin), isookanin-7-O-glycoside and maritimetin (maritimein).In each position of Herba Bidentis Bipinnatae there be than big-difference flavones ingredient, at flowering fruit bearing stage, and general flavone content higher (being respectively 6.67% and 6.68%) in leaf and the inflorescence, content lower (0.89%) in the stem branch.In the leaf isolation identification 21 kinds of volatile ingredients, be mainly fatty alkane and fatty acid, contain a spot of aldehyde, ketone alkene, nitrile, amine and furans composition in addition; Wherein fats compound accounts for significant proportion.Isolated 4 kinds of liposoluble ingredients through physics and chemistry test and spectrum analysis, are accredited as salicylic acid, protocatechuic acid, gallic acid, fatty acid compound respectively.Also have steroid compound such as spinach sterol, daucosterol, sitosterol, stigmasterol-3-O-β-D-glucoside.
Herba Bidentis Bipinnatae contains rich in amino acid, as 17 seed amino acids such as aspartic acid, threonine, serine, glutamic acid, alanine, valine, isoleucine, proline, tyrosine, lysines, wherein 8 kinds is essential amino acid, gross protein 157.89g/kg, total free amino acid content reaches 139.89g/kg, TEAA 57.09g/kg.Amino acid contained and the traditional rare medicinal herbs Cordyceps hawkesii Gary of Herba Bidentis Bipinnatae, Cordyceps are compared, many cystine on kind, and proline, glutamic acid are higher than both on content, and other amino acid contents are suitable
Also contain 11 kinds of micro elements needed by human in addition in the Herba Bidentis Bipinnatae, measurement result shows, Mg content is the highest, P, Fe, Ca content are higher, Cu, Zn take second place, Mn, Co, Ni, Mo, Cr content are minimum, contain abundant P in the Herba Bidentis Bipinnatae, are that it regulates immanent cause that nerve, hypnosis, calmness etc. act on.
Vitamin also has distribution in Herba Bidentis Bipinnatae, as vitamin B 1, vitamin B 2, beta-carotene.
The Herba Bidentis Bipinnatae herb resource is abundant, and medicinal history is long, and Radix Codonopsis Cardiophyllae's title is arranged in folk remedy.Its biological activity is various, and action temperature and lasting, has higher medical value, especially digestive system, central nervous system and cardiovascular system is shown stronger pharmacological action.In recent years the Herba Bidentis Bipinnatae medical material some developments and utilizations have been carried out, domestic Herba Bidentis Bipinnatae product has the year of a pleasing sheet, Herba Bidentis Bipinnatae oral liquid, Herba Bidentis Bipinnatae health instant powder etc. at present, but also only limit to prevent and treat cardiovascular disease such as hypertension, yet there are no Sticktight general flavone at the report aspect the antiviral hepatitis.
Summary of the invention
The new purposes that the purpose of this invention is to provide Sticktight general flavone, i.e. new application in pharmacy.
In fact, the invention provides the application of Sticktight general flavone in the medicine of preparation prevention or treatment viral hepatitis.
Viral hepatitis is the infectious disease that is caused by hepatitis virus, generally acknowledge so far and can be divided into first, second, third, fourth, penta 5 kinds of types, its infectiousness is strong, the route of transmission is complicated, the sickness rate height, and a large amount of virus carriers is arranged, this class crowd is potential morbidity object, and with liver cirrhosis, hepatocarcinoma substantial connection is arranged.Viral hepatitis can will be divided into high, medium and low popular district all over the world in the distribution on global difference according to popularity.China is the high epidemic regions of first, hepatitis B, and it is popular that hepatitis C is moderate.Hepatitis is the popular serious harm people's of hepatitis B health especially, and socio-economic development is had the restrictive function that can not be ignored, and the problem of preventing and treating of hepatitis has become the common important topic of paying close attention to of whole world medical circle.
At present clinical antihepatitis drug commonly used can be divided three classes substantially: 1. antiviral agents; 2. immunomodulator; 3. hepatoprotective etc.In many therapeutic schemes, implementing antiviral therapy at the cause of disease is an important directions.Interferon is at first to obtain the extensively antiviral drugs of approval, also is to treat the hepatitis B choice drug at present, but only 30%～40% hepatitis B patient is had long-term effect.Nucleoside analog such as lamivudine (Lamividine) though etc. have a curative effect preferably, relapse rate is quite high after the drug withdrawal.The therapeutic scheme that some are new such as antisensenucleic acids, ribozyme etc., since a series of major issues, as the safety of virus expression carrier, the introduction method of gene transfer target cell, expression that how to stop quiding gene after the treatment etc. is still unresolved, thereby can only rest in the experiment.Generally speaking, lack the medicine of hepatitis virus resisting safely and effectively at present.
