摘要
目的探讨黄杞苷对H_(2)O_(2)诱导SH-SY5Y细胞损伤的保护作用与分子机制。方法黄杞苷(5、10、20、40、80和160μmol·L-1)预处理SH-SY5Y细胞12 h,MTT法检测黄杞苷对H_(2)O_(2)诱导损伤后细胞活力的影响;通过氮蓝四唑(NBT)法总超氧化物歧化酶(SOD)活力检测试剂盒和总谷胱甘肽过氧化物酶(NADPH)检测试剂盒测定黄杞苷对H_(2)O_(2)诱导损伤后SOD和谷胱甘肽过氧化物酶(GSH-Px)活力的影响;通过流式细胞术对SH-SY5Y细胞进行活性氧(ROS)水平的测定。黄杞苷(20μmol·L^(-1))预处理SH-SY5Y细胞12 h,H2O2孵育6、12和24 h或用黄杞苷(5、10和20μmol·L^(-1))预处理细胞12 h,H2O2孵育12 h,采用Western blot法分别检测Nrf2及HO-1表达情况。结果与模型组比较,黄杞苷能够提高SH-SY5Y细胞的活力,显著提高SOD和GSH水平,减少ROS产生(P<0.05)。黄杞苷能够促进Nrf2蛋白由细胞质向细胞核内转移,黄杞苷(20μmol·L^(-1))预处理SH-SY5Y细胞12 h时转移达到顶峰。H_(2)O_(2)损伤模型下,黄杞苷浓度依赖性地促Nrf2的核内转移(P<0.05),并促进下游蛋白HO-1的表达(P<0.05)。结论黄杞苷对H2O2诱导SH-SY5Y细胞损伤具有保护作用,其作用机制与抗氧化,促进Nrf2、HO-1的表达有关。
Objective To investigate the protective effect and molecular mechanism of engeletin on SH-SY5Y cells injured by H_(2)O_(2).Methods SH-SY5Y cells were pretreated with engeletin(5,10,20,40,80 and 160μmol·L^(-1))for 12 h.MTT assay was used to detect the effect of engeletin on cell viability after H_(2)O_(2) induced injury.The effects of engeletin on the activities of superoxide dismutase(SOD)and glutathione peroxidase(GSH PX)after H_(2)O_(2) induced injury were determined by NBT method and NADPH method;the levels of reactive oxygen species(ROS)in SH-SY5Y cells were determined by flow cytometry.SH-SY5Y cells were pretreated with engeletin(20μmol·L^(-1))for 12 h and incubated with H_(2)O_(2) for 6 h,12 h and 24 h,or pretreated with engeletin(5,10 and 20μmol·L-1)for 12 h and incubated with H_(2)O_(2) for 12 h.The expressions of Nrf2 and HO-1 were detected by Western blot.Results Engeletin could protect SH-SY5Y cells against H2O2 induced injury.Compared with the model group,engeletin could increase the cell viability,significantly increase the levels of SOD and GSH,and reduce the production of ROS(P<0.05).Engeletin promoted the transfer of Nrf2 protein from cytoplasm to nucleus,and reached the peak when SH-SY5Y cells were pretreated with engeletin(20μmol·L^(-1))for 12 h.In the model group of H_(2)O_(2) induced injury,engeletin promoted the nuclear transfer of Nrf2 in a concentration dependent manner(P<0.05),and promoted the expression of HO-1,a downstream protein of Nrf2(P<0.05).Conclusion Engeletin protected SH-SY5Y cells from H_(2)O_(2) induced injury,and its mechanism was related to antioxidation and promoting the expression of Nrf2 and HO-1.
作者
刘慧
闫冬
徐洋洋
李淑翠
张树平
LIU Hui;YAN Dong;XU Yangyang;LI Shucui;ZHANG Shuping(Department of Pharmacology,Binzhou Medical University,Yantai 264003,Shandong,P.R.China;Department of Pharmacy,Binzhou Medical University Hospital,Binzhou 256603,Shandong,P.R.China)
出处
《滨州医学院学报》
2021年第1期1-6,共6页
Journal of Binzhou Medical University
基金
国家自然科学基金(31570352,31170321)。
