竹节参总皂苷通过JNK/Bcl-2/Bax信号通路对H2O2致SH-SY5Y神经细胞凋亡的保护作用研究

Protective Effects of Total Saponins of Panax Japonicus on H_2O_2-induced SH-SY5Y Cell Apoptosis Through JNK/Bcl-2/Bax Signaling Pathway

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 effects of total saponins of Panax Japonicus （TSPJ） on H202-inducedSH-SY5Y cell apoptosis, and to explore the underlying mechanism. Methods H202 was used to induce oxidativestress damage of SH-SYSY cells. Various concentrations of TSPJ（0.1, 1, 5, 20 μg·m L（-1）） were added into the mediumfor the pretreatment of SH-SY5Y cells for 12 hours, and then H2Oz was added for co-culturing for another 12 hours.Cell membrane integrality was identified by the release content of lactate dehydrogenase （LDH）. Intracellular reactiveoxygen species（ROS） level was measured using 2, 7-dichlorfluorescein diacetate probe under fluorescence microscope.Hoechst33342 staining was used for the detection of SH-SYSY cells, and protein expression levels of p-JNK, JNK,Bcl-2 and Bax were detected by Western blotting. Results Compared with the H202-treated group, TSPJ at 1, 5, 20μg.m-1 markedly improved the cell membrane integrality induced by H202 （P 〈 0.01 ）, and TSPJ at 0.1, 1, 5, 20μg-mL-1 reduced the rate of cell apoptosis, cleared cellular ROS （P 〈 0.01 ）, down-regulated the expression level ofp-JNK（P 〈 0.01 ） and Bax（P 〈 0.01 ）, and up-regulated protein expression of Bcl-2（P 〈 0.01 ）. Conclusion TSPJcould protect SH-SY5Y cells from H202-induced apoptosis, which probably achieved by the regulation of ROS-JNK-Bcl-2 signaling pathway.