And the treatment strategy of modern medicine is turning to the adjusting countermeasure from allopathy, in changing body, promotes the research of body defence capability from antiviral research trend.This developing direction, the advantage of Chinese medicine and characteristics just.According to Traditional Chinese medical theory, viral hepatitis is that epidemic disease caused by damp-heat pathogen poison is invaded liver and gall, causes function of liver and spleen damage etc. relevant, answers nourishing the liver and kidney, clearing away heat-damp and promoting diuresis, invigorating the spleen for dissipating phlegm, detoxifcation to fall enzyme etc. so treat primary disease.The present invention follows this Therapeutic Principle, based on the long-term treatment experience, finds that Herba Bidentis Bipinnatae has heat-clearing and toxic substances removing, and effects such as dissipating blood stasis for subsidence of swelling have good therapeutical effect to viral hepatitis.For being developed to, it provides the foundation that is of great rarity to the active drug of preventing and treating viral hepatitis.We extract its effective site total flavones, confirm that by experiment in vivo and vitro it has significant antiviral hepatitis effect.
In order to understand essence of the present invention better, will its application in preventing and treating the viral hepatitis medicine be described with the pharmacological evaluation and the result of Sticktight general flavone below.
It is as follows that Sticktight general flavone is prevented and treated the viral hepatitis test:
(1) preparation of Sticktight general flavone
Get the Herba Bidentis Bipinnatae crude drug and pulverize, the alcohol reflux of 40-80% 2-3 time, extracting solution merges, and reclaims ethanol, and concentrating under reduced pressure gets extractum.Extractum is water-soluble, leaves standstill, filter, and resin column on the clear liquor, 40-95% ethanol elution part is collected in the washing back, reclaims ethanol, and concentrating under reduced pressure is drying to obtain extract, and described extract shows Sticktight general flavone.Sticktight general flavone is the yellow green powder, has fragranced, easily deliquescence; Its main component general flavone content is 20-60%.Extract can add corresponding adjuvant, makes tablet, capsule, slow releasing tablet, drop pill, electuary, microgranule, and is further purified and makes injection.
(2) experiment
1. Sticktight general flavone causes the protective effect of chmice acute chemical liver injury to carbon tetrachloride
(1) material and reagent: Sticktight general flavone; 0.1%CCl 4Bifendate; ALT, the AST test kit: biotech company is built up in Nanjing; 60 of Male Kunming strain mice, body weight 18-22g is provided by Medical University Of Anhui zoopery center.
(2) method: 60 of normal mouses divide 6 groups at random, gastric infusion (Sticktight general flavone 80mg/kg, 160mg/kg, 320mg/kg, positive drug control group bifendate 100mg/kg) or equal-volume normal saline (normal control, model), every day 2 times, continuous 7d, 2h after the last administration, normal control group ip10ml/kg Oleum Arachidis hypogaeae semen, all the other 5 groups of equal ip0.1%CCl 410ml/kg from mice posterior orbit vein treating the preponderant disease instead of the secondary disease blood, measures Serum ALT behind the fasting 16h, the AST activity, and get the same position of liver lobus sinister, fixing, do pathological section, carry out hepatic pathology histology HE chromoscopy, the result with hepatic pathology degree of injury scoring under the light microscopic and pathology photo as standard.
Liver tissue lesions's grading method: light microscopic is checked down, is divided into 5 grades by the hepatic necrosis degree:
0 grade, normal or liver has slight degeneration, and hepatocyte does not have necrosis;
The I level, spotty necrosis, necrosis accounts for lobules of liver＜1/4;
The II level, the necrosis of some kitchen range shape, necrosis accounts for lobules of liver 1/4-1/3;
The III level, the necrosis of kitchen range shape, necrosis accounts for lobules of liver＜1/2;
The IV level, lamellar necrosis, necrosis account for lobules of liver＞1/2.
