芝麻素对高糖损伤SH-SY5Y细胞的保护效果及机制

Protective effect of sesamin on high glucose-induced neurotoxicity in SHSY5Y cells

目的研究芝麻素(sesamin)对高糖所致人神经母细胞瘤(SH-SY5Y)细胞损伤的保护作用及其机制。方法SH-SY5Y细胞接种于含100 mmol/L葡萄糖的培养基并与10、20、40μmol/L芝麻素共同孵育36 h,MTT法测定细胞存活率,比色法测定细胞培养液乳酸脱氢酶(LDH)水平,ELISA检测DNA片段及半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)活性。2',7'-二氯荧光黄双乙酸盐(DCFH-DA)探针法检测细胞中活性氧(ROS)的水平,试剂盒检测细胞内丙二醛(MDA)、超氧化物岐化酶(SOD)、过氧化氢酶(CAT)和还原型谷胱甘肽(GSH)活性。光度法检测细胞烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶活性,实时荧光定量检测NADPH氧化酶亚基p47phox mRNA表达。Western blot法检测p47phox蛋白表达。结果芝麻素可增加高糖作用下SH-SY5Y细胞的存活率,抑制细胞LDH释放,阻止高糖诱导的DNA断裂,降低Caspase-3活性。同时芝麻素能够增加高糖作用下SH-SY5Y细胞内SOD、CAT和GSH的活性,清除细胞内的ROS,抑制NADPH氧化酶活性及p47phox mRNA和蛋白的表达。结论芝麻素对高糖诱导SY5Y细胞损伤具有保护作用,其机制可能与抗氧化和抑制NADPH氧化酶活性有关。

AIM To explore the effects of sesamin on cell proliferation and its protective role in cellular oxida- tire injury induced by high glucose to SH-SY5 cell line. METHODS SH-SY5Y cell in 100 mmol/L glucose medium was incubated with 10, 20, 40 μmol/L sesamin for 36 h. MTF assay was used to evaluate the cell prolif- eration of SH-SY5Y cells. In the meantime, the levels of lactate dehydrogenase （LDH）, NADPH oxidase, super- oxide dismutase （SOD）, catalase （CAT） as well as the levels of malondialdehyde （MDA）, glutathione （GSH）, intracellular reactive oxygen species （ROS）, Caspase-3 activity and DNA fragmentation were measured with or without sesamin treatment. RESULTS The treatment of SH-SY5Y cells with sesamin was associated with a marked diminution in Caspase-3 expression, LDH level, DNA fragmentation, intracellular ROS and MDA levels. Additionally, the cell viability, GSH level, SOD and CAT activity were notably elevated by sesamin, whereas the activity of NADPH oxidase, the mRNA and the protein expression of NADPH oxidase subunit p47phox were attenu- ated. CONCLUSION Sesamin can attenuate the high glucose-induced neurotoxicity in SH-SYSY cells. Its pro- tective effects may be related to its antioxidant action and the inhibition of NADPH oxidase.