The Effect Of Gentianella Acuta And Scutellaria Bardadis Compatibility On The Apoptosis Of H₂₂Tumor Bearing Mice And Human Hepatocellular Carcinoma Bel-7402 Cells

ObjectiveTo investigate the effect of gentianella acuta and scutellaria bardadis compatibility on the apoptosis of H22 tumor bearing mice and human hepatocellular carcinoma Bel-7402 cells.MethodIn vitro culture:Culture human hepatocellular carcinoma cell Bel-7402 in vitro.Prepare the drug-containing serum of gentianella acuta,scutellaria barbadisd and the compatibility.The experiment were divided into blank group,gentianella acuta group,Scutellaria barbadis group,1:1 group,2:1 group,1:2 group and positive group.MTT method was used to detect the OD value of each group and calculate the cell proliferation inhibition rate.The apoptotic rate and cell cycle distribution were measured by flow cytometry.In vivo experiment:H22 tumor-bearing mice model was established and randomly divided into model group,gentianella acuta group,scutellaria barbadis group,1:1 group,2:1 group,1:2 group,positive group and blank control group.The mice were observed in the general survival state,the tumor weight was measured and the tumor inhibition rate was calculated.The tumor tissues were treated with HE staining and TUNEL staining to observe the apoptosis of tumor cells.The expression levels of Bax,Bcl-2 and caspase-3 in tumor tissues were detected by immunohistochemistry.Results1.In vitro experiment1.1 The results of MTT showed that compared with the blank group,the proliferation of Bel-7402 cells was inhibited in each group,the 2:1 group is the highest among the Chinese medicine groups.1.2 Apoptotic cells by flow cytometry results showed that comparedwith the blank group,the treatment group could induce the apoptosis of Bel-7402 cells,the apoptotic rate of 2:1 group is the highest among the Chinese medicine groups.1.3 Flow cytometry analysis showed that cell cycle distribution results:Compared with the blank group,each group could block Bel-7402 cells in G0/G1 phase and block cell proliferation,2:1 group had the most obvious cell cycle arrest among the Chinese medicine groups.2.In vivo experiments2.1 Observation of the general state of mice in each group found that the model group of mice the worst state,weight loss significantly,and rapid tumor growth.Compared with the model group,the tumor growth of each administration group was slowed down.And the weight of the traditional Chinese medicine group increased significantly,no difference compared with the blank group.2.2 The tumor weight of each group was measured and the tumor inhibition rate was calculated.It was found that each group could inhibit tumor growth,2:1 group had the highest tumor inhibition rate among the Chinese medicine groups.2.3 HE staining results showed that compared with the model group of tumor cells closely arranged,the treatment group of tumor cells arranged loose,a significant inhibitory effect.2:1 group was more effective among the Chinese medicine groups.2.4 TUNEL assay showed that the apoptotic index was increased in the visual field of each administration group compared with the model group.2:1 group was more effective among the Chinese medicine groups.2.5 Immunohistochemical test results showed that compared with the model group,each group can upregulate Bax,down-regulate Bcl-2,decrease Bax/Bcl-2,1:1 group and 2:1 group were more effective among the Chinese medicine groups.2.6 Immunohistochemical test results showed that compared with the model group,each group can upregulate caspase-3,2:1 group was more effective among the Chinese medicine groups.Conclusions1.The drug-containing serum of gentianella acuta and scuetllaria bardadis compatibility can inhibit the proliferation of human hepatocellular carcinoma cell Bel-7402,promote its apoptosis,affect cell cycle distribution,2:1 group were more effective.2.The gentianella acuta and scuetllaria bardadis compatibility can inhibits the growth of tumor cells in H22 tumor-bearing mice,induces apoptosis and improves the effect of tumor on the general state of mice,2:1 group were more effective.3.The gentianella acuta and scuetllaria bardadis compatibility can regulate Bax and Bcl-2 and activated caspase-3 to promote the role of apoptosis.