摘要
目的:探讨淫羊藿次苷Ⅱ对过氧化氢诱导的人神经母细胞瘤细胞SH-SY5Y细胞氧化损伤的保护作用。方法:以过氧化氢损伤SH-SY5Y细胞建立氧化损伤模型,以不同浓度淫羊藿次苷Ⅱ(12.5、25和50μmol/L)进行干预。采用MTT法检测细胞活力,采用DCFH-DA检测ROS的水平,TUNEL法和PI染色检测细胞凋亡,免疫印迹实验检测细胞中糖原合成酶激酶3β(GSK-3β)蛋白表达。结果:不同浓度淫羊藿次苷Ⅱ(12.5、25和50μmol/L)能够抑制过氧化氢诱导的细胞死亡并呈浓度依赖性。模型组细胞氧化应激作用和凋亡率较对照组明显增强,给予淫羊藿次苷Ⅱ作用24 h后,能够显著降低活性氧(ROS)和丙二醛(MDA)含量,升高超氧化物歧化酶(SOD)活力,抑制过氧化氢诱导的细胞凋亡并上调p-ser9-GSK-3β的蛋白表达,下调p-tyr216-GSK-3β蛋白表达。结论:淫羊藿次苷Ⅱ能够抑制过氧化氢对SH-SY5Y细胞的氧化损伤,其保护作用可能与抑制氧化应激引起的细胞凋亡并调控ROS/GSK-3β信号通路有关。
Objective： To investigate protective effect of icarisideⅡagainst hydrogen peroxide-induced injury in SH-SY5 Y cells. Methods： SHSY5 Y cells were exposed hydrogen peroxide（ 200 μmol / L） were used as an in vitro model of oxidative stress. The protective effect of icarisideⅡ were detected by pretreatment with icarisideⅡ（ 12. 5,25,50 μmol / L） for 1 h prior to hydrogen peroxide challenge. Cell viability and cellular morphology were determined by MTT assay and light microscope,respectively. Reactive oxygen species（ ROS）,superoxide dismutase（ SOD） and malonaldehyde（ MDA） were detected by related kit,respectively. Cellular apotosis was detected by TUNEL assay and PI dye,respectively. Western blot was used to determined glycogen synthase kinase-3β（ GSK-3β） expression. Results： Compared with the oxidative stress case,cell viability pretreated with icarisideⅡ（ 12. 5,25,50 μmol / L） increased in a dose-dependent manner. IcarisideⅡ decreased ROS and MDA level,and increased SOD activity. Thereafter,icarisideⅡinhibited oxidative stress-induced cellular apoptosis and upregulated p-ser9-GSK-3β expression,while,downregulated p-tyr216-GSK-3β expression. Conclusion： Icariside Ⅱ has protective action against hydrogen peroxide-induced injury in SH-SY5 Y cells,which might be through apoptosis dependent ROS / GSK-3β pathway to play antioxidant effect.
作者
高健美
雷鸣
令狐浪
刘双
冯琳颖
龙隆
龚其海
Gao Jianmei Lei Ming Linghu Lang Liushuang Long Long Feng Lingying Qihai Gong(Zunyi Medical College,Zunyi 563000 Department of Pharmacology and Key Laboratory of Basic Pharmacology of Ministry of Education,Zunyi Medical University, Zunyi 563000)
出处
《中药药理与临床》
CAS
CSCD
北大核心
2016年第5期23-27,共5页
Pharmacology and Clinics of Chinese Materia Medica
基金
遵义医学院2015年度三方合作协议项目(黔科合LH字[2015]7540号)
