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目的探讨竹节参总皂苷对过氧化氢(H_2O_2)引起的人神经瘤母瘤细胞(SH-SY5Y)凋亡的保护作用及分子机制。方法用H_2O_2作用SH-SY5Y细胞建立凋亡模型。以不同浓度竹节参总皂苷(0.1,1,5,20μg·m L~(-1))预处理12 h,再加入H_2O_2继续培养12 h。通过乳酸脱氢酶(Lactate dehydrogenase,LDH)释放量检测细胞膜完整性;DCFH-DA探针法检测活性氧(Reactive Oxygen species,ROS)含量;Hoechst33342染色法检测细胞凋亡;Western blot法检测p-JNK,JNK,Bcl-2,Bax蛋白表达水平。结果竹节参总皂苷(1,5,20μg·m L~(-1))能显著改善细胞膜完整性(P<0.01);竹节参总皂苷(0.1,1,5,20μg·m L~(-1))能降低细胞凋亡程度,清除细胞内ROS(P<0.01);并且下调p-JNK及Bax的表达,上调Bcl-2蛋白的表达(均P<0.01)。结论竹节参总皂苷对H_2O_2所致的SH-SY5Y细胞凋亡具有保护作用,其可能是通过调控ROS-JNK-Bcl-2通路发挥作用。
Objective To investigate the protective effect of SSP on the apoptosis of human neuroblastoma (SH-SY5Y) induced by hydrogen peroxide (H_2O_2) and its molecular mechanism. Methods H_2O_2-induced SH-SY5Y cell apoptosis model. The cells were pretreated with 0.1, 1, 5 and 20 μg · m L -1 of SSP for 12 h and then cultured with H 2 O 2 for 12 h. The integrity of cell membrane was detected by the release of lactate dehydrogenase (LDH); the content of reactive oxygen species (ROS) was detected by DCFH-DA probe; the apoptosis was detected by Hoechst33342 staining; the expression of p- JNK, JNK, Bcl-2, Bax protein expression levels. Results SSP (1,5,20 μg · m L -1) could significantly improve cell membrane integrity (P <0.01). Total ginsenosides (0.1,1,5,20 μg · m L ~ (-1)) could reduce the degree of apoptosis, clear the intracellular ROS (P <0.01), and down-regulate the expressions of p-JNK and Bax, and up-regulate the expression of Bcl-2 protein (all P <0.01). Conclusion SSP can protect SH-SY5Y cells induced by H_2O_2, which may play a role in the regulation of ROS-JNK-Bcl-2 pathway.