(3) result: table 1 is the result show, CCl 4ALT, AST activity in the remarkable rising mice serum, and Sticktight general flavone significantly reduces CCl 4Cause ALT too high in the acute chemical hepatic injury mice serum, AST activity.
Table 1 Sticktight general flavone is to the influence of ALT, AST in the mice serum (n=10, x ± s)
Group Dosage/mgkg -1 ALT/U·L -1 AST/U·L -1
Matched group model group positive drug Sticktight general flavone - - 100 320 160 80 46.13±6.76 144.73±12.11** 46.61±12.51## 57.80±30.22## 75.96±32.58## 105.72±30.00## 45.21±16.12 93.24±36.11** 62.05±14.09 54.79±14.82## 37.96±14.10## 79.85±26.75
* p＜0.01 is compared with matched group; ##p＜0.01 is compared with model group
Table 2 result shows that necrosis in different degree appears in the visible inflammatory cell infiltration of model group hepatic tissue.Sticktight general flavone can obviously alleviate liver inflammatory cell infiltration and necrosis.
Table 2 Sticktight general flavone is to CCl 4Cause the influence of acute chemical hepatic injury mice histopathology
Group Dosage mg/kg Classification
0 I II III IV
Normal group - 10 0 0 0 0
Model group - 3 4 2 2 0
The positive drug group 100 11 0 0 0 0
The Sticktight general flavone group 320 10 2 0 0 0
160 12 0 0 0 0
80 0 2 3 3 4
1, Sticktight general flavone anti-hepatitis virus test
(1) anti-hepatitis virus in vivo test:
Guangzhou sheldrake with infected duck hepatitis B virus (DHBV) is an experimental animal model.50 of 1 age in days Guangzhou sheldrakes are through lower limb intravenous injection Shanghai sheldrake DHBV-DNA positive serum, every 0.2ml.Infected 13 days after lower limb shin vein is got blood, survey serum DHBV-DNA total positives.Be divided into 5 groups at random, the Sticktight general flavone group is respectively with 100mg/kg.d, 200mg/kg.d and 400mg/kg.d administration, and positive control drug acycloguanosine (ACV) is with the 100g/kg.d administration, and the virus control group is irritated stomach, every day 2 times, totally 14 days with the equivalent normal saline.(T before administration 0), the 7th day (T of administration 7), 14 days (T 14) get blood from duck lower limb shin vein, separation of serum, the level of DHBV-DNA the results are shown in Table 3 during the survey Sanguis Anas domestica was clear.
Table 3 is respectively organized in the duck body DHBV-DNA relatively (OD value, x ± s)
Group Dosage (mg/kg.d) DHBV-DNA
T 0 T 7 T 14
Virus control ACV group Sticktight general flavone group - 100 100 200 400 1.11±0.27 1.41±0.26 1.50±0.41 1.70±0.13 1.76±0.22 1.06±0.25 0.06±0.02**## 0.98±0.60 0.84±0.56* 0.71±0.43* 1.01±0.26 0.05±0.04**## 0.82±0.63* 0.59±0.56**# 0.32±0.20**#
Annotate: with comparison before the administration of this group, * P＜0.05, * * P＜0.01; Compare #P＜0.05, ##P＜0.01 same period with the virus control group
Table 3 result shows that Sticktight general flavone has remarkable effect to the rising that the reduction dhbv dna infects the DHBV-DNA that causes, and is wherein remarkable with the effect of 400mg/kg.d group.
