CN104784186A - Application of asiatic acid in preparation of medicine for preventing hepatitis virus - Google Patents
Application of asiatic acid in preparation of medicine for preventing hepatitis virus Download PDFInfo
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- CN104784186A CN104784186A CN201510225621.4A CN201510225621A CN104784186A CN 104784186 A CN104784186 A CN 104784186A CN 201510225621 A CN201510225621 A CN 201510225621A CN 104784186 A CN104784186 A CN 104784186A
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Abstract
The invention discloses application of asiatic acid in preparation of a medicine for preventing hepatitis virus. The chemical structure of the asiatic acid is as shown in formula I. Through in vitro building a hepatitis c virus (HCV cc) model derived from cell culture, and using an HCV susceptible cell human liver cancer Huh7 cell line, an in-vitro experiment on the anti-HCV activity of the asiatic acid is performed. The result shows that the asiatic acid has obvious anti-HCV activity, particularly is capable of inhibiting the HCV invading the target cell and can be used for preparing the anti-hepatitis c virus medicine. A novel path for the asiatic acid is provided and a novel source for finding the anti-hepatitis c virus medicine is also provided.
Description
Technical field
The present invention relates to medical art, specifically, is the application of asiatic acid in preparation control hepatitis virus medicament.
Background technology
Asiatic acid (asiatic acid, AA) is a kind of Ursane Pentacyclic triterpenic acid, is present in various plants, as samphire Herba Centellae.Asiatic acid structure is as follows:
The conventional Chinese medicine that Herba Centellae is recorded for " Chinese Pharmacopoeia " is the dry herb of Umbelliferae Umbelliferae Centella Centella plant Centella asiatica, within 1977 years, begins to be loaded into Pharmacopoeia of People's Republic of China.About there is Centella CentellaL. plant 20 kinds in the whole world, is mainly distributed in tropical and subtropical region.Asiatic acid is the main component of asiatic centella triterpenoid extract, and also can be obtained by asiaticoside hydrolysis, can also extract from other natural products many and obtain, therefore resource is very abundant.Asiatic acid has pharmacological action widely; such as antiinflammatory skin protection, protect the liver and protect lung, antidepressant, blood sugar lowering, blood fat reducing, neuroprotective, antitumor etc. (see the Central China University of Science and Technology; Ph.D. Dissertation; the material base research of cool water harm, A.chinensis Planch. root, 144 pages) effect.Chinese patent literature CN200610118041.6 discloses the application of asiatic acid at aspect of resisting pulmonary fibrosis.Chinese patent literature CN20061012256.4 discloses asiatic acid and is preparing the new opplication in medicine, is particularly preparing the purposes in anti-hepatic fibrosis medicines.But about the effect of asiatic acid anti-hepatitis c virus, yet there are no report.
Summary of the invention
The object of the invention is for deficiency of the prior art, the application of asiatic acid in preparation control hepatitis virus medicament is provided.
Of the present invention again one object be provide asiatic acid preparing the application in the food that protects the liver and health product.
For achieving the above object, the technical scheme that the present invention takes is:
The application of asiatic acid in preparation control hepatitis virus medicament, described asiatic acid chemical structural formula is as follows:
The application in anti hepatitis C virus drug prepared by described asiatic acid.
Described asiatic acid suppresses the application in the medicine of HCV invasion target cell in preparation.
For realizing above-mentioned second object, the technical scheme that the present invention takes is:
The application in the food and health product protected the liver prepared by asiatic acid.
The invention has the advantages that:
The present invention provides one to prepare the purposes of anti-hepatitis c virus (HCV) medicine for Pentacyclic triterpenic acid asiatic acid, by the hepatitis C virus (HCVcc) that the cell culture of external foundation is originated, utilize HCV easy infection cell people hepatocarcinoma Huh7 cell line, experiment in vitro has been carried out to the anti-HCV activity of Pentacyclic triterpenic acid asiatic acid.Found that, asiatic acid has significant anti-HCV activity, and HCV especially can be suppressed to invade target cell, can be used for the medicine preparing anti-third type virus.The present invention is not only Pentacyclic triterpenic acid and provides novelty teabag, provides new source for finding anti hepatitis C virus drug yet.
