CircACTR2调节miR-345-3p/TXNRD1轴对高糖诱导的滋养层细胞增殖、凋亡的影响  

Impacts of CircACTR2 on the proliferation and apoptosis of trophoblast cells induced by high glucose by regulating miR-345-3p/TXNRDI axis

作　　者：罗亚丽[1] 黄志刚[2] 罗思通 徐洲 LUO Yali;HUANG Zhigang;LUO Sitong;XU Zhou(Department of Obstetrics and Gynecology,Bazhou Qu Fu You Bao Jian Yuan,Bazhong,Sichuan 636000,China;Department of Experimental Medicine,West China Hospital of Sichuan University,Chengdu,Sichuan 610041,China;Sichuan Jinxin Women's and Children's Hospital,Chengdu,Sichuan 610066,China)

机构地区：[1]巴中市巴州区妇幼保健院妇产科,四川巴中636000 [2]四川大学华西医院实验医学科,四川成都610041 [3]四川锦欣妇女儿童医院妇产科,四川成都610066

出　　处：《中国优生与遗传杂志》2023年第5期957-962,共6页Chinese Journal of Birth Health & Heredity

基　　金：2022年成都市医学科研课题立项项目(2022586)。

摘　　要：目的 探究CircACTR2对高糖诱导的滋养层细胞增殖、凋亡的影响,探讨其作用机制。方法 将人绒毛膜滋养层细胞(HTR-8/Svneo)分为:control组(5 mmol/L葡萄糖)、高糖组(25 mmol/L葡萄糖)、si-NC组(转染si-NC)、si-CircACTR2组(转染si-CircACTR2)、si-CircACTR2+inhibitor-NC组(si-CircACTR2和inhibitor-NC共转染)、si-CircACTR2+miR-345-3p inhibitor组(si-CircACTR2和miR-345-3p inhibitor共转染)。RT-qPCR检测细胞中CircACTR2、miR-345-3p的表达;MTT法检测细胞增殖;流式细胞仪检测细胞凋亡;Westernblot检测细胞中硫氧还蛋白还原酶1(TXNRD1)、增殖细胞核抗原(PCNA)、Bcl-2相关X蛋白(Bax)、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(cleaved cleaved caspase-3)的表达;双荧光素酶报告基因实验分别验证CircACTR2和miR-345-3p、miR-345-3p和TXNRD1的关系。结果 与control组相比,高糖组HTR-8/Svneo细胞的凋亡率、CircACTR2、TXNRD1、Bax、cleavedcaspase-3蛋白表达升高(P<0.05),miR-345-3p、OD490值、PCNA蛋白表达降低(P<0.05);与高糖组、si-NC组相比,si-CircACTR2组HTR-8/Svneo细胞的凋亡率、CircACTR2、TXNRD1、Bax、cleaved caspase-3蛋白表达降低(P<0.05),miR-345-3p、OD490值、PCNA蛋白表达升高(P<0.05);与si-CircACTR2组、si-CircACTR2+inhibitor-NC组相比,si-CircACTR2+miR-345-3p inhibitor组HTR-8/Svneo细胞的miR-345-3p、OD490值、PCNA蛋白表达降低(P<0.05),凋亡率、TXNRD1、Bax、cleaved caspase-3蛋白表达升高(P<0.05);CircACTR2靶向调控miR-345-3p表达,miR-345-3p靶向负调控TXNRD1表达。结论 敲低CircACTR2可能通过靶向miR-345-3p来下调TXNRD1表达,进而抑制高糖诱导的滋养层细胞凋亡,促进滋养层细胞增殖。Objective To explore the effect of CircACTR2 on the proliferation and apoptosis of trophoblast cells induced by high glucose,and to investigate its mechanism.Methods Human chorionic trophoblast cells(HTR-8/Svneo)were separated into:Control group(5 mmol/L glucose),high glucose group(25 mmol/L glucose),si-NC group(transfection of si-NC),si-CircACTR2 group(transfection of si-CircACTR2),si-CircACTR2+inhibitor-NC group(co-transfection of si-CircACTR2 and inhibitor-NC),si-CircACTR2+miR-345-3p inhibitor group(co-transfection of si-CircACTR2 and miR-345-3p inhibitor).RT-qPCR was applied to detect the expression of CircACTR2 and miR-345-3p in cells.MTT assay was applied to detect cell proliferation;flow cytometry was applied to detect apoptosis of cells.Western blot was applied to detect the expression of thioredoxin reductase 1(TXNRD1),proliferating cell nuclear antigen(PCNA),Bcl-2-associated X protein(Bax),and activated cysteine-containing aspartate proteolytic enzyme 3(cleaved caspase-3)in cells.Dual-luciferase reporter assay was applied to verify the relationship between CircACTR2 and miR-345-3p,miR-345-3p and TXNRD1,respectively.Results Compared with the control group,the apoptosis rate,the protein expressions of CircACTR2,TXNRD1,Bax,and cleaved caspase-3 of HTR-8/Svneo cells in the high glucose group increased(P<0.05),the expression of miR-345-3p,OD490,and the expression of PCNA protein decreased(P<0.05),compared with the high glucose group and the si-NC group,the apoptosis rate of HTR-8/Svneo cells and the protein expressions of CircACTR2,TXNRD1,Bax and cleaved caspase-3 in the si-CircACTR2 group decreased(P<0.05),the expression of miR-345-3p,OD490,and the expression of PCNA protein increased(P<0.05),compared with si-CircACTR2 group and si-CircACTR2+inhibitor-NC group,the expression of miR-345-3p,OD49o value,and PCNA protein expression of HTR-8/Svneo cells in si-CircACTR2+miR-345-3p inhibitor group decreased(P<0.05),the apoptosis rate,the protein expression of TXNRD1,Bax and cleaved caspase-3 increased(P<0.05).Circ

关 键 词：CircACTR2 miR-345-3p/TXNRD1轴 滋养层细胞 增殖 凋亡 

分 类 号：R714.256[医药卫生—妇产科学]