Experimental Study On Apoptosis And Autophagy Of HepG2 Cells Induced By Serum Containing Xiaochaihu Decoction

Purpose：To investigate the interrelation of autophagy and apoptosis in theanticancer effects of Xiao-Chaihu Decoction(XCHD)on HepG2cells.Materials and methods：In this study, a serum pharmacological method wasemployed to investigate the interrelation of autophagy and apoptosis. HepG2cells were treated with various concentrations of XCHD at different timeintervals, and the inhibitory effect was evaluated using MTT and morphologicalobservation. The proapoptotic effects in different time were determined by thechange of mitochondrial membrane potential(Rh123staining)and nuclearmorphology(hoechst33342staining). The levels of apoptosis-related proteinexpression(cytochrome c), evaluated by western blot analysis. To furtherinvestigation the interaction of autophagy and apoptosis in anticancer effectsof XCHD on HepG2cells, the apoptosis was blocked by selectiveinhibitor(caspase-3inhibitor). The effect to autophagy were detected by theformation of autophagic vacuoles(MDC staining), and the expression ofautophagy-related proteins(Beclin-1, LC3).Results：1.XCHD inhibited the growth of HepG2cells.2.XCHD treatment significantly caused reduction of mitochondrial membranepotential and condensation of nuclear chromatin. The results of western blotanalysis showed that the expression of pro-apoptotic protein(cytochrome c)wereincreased.3.The detection of XCHD-induced autophagy shown that XCHD treatment resultedin accumulation of MDC-labeled vesicles in HepG2cells, and time-dependentlyupregulated Beclin-1expression, enhanced LC3-I to LC3-II conversion in HepG2cells. While the inhibitor weaken XCHD-induced cell autophagy in HepG2cellsto the treatment group. Conclusions：1.This study suggests that apoptosis has participated XCHD-induced cell deathin HepG2cells from started to finished.2.Apoptosis promoted autophagy in the process of XCHD inhibited the growth ofHepG2cells.