Detailed description of the invention
Below in conjunction with embodiment, detailed description of the invention provided by the invention is elaborated.
The anti-third type hepatovirus experiment of embodiment 1 asiatic acid
One, Experimental agents, reagent and material
1. compound: asiatic acid.
2. cell line Huh7, human hepatoma cell strain (refers to: Yimin Tong, Yongzhe Zhu, Xueshan Xia, Yuan Liu, et al.Tupaia CD81, SR-BI, Claudin-1, and Occludin Support Hepatitis CVirus Infection, JOURNAL OF VIROLOGY, 2011; 85 (6): 2793 – 2802; Jin Zhong, Pablo Gastaminza, Guofeng Cheng, et al.Robust hepatitis C virus infection in vitro, PNAS, 2005; 102 (26): 9294-9299).
3. cell line 293T, human embryonic kidney cell line (is referred to: Yimin Tong, Yongzhe Zhu, Xueshan Xia, Yuan Liu, et al.Tupaia CD81, SR-BI, Claudin-1, and Occludin Support Hepatitis CVirus Infection, JOURNAL OF VIROLOGY.2011; 85 (6): 2793 – 2802; Graham, F.L., Smiley, J., Russell, W.C. & Nairn, R.J.Gen.Virol 1977; 36:59 – 74).
4. cell culture fluid, preparation, it adjusts pH to 7.4 containing 10% hyclone, 0.03% glutamine, non essential amino acid, ampicillin and streptomycin 100U/ml.
5. cell dissociation buffer, preparation, it, containing 0.25% trypsin, is prepared with phosphate buffer.
6.HCVcc: the infectious hepatitis C virus of cell culture (refers to: Yimin Tong, Yongzhe Zhu, Xueshan Xia, Yuan Liu, et al.Tupaia CD81, SR-BI, Claudin-1, and Occludin SupportHepatitis C Virus Infection, JOURNAL OF VIROLOGY, Mar.2011; 85 (6): 2793 – 2802; Jin Zhong, Pablo Gastaminza, Guofeng Cheng, et al.Robust hepatitis C virusinfection in vitro, PNAS, 2005; 102 (26): 9294-9299).
Two, experimental technique:
(1) preparation of HCVcc
1. virus amplification
J6, JFH-1 are fitted together to HCVcc (10
5ffu/ml), get 50 μ l and infect Huh 7.5 cell being inoculated in 24 orifice plates, next day changes liquid, subsequently according to cell density Secondary Culture, observation of cell growth conditions, after cytopathic effect (CPE) appearance that viral fast breeding causes, collect the culture supernatant of 7th ~ 20 days, get and measure for virus titer in right amount, after all the other 8,000rpm centrifugal 5min abandon cell debris subpackage be stored in-70 DEG C for subsequent use.
2. titration of virus
With blank Huh 7.5 (1 × 10
4cells/ hole), after 12h, abandon culture supernatant, every hole adds the HCVcc supernatant of 100 μ l through 10 times of gradient dilutions, hatch 5h altogether, renew the full culture fluid of fresh DMEM and continue to cultivate 72h, row immuno-fluorescence assay HCV positive cell, the HCV antibody positive patients serum that primary antibodie is diluted with 1:100, two resist the FITC labelling goat anti-human igg for 1:100 dilution.At fluorescence microscopy Microscopic observation luminescent cell, and to record in last hole that can be observed gfp positive cell gfp positive cell number and dilute gradient accordingly, calculate focus forming unit/ml (ffu/ml) numerical value, represent HCVcc titre with this.
