The Effect Of Mangiferin On Proliferation,Apoptosis,IL-4 Secretion Of BCBL-1 Cell And KSHV Replication Cycle

Objective: The aim of this study was to investigate the effects of mangiferin on the proliferation,apoptosis,secretion of IL-4 and intracellular KSHV in BCBL-1 cell in vitro and discuss its possible mechanism.Methods: 1.CCK-8 assay was used to detect the inhibitory effect of different concentrations of Mangiferin on BCBL-1 cells in vitro and to calculate the half inhibitory concentration(IC50).2.Using the Enzyme Linked Immunosorbent Assay(ELISA)to detect the secretion of IL-4 in supernatant of BCBL-1 cells treated with 800 μmol/L of mangiferin for 12 h,24h,48 h.3.To analyze the apotosis,BCBL-1 cells,which were treated with mangiferin(400,800 or 1600 μmol/L)for 24 h and 48 h,were labeled with Annexin V/PI-FITC and analysed by flow cytometry analysis.4.The expression of the KSHV replication cycle related genes(LANA,ORF50 and PAN)was detected by the real-time fluorescent quantitative PCR assay following that BCBL-1 cells were treated with 400,800 and 1600 μmol/L of mangiferin for 24 h,48h and 72 h.5.The variant expression of the apoptosis related signal pathway gene was investigated by gene chip screening with RT2 Profiler PCR Arrays-Human Apoptosis after 800 μmol/L mangiferin intervention on BCBL-1 cells for 12 h.Results: 1.The proliferation of BCBL-1 cells was inhibited by mangiferin,which was dose-dependent(P < 0.05).The IC50 of mangiferin to BCBL-1 cells are 979.89 μmol/L(24h)and 739.19 μmol/L(48h)respectively.2.The expression of IL-4 was increased in the supernatant of 800 μmol/L mangiferin treated BCBL-1 cells,and the difference was statistically significant(P < 0.01).3.The cell apoptosis assay of Annexin V/PI-FITC labeled showed that mangiferin had no obvious effect on inducing the apotosis of BCBL-1 cells(P > 0.05).4.There was no significant change in the expression of the latent gene LANA after treatment with mangiferin,but the expression of ORF50 and PAN of the viral lytic phase were increased(P <0.05).5.The expression of the apoptosis related genes,CASP5(-2.32,P = 0.049),FASLG(-2.69,P = 0.040),were decreased by using RT2 Profiler PCR Arrays-Human Apoptosis.Conclusion: 1.Mangiferin inhibit the proliferation of BCBL-1 cells,and its mechanism may be related to the secretion of IL-4.2.Mangiferin could induce BCBL-1 cell to death or necrosis,but the effect on BCBL-1 cells apoptosis was negligible,which may be related to the interference of virus that led to the downregulation in the transcription levels of apoptosis related genes CASP5 and FASLG in BCBL-1 cells.3.The treatment of mangiferin can promote the virus KSHV into the lytic phase in BCBL-1 cells.