The Study On Anti-liver Injury And Anti-HBV Of Clinacanthus Nutans

ObjectiveIn this paper by observe relate index we be able to evaluate the protective effect of Clinacanthus nutans（CN）on liver injury which is induced by carbon tetrachloride（CC14）and alpha-naphthylisothiocyanate（ANIT）in vivo respectively,also is induced by carbon tetrachloride（CC14）to L-02 cells in vitro.Then,we can obtain secretion of antigen by HepG2.2.15 cell model,in the meantime,it can evaluate the effect of drug-containing serum in vitro on the inhibition of hepatitis B virus（HBV）.Finally,by discuss the anti-liver injury and anti-HBV effect of CN,we can provide the experimental basis for the further development and utilization of CN.Methods（1）Protective effect of Clinacanthus nutans on liver injured①The mice were randomly divided into 9 groups（n=10）,namely:normal group,CC14 model group,positive control group,CN fresh juice groups（78g · kg-1,156g · kg-1）,CN fresh groups（78g · kg-1,156g · kg-1）,CN dry groups（15.6g · kg-1,31.2g · kg-1）.Normal group and CC14 model group were given distilled water,while other groups were given relevant medicines intragastrically once a day for consecutive 15 days.Except the normal group,other groups were given 0.1%CCl₄（diluted with peanut oil solution）intraperitoneal injection,the related indexs were determined and compared between groups and observed the liver biopsy pathology.②The mice were randomly divided into 7 groups（n=10）,namely:normal group,ANIT model group,positive contorl group,CN fresh juice groups（78g·kg-1、156g·kg-1）,CN fresh groups（78g·kg-1、156g·kg-1）.Normal group and ANIT model group were given distilled water,while other groups were given relevant medicines intragastrically once a day for consecutive 15 days.Except the normal group,other groups were given 2%ANIT（diluted with peanut oil solution）intragastric administration.the related indexs were determined and compared between groups and to observe the liver biopsy pathology.③ The SD rat were randomly divided into 3 groups,namely:normal group,positive contorl group,fresh juice’ s CN group,all groups were given relevant medicines intragastrically twice a day for consecutive 3 days.While blood was collected from the abdominal aorta,and then collecte the serum after processing for storing and using.Selected the logarithmic growth phase of L-02 liver cells were divided into 6 groups,namely:cell control group,model contorl group,positive contorl group,CN serum containing groups（25%,50%,100%）.Except the cell control group,other groups were given CCl₄ to intervention after 6 hours termination of molding.All groups were given relevant serum containing medicines 2 hours after termination of the intervention,and to detect the growth and the survival rate of L-02 cells by MTT assay.In addition,to detect the content of GPT,GOT,SOD,MDA by use hepatic cell homogenate enzyme mark instrument.（2）Preliminary study of the serum-containing Cl inacanthus nutans anti-HBV in vitroThe SD rat were randomly divided into 2 groups,namely:normal contorl group,fresh juice’ s CN group,all groups were given relevant medicines intragastrically twice a day for consecutive 3 days.While blood was collected from the abdominal aorta,and then collecte the serum after processing for storing and using.Selected the logarithmic growth phase of HepG2.2.15 cells were seeded in 96-well cell culture cluster,under the condition of 37℃,5%CO2 were cultured to detect the Iconic antigen content of HBsAg,HBeAg in HepG2.2.15 cells supernatant.cells were seeded in plate,cells were divided into 4 groups,namely:cell control group,CN serum containing groups（25%,50%,100%）.the cell physiological status were observed at any time,to detect the content of HBsAg,HBeAg in HepG2.2.15 cells supernatant by the serum containing fresh juice’ s CN intervention and the survival rate of HepG2.2.15 cells by MTT assay.Results（1）The result showed that the CCl₄ liver-injury mice serum contents of AST,ALT and the mice liver tissue contents of MDA were declined,the ability of SOD,GSH-Px were improved,and alleviate the damage of the liver tissue by with CN;the ANIT liver-injury mice serum contents of AST,ALT,TBA,TBIL,DBILand the mice liver tissue contents of MDA were declined,and the ability of SOD,GSH-Px were improved,and alleviate the damage of the liver tissue by with CN;the serum-containing fresh juice’ s CN can improve the survival rate of L-02 cells,and decline the content of GPT,GOT,MDA,increase the ability of SOD.（2）The MTT assay showed that the serum-containing fresh juice’ s CN have a cytotoxicity effect on the HepG2.2.15 cells within a certain range of intervening time and does;the serum-containing fresh juice’ s CN were showed the inhibition of HBsAg,HBeAg for HepG2.2.15 cells,reached the peak value on the 6th day.Conclusion1.The CN have some protective effect on liver injury which induced by CC14 in vivo and in vitro.As well as,the protective mechanism is probably related with the decrease of liver anti-oxidative enzyme activities and the anti-lipid-peroxidation effect..2.The CN have improved and intervention effect on cholestasis liver injured by ANIT in mice,and that the protective effect may be carried out through normalizing functioning of the gallbladder,reducing icterus,decreasing transaminase and enhancing the immunity of the body.3.The serum-containing fresh juice’ s CN were showed the inhibition of HBsAg,HBeAg for HepG2.2.15 cells in vitro.but the mechanism needs more investigation.