CircACTR2调节miR-23a-3p/TBL1X轴对高糖诱导的滋养层细胞损伤的影响  

Influence of circACTR2 on high glucose-induced trophoblast cell injury by regulating miR-23a-3p/TBL1X axis

作　　者：黄春艳 向汨 费志医 高琴 HUANG Chunyan;XIANG Mi;FEI Zhiyi;GAO Qin(Department of Endocrinology,Puren Hospital,Wuhan 430082,China)

机构地区：[1]武汉市普仁医院内分泌科,430082

出　　处：《天津医药》2023年第10期1048-1054,共7页Tianjin Medical Journal

基　　金：湖北省卫生计生委中医药科研项目(ZY2019F036)。

摘　　要：目的探讨环状RNA肌动蛋白相关蛋白2(circACTR2)调节miR-23a-3p/转导素β1X连锁蛋白(TBL1X)轴对高糖诱导的滋养层细胞损伤的影响。方法将人绒毛膜滋养层细胞HTR-8/Svneo分为NG组(5.5 mmol/L葡萄糖)、HG组(25 mmol/L葡萄糖)、si-NC组(25 mmol/L葡萄糖+转染si-NC)、si-circACTR2组(25 mmol/L葡萄糖+转染si-circACTR2)、si-circACTR2+inhibitor-NC组(25 mmol/L葡萄糖+si-circACTR2和inhibitor-NC共转染)、sicircACTR2+miR-23a-3p inhibitor组(25 mmol/L葡萄糖+si-circACTR2和miR-23a-3p inhibitor共转染)。实时荧光定量PCR(qPCR)检测细胞中circACTR2、miR-23a-3p的表达;CCK-8法检测细胞增殖;流式细胞仪检测细胞凋亡;划痕实验检测细胞迁移;酶联免疫吸附试验检测丙二醛(MDA)水平和乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)活性;Western blot检测细胞中TBL1X、增殖细胞核抗原(PCNA)、基质金属蛋白酶(MMP)-2、MMP-9、胱天蛋白酶3(caspase-3)的表达。双萤光素酶报告基因实验分别验证circACTR2、TBL1X和miR-23a-3p的靶向关系。结果与NG组相比,HG组HTR-8/Svneo细胞miR-23a-3p表达、增殖能力、划痕愈合率、PCNA、MMP-2、MMP-9表达、SOD活性降低,circACTR2、TBL1X和caxpase-3表达、MDA含量、LDH活性、凋亡率升高(P<0.05);与HG组和si-NC组相比,si-circACTR2组HTR-8/Svneo细胞中miR-23a-3p表达、增殖能力、划痕愈合率、PCNA、MMP-2、MMP-9表达、SOD活性升高,circACTR2、TBL1X和caxpase-3表达、MDA含量、LDH活性、凋亡率降低(P<0.05);在敲低circACTR2的基础上,下调miR-23a-3p可明显减弱circACTR2敲低对高糖诱导的HTR-8/Svneo细胞的增殖和迁移的促进作用,增强细胞凋亡和氧化应激能力。双萤光素酶报告基因实验结果显示,circACTR2靶向负调控miR-23a-3p表达,miR-23a-3p靶向负调控TBL1X表达。结论敲低circACTR2可调控miR-23a-3p/TBL1X轴,进而通过抑制高糖诱导的滋养层细胞损伤来发挥保护作用。Objective To investigate the influence of circular RNA actin related protein 2(circACTR2)on high glucose-induced trophoblast cell injury by regulating miR-23a-3p/transducinβ1X-linked protein(TBL1X)axis.Methods Human chorionic trophoblast cells HTR-8/Svneo were grouped into the NG group(5.5 mmol/L glucose),the HG group(25 mmol/L glucose),the si-NC group(25 mmol/L glucose+transfected with si-NC),the si-circACTR2 group(25 mmol/L glucose+transfected with si-circACTR2),the si-circACTR2+inhibitor-NC group(25 mmol/L glucose+co transfected with si-circACTR2 and inhibitor-NC)and si-circACTR2+miR-23a-3p inhibitor group(25 mmol/L glucose+co transfected with si-circACTR2 and miR-23a-3p inhibitor).Real-time quantitative PCR(qPCR)was performed to measure expression levels of circACTR2 and miR-23a-3p in cells.Cell proliferation was detected by CCK-8 assay.Cell apoptosis was detected by flow cytometry.Scratch assay was performed to detect cell migration.Enzyme-linked immunosorbent assay was applied to detect malondialdehyde(MDA)level,lactate dehydrogenase(LDH)and superoxide dismutase(SOD)activities.Western blot assay was used to detect expressions of TBL1X,proliferating cell nuclear antigen(PCNA),matrix metalloproteinase-2(MMP-2),MMP-9 and cysteine protease-3(caspase-3)in cells.Dual-luciferase reporter gene experiment was performed to verify the targeting relationship between circACTR2,TBL1X and miR-23a-3p respectively.Results Compared with the NG group,the HTR-8/Svneo cell expression of miR-23a-3p,proliferative ability,scratch healing rate,expression of PCNA,MMP-2,MMP-9 and SOD activity were reduced in the HG group,and the expression of circACTR2,TBL1X,caxpase-3,MDA content,LDH activity and apoptosis rate were increased(P<0.05).Compared with the HG group and the si-NC group,the miR-23a-3p expression,proliferative ability,scratch healing rate,expression of PCNA,MMP-2 and MMP-9,and SOD activity in HTR-8/Svneo cells were increased in the si-circACTR2 group,and the expression of circACTR2,TBL1X and caxpase-3,MDA content,LDH activi

关 键 词：肌动蛋白相关蛋白质2 妊娠特异性β1糖蛋白质类 滋养层 细胞凋亡 miR-23a-3p 高糖 人绒毛膜滋养层细胞HTR-8/Svneo 

分 类 号：R349.64[医药卫生—基础医学]