NADH对L02细胞Bcl-2、Bax、P53、CD95及CD95L表达的影响

Effect of NADH on expression of Bcl-2, Bax, P53, CD95 and CD95L in L02 cells induced by ischemia-reperfusion injury

目的 研究抗氧化剂NADH对体外培养的正常人肝细胞系L0 2缺血再灌注损伤的保护作用及可能的机制。方法 实验分组 :将培养的L0 2细胞分为 :缺血再灌注损伤组 (I R) ,缺血再灌注损伤 +NADH(I R +NADH)及对照组 (未经处理的L0 2细胞 )。用流式细胞仪观察细胞处理后6、12、18及 2 4h细胞的凋亡率及 12h时 ,Bcl 2、Bax、P5 3、CD95及CD95L的表达 ,并以透射电镜观察细胞凋亡的超微结构。结果 NADH可明显抑制缺血再灌注损伤细胞的凋亡 ,并能上调Bcl 2表达 ,下调Bax、P5 3、CD95及CD95L的表达 ,与I R组相比较差异显著 (P <0 .0 5 )。透射电镜下可见典型的凋亡细胞的特征。结论 NADH对缺血再灌注损伤诱导的L0 2肝细胞有明显地保护作用。其作用机制可能与通过调节Bcl 2、Bax、P5 3。

Aim To explore a possible mechanism by which NADH protected normal human liver cell line L02 cultured in vitro from ischemia-reperfusion injury. Methods Cultured L02 cells were divided into three groups, namely, ischemia-reperfusion injury group (I/R), I/R+NADH group and control group (i.e untreated L02 cells). The apoptotic rate of L02 cells and the expressions of apoptosis-associated proteins (Bcl-2, Bax, P53, CD95 and CD95L) were analysed by flow cytometry. The ultrastructure of apoptotic L02 cells was observed under transmission electron microscope (TEM). Results NADH could inhibit distinctly apoptosis of the L02 cells induced by ischemia-reperfusion injury, up-regulated Bcl-2 and down-regulated expressions of Bax, P53, CD95 and CD95L. As compared with I/R group, these changes had statistical significance (P<0.05). Typical feature of apoptotic cells could be seen under TEM. Conclusion NADH can inhibit apoptosis of L02 cells through the medium of up-regulating the expression of Bcl-2 and down-regulating the expression of Bax, P53, CD95 and CD95L.