脂多糖诱导大鼠肝细胞PARP-1依赖性细胞死亡的初步研究

Study of PARP-1 dependent cell death(PARthanatos) induced by lipopolysaccharide in rat hepatocytes

目的:观察脂多糖(lipopolysaccharide,LPS)对大鼠肝细胞聚腺苷二磷酸核糖聚合酶-1[poly(ADP-ribose)polymerase-1,PARP-1]依赖性细胞死亡的影响。方法 :大鼠肝细胞株BRL细胞常规培养,用0.1~10 mg/L LPS处理大鼠肝细胞12 h或24 h,部分实验中大鼠肝细胞LPS处理前1 h先用PARP-1抑制剂PJ-34或3-氨基苯甲酰胺(3-aminobenzamide,3-AB)预处理。用CCK-8试剂盒检测细胞活力。用蛋白免疫印迹技术检测蛋白表达。结果 :与溶剂对照组比较,LPS能剂量和时间依赖性引起大鼠肝细胞PARP-1蛋白表达上调和PARP-1活性产物多聚二磷酸核糖[poly-(ADP-ribose),PAR]修饰蛋白数量增多,大鼠肝细胞活力降低以及反映细胞凋亡的PARP-1裂解片段增多,3-AB或PJ-34部分翻转LPS诱导的大鼠肝细胞的活力降低。结论:LPS能诱导大鼠肝细胞PARP-1依赖性细胞死亡。

Objective： To elucidate the effect of lipopolysaccharide（LPS） on poly-（ADP-ribose） polymerase-1 （PARP-1） dependent cell death （PARthanatos） in rat hepatocytes. Methods： The cultured rat hepatocyte line BRL cells were exposed to 0.1-10 mg/L LPS or veehicle for 12 h or 24 h. In separate experiments, rat hepatocytes were pretreated with PARP-1 inhibitor PJ-34 or 3-AB for 1 h prior to LPS exposure. Cell viability was measured with CCK-8 kit. Proteins such as PARP- 1, poly-（ADP-ribose）（PAR）-modified proteins and apoptosis-related protein were detected with Western Blot. Results： LPS induced significant upregulations of PARP-1 and PAR-modified proteins, decreased cell viability of rat hepatocytes and enhanced cleaved 89 ku fragment of PARP-1. The decreased cell viability induced by LPS were partly reversed with PARP-1 inhibitor PJ-34 or 3-AB pretreatment. Conclusion： LPS can induce PARP-1 dependent cell death（parthanatos） in rat hepatocytes.