miR-129-5p靶向SOX6对氧糖剥夺/复氧诱导的神经细胞凋亡及氧化应激的影响  

Effect of miR-129-5p on Neuronal Apoptosis and Oxidative Stress Induced by Oxygen Glucose Deprivation/Reoxygenation by Targeting SOX6

作　　者：霍江涛 蔡津津 梁惠 郭功兵 张小乔 HUO Jiangtao;CAI Jinjin;LIANG Hui;GUO Gongbing;ZHANG Xiaoqiao(Department of Geriatrics,Taihe Hospital,Shiyan City,Hubei Province(Affiliated Hospital of Hubei University of Medicine),Shiyan 442000,China)

机构地区：[1]湖北省十堰市太和医院(湖北医药学院附属医院),老年医学科,十堰442000

出　　处：《中国细胞生物学学报》2024年第12期2073-2080,共8页Chinese Journal of Cell Biology

摘　　要：该研究旨在探究微小RNA-129-5p(miR-129-5p)靶向SRY-box转录因子6(SOX6)对氧糖剥夺/复氧(OGD/R)诱导的神经细胞凋亡及氧化应激的影响。将人神经元细胞(SH-SY5Y细胞)分为Control组、OGD/R组、miR-NC组、miR-129-5p mimics组、miR-129-5p mimics+pcDNA-NC组和miR-129-5p mimics+pcDNA-SOX6组,除Control组外,其余组细胞均进行OGD/R诱导。采用RTqPCR检测细胞中miR-129-5p和SOX6 mRNA表达量;CCK-8试剂盒检测细胞增殖率;流式细胞术检测细胞凋亡情况;ELISA试剂盒检测细胞内超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GPx)含量;H2DCFDA荧光探针检测细胞活性氧(ROS)水平;Western blot检测细胞中SOX6、Caspase-3和NF-κB蛋白表达情况。研究结果得出,与Control组比较,OGD/R组细胞中miR129-5p表达水平、细胞活性、SOD、CAT和GPx活性下降(P<0.05),SOX6 mRNA表达水平、细胞凋亡率、ROS水平、SOX6、Caspase-3和p-NF-κB/NF-κB蛋白表达水平上升(P<0.05);与OGD/R组比较,miR-129-5p mimics组细胞miR-129-5p表达水平、细胞活性、SOD、CAT和GPx活性升高(P<0.05),SOX6 mRNA表达水平、细胞凋亡率、ROS水平、SOX6、Caspase-3和p-NF-κB/NF-κB蛋白表达水平降低(P<0.05);与miR-129-5p mimics组比较,miR-129-5p mimics+pcDNA-SOX6组细胞miR-129-5p表达水平、细胞活性、SOD、CAT和GPx活性下降(P<0.05),SOX6 mRNA表达水平、细胞凋亡率、ROS水平、SOX6和p-NF-κB/NF-κB蛋白表达水平上升(P<0.05)。双荧光素酶报告基因实验证实SOX6是miR-129-5p的靶基因。因此,过表达miR-129-5p可下调SOX6,从而减轻OGD/R诱导的神经细胞凋亡及氧化应激。The purpose of this study was to investigate the effect of microRNA-129-5p(miR-129-5p)targeting SOX6(SRY-box transcription factor 6)on OGD/R(oxoglucose deprivation/reoxygenation)-induced apoptosis and oxidative stress in neuronal cells.Human neuronal cells(SH-SY5Y cells)were separated into control group,OGD/R group,miR-NC group,miR-129-5p mimics group,miR-129-5p mimics+pcDNA-NC group,and miR-129-5p mimics+pcDNA-SOX6 group.Except for the control group,all other groups were induced with OGD/R.RT-qPCR was applied to detect the expression levels of miR-129-5p and SOX6 mRNA in cells.CCK-8 assay kit was used to detect cell proliferation rate.Flow cytometry was used to detect cell apoptosis.ELISA kit was used to detect the levels of intracellular SOD(superoxide dismutase),CAT(catalase),and GPx(glutathione peroxidase).H2DCFDA fluorescent probe was applied to detect cellular ROS(reactive oxygen species)level.Western blot was applied to detect the expression of SOX6,Caspase-3,and NF-κB proteins in cells.The results of the study are derived compared with the control group,the expression of miR-129-5p,cell activity,SOD,CAT,and GPx activities in OGD/R group decreased(P<0.05),while the expression of SOX6 mRNA,apoptosis rate,level of ROS,the expression of SOX6,Caspase-3,and p-NF-κB/NF-κB proteins increased(P<0.05).Compared with the OGD/R group,the expression of miR-129-5p,cell activity,SOD,CAT,and GPx activities in miR-129-5p mimics group increased(P<0.05),while the expression of SOX6 mRNA,apoptosis rate,level of ROS,the expression of SOX6,Caspase-3,and p-NF-κB/NF-κB proteins decreased(P<0.05).Compared with the miR-129-5p mimics group,the expression of miR-129-5p,cell activity,SOD,CAT,and GPx activities in miR-129-5p mimics+pcDNA-SOX6 group decreased(P<0.05),while the expression of SOX6 mRNA,apoptosis rate,level of ROS,the expression of SOX6 and p-NF-κB/NF-κB proteins increased(P<0.05).Dual luciferase reporter gene experiment confirmed that SOX6 was the target gene of miR129-5p.Therefore,overexpression of miR-129-5p can

关 键 词：微小RNA-129-5p SRY-box转录因子6 氧糖剥夺/复氧诱导 神经细胞凋亡 氧化应激 

分 类 号：R743.3[医药卫生—神经病学与精神病学]