HtrA2/Omi差异化调控年轻细胞及衰老细胞的应激敏感性  

HtrA2/Omi Differentially Regulates the Stress Sensitivity of Young and Senescent Cells

作　　者：杨婵 太颢然 李小双 苏炳银[1,3] 李淑蓉[1,2] Yang Chan;Tai Haoran;Li Xiaoshuang;Su Bingyin;Li Shurong(Key Laboratory of Development and Regeneration of Sichuan Province, Chengdu Medical College,Chengdu 610500,China;Department of Pathology and Pathophysiology, Chengdu Medical College,Chengdu 610500, China;Department of Anatomy and Histology and Embryology, Chengdu Medical College, Chengdu 610500, China)

机构地区：[1]成都医学院发育与再生重点实验室,成都610500 [2]成都医学院病理学与病理生理学教研室,成都610500 [3]成都医学院人体解剖与组织胚胎学教研室,成都610500

出　　处：《成都医学院学报》2020年第3期307-313,共7页Journal of Chengdu Medical College

基　　金：国家自然科学基金(No:31801013);四川省科技厅应用基础研究重点项目(No:2019YJ0366,No:2017JY0138);发育与再生四川省重点实验室研究基金项目(No:SYS17-002)。

摘　　要：目的探讨HtrA2/Omi对年轻细胞和衰老细胞应激下凋亡发生的作用。方法建立氧化应激诱导的人脐静脉内皮细胞(HUVECs)衰老模型;构建可诱导过表达野生型(WT)和酶活性失活突变型HtrA2(S306A)/Omi的慢病毒Tet-On系统,用HUVECs建立稳转株;利用不同浓度的H2O2处理上述细胞株;衰老相关β半乳糖苷酶(SA-β-gal)染色检测细胞衰老,TUNEL检测细胞凋亡,蛋白免疫印迹及qRT-PCR检测HtrA2蛋白及mRNA水平。结果利用100μmol/L H2O2处理细胞1 h成功建立氧化应激诱导的衰老模型;衰老细胞中HtrA2/Omi的蛋白及mRNA水平相对于年轻细胞升高;年轻细胞受氧化应激后,过表达野生型HtrA2/Omi的细胞凋亡数量少于对照组和过表达突变型HtrA2/Omi组;而衰老细胞受氧化应激后,过表达野生型HtrA2/Omi的细胞发生凋亡的数量多于对照组和过表达突变型HtrA2/Omi组。结论年轻细胞中过表达HtrA2/Omi能够抑制凋亡,而在衰老细胞中过表达HtrA2/Omi则会诱导凋亡,因此HtrA2/Omi能够增强年轻细胞对氧化应激的抗性,在衰老细胞中则发挥相反的作用。Objective To explore the effect of HtrA2/Omi on the apoptosis of young cells and senescent cells under stress.Methods Establish oxidative stress-induced aging model of human umbilical vein endothelial cells(HUVECs).Construct a lentivirus Tet-On system that can induce overexpression of wild-type(WT)HtrA2/Omi and enzymatically inactive mutant(S306A)HtrA2/Omi,and establish stable transfectants with human umbilicalvascular endothelial cells(HUVECs).The above cell lines were treated with different concentrations of H2O2.Senescence-associatedβ-galactosidase(SA-β-gal)staining was used to detect cell senescence,TUNEL was used to detect apoptosis,western blot(WB)was used to detect protein levels,and qRT-PCR was used to detect mRNA levels.Results An oxidative stress-induced aging model was successfully established by treating cells with 100μmol/L H2O2 for 1 h.The protein and mRNA levels of HtrA2/Omi in senescent cells were increased relative to those in young cells.When young cells were subjected to oxidative stress,overexpressed wild-type HtrA2/Omi cells had less apoptosis than control cells and overexpressed mutant HtrA2/Omi cells.However,when senescent cells were subjected to oxidative stress,overexpressing wild-type HtrA2/Omi cells underwent more apoptosis than control cells and overexpressed mutant HtrA2/Omi cells.Conclusion Overexpression of HtrA2/Omi in young cells can inhibit apoptosis,while overexpression of HtrA2/Omi in senescent cells can induce apoptosis.Therefore,HtrA2/Omi can enhance the resistance of young cells to oxidative stress,but plays the opposite role in senescent cells.

关 键 词：HtrA2/Omi 氧化应激 细胞衰老 细胞凋亡 

分 类 号：Q291[生物学—细胞生物学]