紫铆因通过抑制p53信号通路对抗丙酮醛诱导的PC12细胞凋亡

Butein protects from PC12 cells apoptosis induced by methylglyoxal via inhibiting p53 signaling pathway

目的探讨紫铆因对丙酮醛诱导的PC12细胞凋亡的影响及作用机制。方法采用不同剂量的紫铆因预处理1 h后,1.5 mmol·L^(-1)丙酮醛诱导PC12细胞凋亡,MTT及LDH比色法分析细胞存活率及细胞毒性;PI及Hoechst 33342双染法分析细胞凋亡及坏死;逆转录PCR检测促凋亡基因p53、caspase-9和抗氧化基因SOD2的表达;免疫印迹法检测p53、caspase-9的蛋白表达。结果不同剂量的紫铆因预处理PC12细胞1 h后,可明显对抗丙酮醛引起的细胞凋亡,提高细胞存活率,减少细胞核固缩、碎裂,抑制促凋亡基因p53、caspase-9的过表达,提高抗氧化基因SOD2的表达。结论紫铆因能明显对抗丙酮醛诱导的PC12细胞凋亡,作用机制与其抗氧化活性,降低促凋亡基因p53、caspase-9的表达有关。

Aim To study the effect of butein on apoptosis of PC12 cells induced by methylglyoxal(MG)and its mechanism.Methods Being pretreated with different concentrations of butein,PC12 cells were damaged by 1.5 mmol·L-1 MG.Cell viability and cell toxicity were evaluated by MTT and LDH assay.Cell apoptosis and death were analyzed by PI and Hoechst 33342.The antioxidant gene and proapoptotic gene expressions were determined by RT-PCR.The protein expression of p53 was detected by Western blot.Results Being pretreated with 2.5~10μmol·L-1 butein for 1 h significantly increased the cell viability,decreased LDH release,and protected from cell nuclei shrinkage,condensation and cleavage by MG.Meanwhile,butein increased the gene expression of SOD2,decreased the gene expression of proapoptotic genes p53 and caspase-9,and lowered the protein expression of p53.Conclusion Butein can protect apoptosis of PC12 cells from MG in a dose-dependent manner,which is linked with antioxidation and inhibiting p53 and caspase-9 gene expression.