Ligustrazine-induced apoptosis is involved in its regulation of expression of Bcl-2 and Bax by caspase-dependent mechanism in rat hepatic stellate cells
Aims: Accumulating experimental and clinical studies provide clear evidence for reversibility of hepatic fibrosis (HF).Apoptosis of activated hepatic stellate cells (HSCs) has proven critical to fibrosis regression, but its mechanisms remain to be clarified.The objective of this study was to validate the apoptotic effect of lignstrazine on rat stellate cells in vitro, and elucidate the molecular mechanisms that underlie this action.Methods: Rat hepatic stellate cells were cultured in vitro.3H-TdR incorporation test and LDH release assay were used to evaluate the anti-proliferative effect of administered ligustrazine in HSCs, and flow cytometry to analyze the apoptotic ratio.Western blotting was performed to determine the expression of three key apoptotic proteins Bcl-2, Bax and caspase-3.Real-time PCR was used to detect their gene expression.Results: HSCs exposed to ligustrazine showed decreased proliferation dose-dependently, and significant apoptosis via a caspase-mediated pathway.Ligustrazine displayed anti-proliferative activity towards HSCs at 50pM and predominant pro-apoptotic effect at 70μM without evident cytotoxicity.Western blotting analysis demonstrated decreased Bcl-2 expression and growing expression of Bax and caspase-3.Real-time PCR experiments indicated that ligustrazine could correspondingly regulate the gene expression of Bcl-2, Bax and caspase-3.Ligustrazine was able to induce HSC apoptosis in rats in vitro depending on a cytochondriome-mediated pathway involving Bcl-2, Bax and caspase-3.Conclusion: This investigation demonstrated the promising role of ligustrazine that may contribute to hepatic fibrosis regression through stimulation of HSC apoptosis.This apoptotic effect was modulated by cytochondriome pathway involving down-regulation of Bcl-2 and up-regulation of Bax and caspase-3 at both gene and protein levels.Elucidation of the underlying mechanisms helped to provide new evidence for drug-facilitated HSC apoptosis.Ligustrazine may be exploited as a potential option for treating and reversing hepatic fibrosis.
Hepatic fibrosis hepatic stellate cells fibrosis reversal ligustrazine apoptosis caspase
Feng Zhang Na Lei Jin Ma Yin Lu Shizhong Zheng
Department of Clinical Pharmacy, Nanjing University of Chinese Medicine, Nanjing, 210029, China
国内会议
江苏省药理学会青年工作委员会成立大会暨药理学科青年科技创新学术研讨会
苏州
英文
172-173
2013-05-24（万方平台首次上网日期，不代表论文的发表时间）