摘要
目的:探讨氧化应激条件下叉头框转录因子O3a(FoxO3a)参与人绒毛外滋养细胞自噬与凋亡的分子机制。方法:用葡萄糖氧化酶(GO)构建人绒毛外滋养细胞株HTR-8/SVneo的氧化应激模型并经FoxO3a特异性siRNA处理,MTT法检测细胞增殖活性,流式细胞术检测滋养细胞ROS和凋亡水平,荧光定量PCR法检测FoxO3a和BNIP3的mRNA水平,Western blot法检测FoxO3a、BNIP3和自噬相关蛋白LC3、Beclin1、p62蛋白表达水平。结果:与对照组相比,GO处理后细胞增殖活性下降,细胞内ROS含量增加,细胞内FoxO3a的mRNA和蛋白水平明显增高,FoxO3a磷酸化蛋白水平降低,自噬相关蛋白LC3II/LC3I、Beclin-1增强,p62蛋白水平降低(P<0.05),凋亡增加(P<0.05)。与阴性siRNA转染组相比,沉默FoxO3a后FoxO3a和BNIP3的mRNA和蛋白表达均明显降低,自噬相关蛋白LC3II/LC3I降低,p62蛋白表达增加(P<0.05),Beclin-1蛋白表达变化不明显(P>0.05),细胞凋亡减少(P<0.05)。结论:在GO诱导的氧化应激下,FoxO3a可通过调控BNIP3诱导滋养细胞自噬和凋亡。
Objective:To investigate the molecular mechanism of FoxO3 a involved in autophagy and apoptosis of human extravillous trophoblast cells under oxidative stress.Methods:Glucose oxidase was used to establish oxidative stress model of human extravillous trophoblastic cell line HTR-8/SVneo and treated with FoxO3 a-specific siRNA.MTT was used to detect cell proliferation activity,flow cytometry was used to detect ROS and apoptosis.Real-time PCR was used to detect FoxO3 a and BNIP3 mRNA levels.Western blot was used to detect FoxO3 a,BNIP3 and autophagy-related proteins LC3,Beclin1,p62 protein expression levels.Results:Compared with the control group,the activity of cell proliferation decreased after treatment with glucose oxidase,the intracellular ROS content increased,the mRNA and protein levels of FoxO3 a in the cells increased significantly,the protein level of p-FoxO3 a decreased,and the protein levels of LC3 II/LC3 I and Beclin1 enhanced,p62 decreased(P<0.05),the apoptosis rate of trophoblast cell increased(P<0.05).FoxO3 a and BNIP3 mRNA and protein expression were significantly reduced after silencing FoxO3 a compared with the negative siRNA transfection group,autophagy-related protein LC3 II/LC3 I decreased,p62 protein expression increased(P<0.05),Beclin-1 protein expression did not change significantly(P>0.05),the apoptosis rate of trophoblast cell decreased(P<0.05).Conclusion:FoxO3 a can induce autotrophy and apoptosis of trophoblast cell through regulation of BNIP3 under glucose oxidase-induced oxidative stress.
作者
丁燕子
张玉瑢
代延朋
邢金芳
王亮
袁恩武
Ding Yanzi;Zhang Yurong;Dai Yanpeng(Department of Clinical Laboratory,the Third Affiliated Hospital,Zhengzhou University,Zhengzhou 450052)
出处
《现代妇产科进展》
CSCD
北大核心
2019年第3期161-165,共5页
Progress in Obstetrics and Gynecology
基金
河南省科技攻关项目(No:182102310393)
