摘要
目的:研究匹诺塞林对缺糖缺氧/复糖复氧(OGD/R)诱导SH-SY5Y细胞损伤的保护作用。方法:SH-SY5Y细胞系制备OGD/R损伤模型,缺糖缺氧2 h,复糖复氧24 h。MTT法检测细胞存活率,通过LDH释放率考察细胞活力,JC-1荧光探针检测线粒体膜电位,应用DCFH-DA和MitoSOX分别检测细胞ROS及线粒体超氧化物水平,Western Blot法测定cytochrome c,Bcl-2,Bax,cleaved caspase 3蛋白表达水平,免疫荧光法测定HO-1和Nrf2的表达。结果:OGD/R导致SH-SY5Y细胞存活率显著降低。匹诺塞林(1和10μmol·L-1)能显著增加SH-SY5Y细胞存活率,降低细胞凋亡率。同时,匹诺塞林能够减少OGD/R诱导ROS的释放,减缓线粒体膜电位去极化,促进HO-1蛋白表达及Nrf2核转位,降低OGD/R损伤所致cytochrome c,Bax,cleaved caspase 3的升高,抑制Bcl-2的降低。结论:匹诺塞林能够抑制OGD/R损伤引起的SH-SY5Y细胞氧化应激与线粒体凋亡。
Objective:To study the protective effect of pinocembrin on SH-SY5Y cells subjected to oxygen glucose deprivation/reperfusion (OGD/R)injury.Methods:The OGD/R model was established via 2h of oxygen/glucose deprivation followed by 24h of reperfusion in SH-SY5Y cells.The cell viability was measured by MTT assay,and investigated through the release rate of LDH.MitochondriaI membrane potential was detected by JC-1fluorescence probe.Cellular ROS levels and mitochondria superoxide levels were detected by the DCFH-DA and MitoSOX probes,respectively.Protein expression levels of cytochrome c,Bcl-2,Bax,and cleaved caspase 3 were detected by Western blot method.And the expression of HO-1and the nuclear translocation of Nrf2were determined by immunofluorescence assay.Results:OGD/R resulted in significant decrease of cell viability,and pinocembrin could significantly increase the viability of SH-SY5Y cells.Meanwhile,pinocembrin could reduce the release of ROS induced by OGD/R,and slow down the mitochondrial membrane potential depolarization.Further study showed that it promoted HO-1expression and Nrf2nuclear translocation,reduced the increase of cytochrome c,Bax,cleaved caspase 3,and inhibited the decrease of Bcl-2.Conclusion:Pinocembrin can inhibit oxidative stress and cell apoptosis via ROS-mitochondria pathway in SH-SY5Y cells damaged by OGD/R.
作者
张雯
宋俊科
周启蒙
王海港
梁宇
杜冠华
ZHANG Wen;SONG Jun-ke;ZHOU Qi-meng;WANG Hai-gang;LIANG Yu;DU Guan-hua(Beijing Key Laboratory of Drug Target Identification and Drug Screening,Institute of Materia Medica,Chinese Academy of Medical Sciences &Peking Union Medical College,Beijing 100050,China)
出处
《中国新药杂志》
CAS
CSCD
北大核心
2018年第22期2651-2658,共8页
Chinese Journal of New Drugs
基金
中国医学科学院医学与健康科技创新工程资助项目(2017-12M-1-010)
国家自然科学基金资助项目(81603100)
抗毒药物与毒理学国家重点实验室开放课题资助项目(TMC201510)
北京协和医学院研究生创新基金资助项目(2015-1007-07)
