LncRNA KCNQ1OT1调节miR-20a-5p/PTEN轴对OGD/R诱导的海马神经元损伤的影响OA
Effect of Long Non-Coding RNA KCNQ1OT1 on Oxygen-Glucose Deprivation/Reoxygenation(OGD/R)-Induced Hippocampal Neuronal Damage by Regulating miR-20a-5p/PTEN Axis
目的 探讨长链非编码RNA(LncRNA)KCNQ1OT1调节微小RNA(miR)-20a-5p/磷酸酶和张力蛋白同源物(PTEN)轴对糖氧剥夺/复糖复氧(OGD/R)诱导的海马神经元细胞损伤的影响.方法 将HT22细胞分为对照组、OGD/R组、si-NC 组、si-KCN 组、miR mimic-NC 组、miR-20a-5p mimic 组、si-KCN+miR inhibitor-NC 组、si-KCN+miR-20a-5p inhibitor组.MTT法检测HT22细胞活力;试剂盒检测细胞中ROS、SOD、TNF-α、IL-1β和IL-6水平;流式细胞术检测细胞凋亡;RT-qPCR实验检测细胞中LncRNA KCNQ1OT1、miR-20a-5p和PTEN mRNA水平;Western Blot测定PTEN和增殖相关蛋白表达;双荧光素酶报告基因试验分析miR-20a-5p与LncRNA KCNQ1OT1和PTEN的靶向作用.结果 与对照组相比,OGD/R 组 HT22 细胞存活率、SOD、miR-20a-5p、Ki67 和 PCNA 蛋白表达水平显著降低(P＜0.05),ROS、TNF-α、IL-1β、IL-6、细胞凋亡率、LncRNA KCNQ1OT1、PTEN mRNA及蛋白表达水平显著升高(P＜0.05);与si-NC或miR mimic-NC组相比,si-KCN 或 miR-20a-5p mimic 组细胞存活率、SOD、miR-20a-5p、Ki67 和 PCNA 蛋白表达水平显著升高(P＜0.05),ROS、TNF-α、IL-1β、IL-6、细胞凋亡率、LncRNA KCNQ1OT1、PTEN mRNA及蛋白表达水平显著降低(P＜0.05);相较于si-KCN+miR inhibitor-NC 组,si-KCN+miR-20a-5p inhibitor 组细胞存活率、SOD、miR-20a-5p、Ki67 和 PCNA 水平显著降低(P＜0.05),ROS、TNF-α、IL-1β、IL-6、细胞凋亡率和 PTEN mRNA 及蛋白表达水平显著升高(P＜0.05);与 miR mimic-NC 和 KC-NQ1OT1-WT或PTEN-WT共转染的细胞相比,miR-20a-5p mimic和KCNQ1OT1-WT或PTEN-WT共转染的细胞中相对荧光素酶活性显著减少(P＜0.05).结论 下调LncRNA KCNQ1OT1可能通过上调miR-20a-5p,降低PTEN表达,促进O GD/R诱导的海马神经元细胞增殖,抑制细胞氧化应激和炎症损伤.
Objective To investigate the effect of long non-coding RNA(LncRNA)KCNQ1OT1 on oxygen-glucose dep-rivation/reoxygenation(OGD/R)-induced hippocampal neuronal cell damage by regulating microRNA(miR)-20a-5p/phosphatase and tensin homologue deleted on chromosome ten(PTEN)axis.Methods HT22 cells were assigned into control group,OGD/R group,si-NC,si-KCN,miR-20a-5p mimic-NC,OGD/R+miR-20a-5p mimic,si-KCN+miR inhibitor-NC group,and si-KCN+miR-20a-5p inhibitor group.MTT assay was used to detect the viability of HT22 cells.Reagent kits were used to detect ROS,SOD,TNF-α,IL-1β,and IL-6 in cells.Flow cytometry was used to detect cell apoptosis.RT-qPCR were used to detect the mRNA levels of LncRNA KCNQ1OT1,miR-20a-5p,and PTEN mRNA in cells.Western blot experiments were used to detect PTEN and cell pro-liferation related proteins in cells.Dual-luciferase reporter gene assay was used to determine the targeting relationship between miR-20a-5p and LncRNA KCNQ1OT1,as well as between miR-20a-5p and PTEN.Results Compared with control group,OGD/R group exhibited a significantly reduced HT22 cell survival rate,as well as decreased SOD,miR-20a-5p,Ki67,and PCNA proteins(P＜0.05),while showing significantly increased ROS,TNF-α,IL-1β,IL-6,cell apoptosis rate,LncRNA KCNQ1OT1,PTEN mR-NA,and protein(P＜0.05).Compared with si-NC group or miR mimic-NC group,si-KCN or miR-20a-5p mimic group had a sig-nificantly increased cell survival rate,along with increased SOD,miR-20a-5p,Ki67,and PCNA proteins(P＜0.05),and signifi-cantly decreased ROS,TNF-α,IL-1β,IL-6,cell apoptosis rate,LncRNA KCNQ1OT1,PTEN mRNA,and protein(P＜0.05).Com-pared with si-KCN+miR inhibitor-NC group,si-KCN+miR-20a-5p inhibitor group had a significantly reduced cell survival rate,SOD,miR-20a-5p,Ki67,and PCNA(P＜0.05),and significantly increased ROS,TNF-α,IL-1β,IL-6,cell apoptosis rate,PTEN mRNA,and protein(P＜0.05).In the cells co-transfected with miR-20a-5p mimic and KCNQ1OT1-WT or PTEN-WT,the relative luciferase activity was significantly reduced(P＜0.05).Conclusion Downregulation of LncRNA KCNQ1OT1 may promote OGD/R-induced hippocampal neuronal cell proliferation,inhibit cellular oxidative stress and inflammatory damage by upregulating miR-20a-5p and reducing PTEN.
邱志雄;薛飞;王龙;顾伟杰;史轩丰
江阴市中医院急诊科,江苏江阴 214400江阴市中医院急诊科,江苏江阴 214400江阴市中医院急诊科,江苏江阴 214400江阴市中医院急诊科,江苏江阴 214400江阴市中医院急诊科,江苏江阴 214400
医药卫生
长链非编码RNA KCNQ1OT1微小RNA-20a-5p/磷酸酶和张力蛋白同源物轴海马神经元
long non coding RNA KCNQ1OT1microRNA-20a-5p/phosphatase and tensin homologue deleted on chromo-some ten axishippocampal neurons
《四川医学》 2026 (3)
279-286,8
江阴市卫生健康委科研项目(编号:M202214)
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