(2) the infective detection of HCVcc
Get the Huh7 cell being in exponential phase, adjustment cell concentration is 1 × 10
5individual/ml, gets 100 μ l kind 96 orifice plates; Cultivate after 24h and add compound and HCVcc, asiatic acid is by 10 μ g/ml and the preparation of 20 μ g/ml concentration, and 37 degree cultivations remove compound after 4 hours and HCVcc mixes liquid, change culture medium and continue cultivation; Carry out Immunofluorescence test after 48h, under fluorescence microscope, read each hole HCVcc positive colony number.
(3) result (see table 1)
Table 1 asiatic acid is to the inhibit activities (%) of cell culture hepatitis C virus (HCVcc)
Above-mentioned experimental result shows, asiatic acid has significant antihepatitis C virus activity.Therefore, can be used for the medicine preparing anti-hepatitis c virus and the medicine protected the liver.
The hepatitis C virus (HCVcc) that the present invention is originated by the cell culture of external foundation, utilizes HCV easy infection cell people hepatocarcinoma Huh7 cell line, has carried out experiment in vitro to the anti-HCV activity of pentacyclic triterpene acid compound asiatic acid.Found that, asiatic acid has significant anti-HCV activity, and HCV especially can be suppressed to invade target cell.Therefore, asiatic acid can be used for the medicine preparing anti-hepatitis C virus (HCV).
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the prerequisite not departing from the inventive method; can also make some improvement and supplement, these improve and supplement and also should be considered as protection scope of the present invention.
Claims (4)
1. the application of asiatic acid in preparation control hepatitis virus medicament, it is characterized in that, described asiatic acid chemical structural formula is as follows:
2. application according to claim 1, is characterized in that, the application in anti hepatitis C virus drug prepared by described asiatic acid.
3. application according to claim 1, is characterized in that, described asiatic acid suppresses the application in the medicine of HCV invasion target cell in preparation.
4. the application in the food and health product protected the liver prepared by asiatic acid.
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| CN201510225621.4A CN104784186B (en) | 2015-05-06 | 2015-05-06 | Asiatic acid is preparing the application in preventing hepatitis virus medicament |
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| CN201510225621.4A CN104784186B (en) | 2015-05-06 | 2015-05-06 | Asiatic acid is preparing the application in preventing hepatitis virus medicament |
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| CN104784186B CN104784186B (en) | 2018-08-28 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107056874A (en) * | 2016-12-28 | 2017-08-18 | 福建广生堂药业股份有限公司 | A kind of compound of asiatic acid tenofovir dipivoxil and preparation method thereof |
| CN114796233A (en) * | 2022-06-17 | 2022-07-29 | 重庆医科大学国际体外诊断研究院 | Application of asiatic acid in preparing medicine for treating hepatitis B |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1943579A (en) * | 2006-09-30 | 2007-04-11 | 佛山市第一人民医院 | Use of asiatic acid in preparing medicine |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1943579A (en) * | 2006-09-30 | 2007-04-11 | 佛山市第一人民医院 | Use of asiatic acid in preparing medicine |
Non-Patent Citations (1)
| Title |
|---|
| 孙瑞娜等: "女贞子提取物主要活性组分及其抗丙型肝炎病毒复制活性分析", 《中国生物化学与分子生物学报》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107056874A (en) * | 2016-12-28 | 2017-08-18 | 福建广生堂药业股份有限公司 | A kind of compound of asiatic acid tenofovir dipivoxil and preparation method thereof |
| CN114796233A (en) * | 2022-06-17 | 2022-07-29 | 重庆医科大学国际体外诊断研究院 | Application of asiatic acid in preparing medicine for treating hepatitis B |
| WO2023241179A1 (en) * | 2022-06-17 | 2023-12-21 | 重庆医科大学检验医学院 | Use of asiatic acid in preparation of medicament for treating hepatitis b |
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Granted publication date: 20180828 Termination date: 20210506 |