人参皂苷F2干预Caspase级联反应抑制H2O2诱导的细胞凋亡

Ginsenoside F2 Interferes with the Cascade Reactions of Caspase to Inhibit H2O2-induced Apoptosis

摘要:

本研究探讨了人参皂苷F2对H2O2诱导人胚肾HEK-293细胞损伤的抗凋亡作用。利用试剂盒测定凋亡细胞Caspase-6和Caspase-9活性水平，应用Western Blot检测凋亡细胞中Bcl-2凋亡家族（Bcl-2和Bax）与Caspase凋亡家族（Cleaved Caspase-3和Cleaved PARP）的蛋白表达量。H2O2损伤细胞后，Caspase酶活力提高，促凋亡蛋白表达提高；1.25、5、20 μmol/L F2预处理损伤细胞后，Caspase-6和Caspase-9活力呈浓度依赖性降低（p<0.05），当F2浓度达到20 μmol/L时，Caspase-6活性降低了6.83 U/mg protein，Caspase-9活性降低了5.88 U/mg protein；Cleaved Caspase-3和Cleaved PARP蛋白表达量显著低于损伤组（p<0.05），随着F2浓度的升高，Cleaved Caspase-3表达量分别下降至280.90%、232.46%、185.26%，Cleaved PARP表达量随着F2浓度升高而降低至110.36%、85.85%、61.95%；F2高浓度组的Bcl-2/Bax比值（73.94%）较损伤组比显著提升（p<0.05）。表明人参皂苷F2可以抑制凋亡蛋白Caspase的活性，降低Bax促凋亡蛋白与Caspase家族上、中、下游蛋白的表达，通过调控Caspase级联反应抑制H2O2刺激引起的细胞凋亡。

Abstract:

In this study, the anti-apoptotic effect of ginsenoside F2 on H2O2-induced injury in human embryonic kidney HEK-293 cells was evaluated. Using commercial kits, the activities of Caspase-6 and Caspase-9 of apoptotic cells were examined by measured. Western blot tests were conducted to detect the protein expression of the Bcl-2 family (Bcl-2 and Bax) and Caspase family (Cleaved Caspase-3 and Cleaved PARP). After the cells were injured by H2O2, the activity of Caspase increased and the expressions of pro-apoptotic proteins increased. The activities of Caspase-6 and Caspase-9 decreased in a concentration-dependent manner (p<0.05) after the pretreatment of cells with 1.25, 5, 20 μmol/L ginsenoside F2 to injure cells (p<0.05). At the F2 concentration of 20 μmol/L, the activities of Caspase-6 and Caspase-9 decreased by 6.83 U/mg protein and 5.88 U/mg protein., respectively, and the protein expression levels of Cleaved Caspase-3 and Cleaved PARP were significantly lower than those of the injured group (p<0.05). With the increase of F2 concentration, the expression of Cleaved Caspase 3 dropped to 280.90%, 232.46% and 185.26%, respectively. The expressions of Cleaved PARP decreased to 110.36%, 85.85% and 61.95%, respectively, with the increase of F2 concentration. The ratio of Bcl-2/Bax for the high F2 concentration group (73.94%) was significantly higher than that of the injured group (p<0.05). These results showed that ginsenoside F2 can inhibit the activity of apoptotic protein Caspase, and reduce the expression of Bax pro-apoptotic protein and the upstream, midstream and downstream proteins of the Caspase family, through inhibiting H2O2-induced apoptosis via regulating the Caspase cascade reactions.