(2) anti-hepatitis virus in vitro tests
Adopt HepG 22.2.15 cell is tested the anti-HBV effect of medicine.The 2.2.15 cell recovery is cultivated back piping and druming gently with 0.06% trypsinization and with the 2.2.15 cell be dispersed into the individual cells suspension, being made into cell concentration with the DMEM culture fluid that contains 10% hyclone is 3 * 10 5The cell suspension of individual/ml is planted respectively in 96 orifice plates by the 0.1ml/ hole, puts 37 ℃, saturated humidity, 5%CO 2Cultivate in the incubator, add after 2 days with the pastille cultivation and keep liquid 0.1ml/ hole.The medicine Sticktight general flavone with the culture fluid that contains 2% hyclone be mixed with 10,5,2.5,1.25,0.63, six kinds of concentration of 0.31mg/ml, every kind of drug level all adds 4 holes, changed liquid 1 time in per 4 days, and measured HBsAg and HBeAg with the ELISA method in cultivating the 12nd day gentle aspiration cultured cell supernatant.The results are shown in Table 4:
Table 4 Sticktight general flavone is to the inhibitory action of 2.2.15 emiocytosis HBsAg, HBeAg
Group Concentration (mg/ml) HBsAg HBeAg
A( x±s) Suppression ratio (%) A( x±s) Suppression ratio (%)
Sticktight general flavone cell contrast blank 10 5 2.5 1.25 0.63 0.31 0.051±0.01 0.06±0.02 0.05±O.03 0.09±0.OO 0.10±0.03 0.11±0.04 0.76±0.07 0.039 105 100 105 83 78 72 0.10±0.00 0.16±0.02 0.22±0.07 0.24±0.05 0.21±0.07 0.49±0.12 0.64±0.19 0.023 88 78 68 65 70 24
Table 4 result show Sticktight general flavone to the suppression ratio of HBsAg all more than 70%, comparatively obvious when 0.63mg/ml is above to the antigenic inhibitory action of HBeAg.This result shows that Sticktight general flavone can significantly suppress 2.2.15 emiocytosis HBsAg, HBeAg, has the effect of stronger vitro inhibition 2.2.15 cellular expression antigen of hepatitis B virus, and the prompting Sticktight general flavone effectively suppresses the effect of HBV in vitro tests.
Above-mentioned experimental result shows that the present invention has the advantage of several aspects:
1, the present invention has excavated the new prospect in medicine of Sticktight general flavone, has opened up a new medicinal field;
2, Sticktight general flavone of the present invention adopts oral administration, taking convenience, and toxic and side effects is little, takes medicine for a long time and does not have harm;
3, the former medicine resource of Sticktight general flavone of the present invention is very abundant, has good society and economic benefit;
4, Sticktight general flavone of the present invention is applicable to viral hepatitis, acts on a plurality of links, determined curative effect;
5, the present invention provides strong basis for Herba Bidentis Bipinnatae being developed further into new drug.
The specific embodiment
Below in conjunction with embodiment the present invention is done to describe further.
Embodiment 1:
Take by weighing Herba Bidentis Bipinnatae crude drug 10kg, pulverize the back and add 80% ethanol 100L, hot reflux is extracted 2 times, each 1h, and merge extractive liquid, reclaims ethanol, and concentrating under reduced pressure gets extractum.Extractum is water-soluble, leaves standstill, filter, and macroporous resin column on the clear liquor, 80% ethanol elution part is collected in the washing back.Reclaim ethanol, concentrating under reduced pressure is drying to obtain extract, and extract is a Sticktight general flavone.Sticktight general flavone is that yellow green is Powdered, has fragranced, easily deliquescence; Its main component general flavone content is 20-60%.Extract can add corresponding adjuvant, makes tablet, capsule, slow releasing tablet, drop pill, electuary, microgranule, and is further purified and makes injection.
The clinical recommended dose of Sticktight general flavone: 300-1200mg/ days, oral, three times on the one.

Claims (1)

1, Sticktight general flavone is prevented and treated application in the viral hepatitis medicine in preparation, it is characterized in that: Sticktight general flavone is prevented and treated application in the viral hepatitis medicine in preparation.
CN 200510097338 2005-12-30 2005-12-30 Application of spanishneedles herb flavonid in preparing medicine for preventing and treating viral hepatitis Expired - Fee Related CN100486599C (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102018741A (en) * 2010-12-10 2011-04-20 安徽医科大学 Spanishneedles herb total flavone extract as well as preparation technology and preparation thereof
CN102716166A (en) * 2012-07-10 2012-10-10 无锡市第三人民医院 Method for extracting total flavonoids of bidens bipinnata and application of total flavonoids of bidens bipinnata (TFB) in preparation of medicine for treating rheumatoid arthritis

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102018741A (en) * 2010-12-10 2011-04-20 安徽医科大学 Spanishneedles herb total flavone extract as well as preparation technology and preparation thereof
CN102018741B (en) * 2010-12-10 2012-11-07 安徽医科大学 Spanishneedles herb total flavone extract as well as preparation technology and preparation thereof
CN102716166A (en) * 2012-07-10 2012-10-10 无锡市第三人民医院 Method for extracting total flavonoids of bidens bipinnata and application of total flavonoids of bidens bipinnata (TFB) in preparation of medicine for treating rheumatoid arthritis

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