氧化还原调控在紫檀芪诱导HeLa细胞内质网凋亡途径中的作用

The role of redox sensing in pterostilbene-induced HeLa cell apoptosis via endoplasmic reticulum pathway

目的探讨细胞氧化还原调控在紫檀芪引起HeLa细胞凋亡过程中对内质网途径的作用。方法 SRB法评价紫檀芪对HeLa细胞的细胞毒活性。采用细胞形态学和生物化学方法检测细胞凋亡。通过荧光法对细胞内活性氧/GSH的变化进行分析。通过氧化还原相关抑制剂分析凋亡过程中的决定因素。通过半定量RT-PCR表达分析来确定内质网胁迫在紫檀芪诱导HeLa细胞凋亡中的作用。结果紫檀芪对HeLa细胞的IC50为80μmol·L-1。凋亡是紫檀芪对He-La细胞毒活性的主要表现。随着紫檀芪浓度由5μmol·L-1增加至160μmol·L-1,HeLa细胞氧化还原平衡发生明显变化。紫檀芪在80μmol·L-1～120μmol·L-1可引起内质网胁迫分子表达。抑制细胞内过氧化氢的产生会减轻紫檀芪引起的内质网胁迫并最终抑制了紫檀芪所诱导的细胞凋亡。结论细胞内氧化还原平衡的变化在紫檀芪引起HeLa细胞凋亡的内质网途径方面起了决定性的作用。

Aim To investigate the roles of intracellular redox sensing via endoplasmic reticulum pathway in pterostilbene-induced HeLa cell apoptosis.Methods The cytotoxicity was analyzed by SRB assay.The apoptosis in HeLa cells was analyzed by both cellular morphological and biochemical methods.The relative changes of the redox pair（ROS/GSH） were studied by fluorescence assay in the pterostilbene-treated HeLa cells.Redox inhibitors were also employed in the treatments to find the role of redox alteration in pterostilbene-induced HeLa cell apoptosis.Semi-quantitative RTPCR analysis was also performed to determine the role of endoplasmic reticulum stress in the pterostilbene-induced HeLa cell apoptosis.Results The IC50 of pterostilbene on HeLa cell was 80 μmol·L-1 which caused apoptosis mainly in the treatment.The cellular redox homeostasis changed with pterotilbene ranged from 5 μmol·L-1 to 160 μmol·L-1.The expression of the endoplasmic reticulum stress marks was also upregulated by the pterostilbene.However,the inhibition of intra cellular hydrogen peroxide alleviated the endoplasmic reticulum stress which finally blocked pterostilbene-induced HeLa cell apoptosis.Conclusion The imblance of cellular redox homeostasis plays a critical role in pterostilbene-induced HeLa cell apoptosis via ER-stress pathway.