Inhibition Effects Of Allicin And Flos Lonicerae On The Cells Apoptosis Induced By VSV And The Possible Molecular Mechanism Involved

Objective To explore the inhibition effects of Chinese herbs allicin and flos lonicerae on VSV -induced apoptosis in the THP-1,MT-2 and WISH Cells and their possible molecular mechanism involved.Methods First confirm the safe concentrations of allicin and flos lonicerae by MTT assay,then select three appropriate concentrations of the two herbs as the experimental concentrations. Make three groups of the THP-1, MT-2 and WISH cells respectively as normal cells group, VSV-only deal cells group and allicin/ flos lonicerae+VSV acting cells group. Pre-incubate the THP-1,MT-2 and WISH cells with different concentrations (50% 25 and 10μg/ml of allicin; 10 mg/ml, 5 mg/ml and 1 mg/mlof flos lonicerae) of the two herbs for 4 h,then put the VSV into the cell culture with MOI=10:1(10 virus infect 1 cell) VSV dose for another 72 h.Based on the model of apoptosis of THP-1, MT-2 and WISH cells induced by VSV in vitro,cell morphological observation was carried out with the inverted electron microscope. Cell viability was tested by MTT assay. Flow cytometry by Annex in V/PI double stain was performed to quantify the apoptotic cells. The expressions of Caspase-3,6, 7, 9 and NF-κ B proteins were tested by Western blot.Results Cell morphological observation with the inverted electron microscoperesulted in significant changes of cell apoptosis treated by VSV only and oubvious conversion of the VSV-induced cells group trated by allicin and flos lonicerae. We could see ftom MTT assay that different concentrations of allicin and flos lonicerae can increase the cell viability respectively,and the results have statistical significance. Flow cytometry assay indicated that VSV can markedly increase the apoptosis rate of the THP-1, MT-2 and WISH cells. but after treated with allicin and flos lonicerae,the apoptosis rate decreased in different degrees,especially the high concentrations of the two herbs,suggesting a dose-dependent manner. Western blot detection showed that VSV could activate the expression of the Caspase-3,6, 7, 9 and NF-κ B proteins evidently,and when they were treated by different concentrations of allicin and flos lonicerae,the expression of the proteins gradually decreased following the different concentrations of the two herbs,and the most inhibiting effection of the high concentrations of the two herbs were observed.Conclusion Allicin and flos lonicerae could prevent VSV-induced THP-1,MT-2 and WISH cells from apoptosis in different degrees,of which the inhibition of the expression of NF-κB and caspase 3 ,6 ,7 ,9 are observed and considered as the possible signalling pathway of anti-apoptosis, this may also be the potential anti-virus mechansim of allicin.and flos lonicerae.