摘要
目的通过IS体外模型探究lncRNA SNHG14在神经元细胞凋亡中的作用机制。方法通过氧葡萄糖剥夺(OGD)诱导的神经元细胞损伤来模拟IS。实时荧光定量聚合酶链反应检测SNHG14、miR-181c-5p、SOX6基因表达,CCK-8法检测细胞活力,流式细胞术检测细胞凋亡,Caspase-3检测试剂盒测定Caspase-3活性,RNA免疫沉淀实验和萤光素酶报告分析实验验证miR-181c-5p与SNHG14、SOX6的相互作用。结果sh-SNHG14组SNHG14 mRNA相对表达量低于sh-NC组(P<0.05),OGD+sh-SNHG14组SNHG14 mRNA相对表达量低于OGD+sh-NC组低(P<0.05)。OGD+sh-SNHG14组细胞活性率较OGD+sh-NC组高(P<0.05)。OGD+sh-SNHG14组细胞凋亡率、Caspase-3相对表达量较OGD+sh-NC组低(P<0.05)。miR-NC组miR-181c-5p相对表达量较miR-181c-5p mimics组低(P<0.05),inhibitor NC组较miR-181c-5p inhibitor组高(P<0.05)。SNHG14-WT+miR-181c-5p mimics组萤光素酶相对活性较SNHG14-WT+miR-NC组低(P<0.05),SNHG14-WT+miR-181c-5p inhibitor组较SNHG14-WT+inhibitor NC组高(P<0.05)。SOX6-WT+miR-181c-5p mimics组萤光素酶相对活性较SOX6-WT+miR-NC组低(P<0.05),SOX6-WT+miR-181c-5p inhibitor组较SOX6-WT+inhibitor NC组高(P<0.05)。miR-NC组SOX6 mRNA相对表达量较miR-181c-5p mimics组高(P<0.05),miR-181c-5p inhibitor组较inhibitor NC组高(P<0.05)。对照组细胞活性率较OGD+sh-SNHG14组高(P<0.05),OGD+sh-SNHG14+miR-181c-5p inhibitor组较OGD组高(P<0.05)。OGD+sh-SNHG14+miR-181c-5p inhibitor组细胞凋亡率较OGD+sh-SNHG14组高(P<0.05)。OGD+sh-SNHG14+miR-181c-5p inhibitor组Caspase-3相对表达量较OGD+sh-SNHG14组高(P<0.05)。结论SNHG14可调节IS模型中神经元细胞凋亡,作用机制可能是通过靶向miR-181c-5p/SOX6信号通路实现的。
Objective Explore the mechanism underlying the role of SNHG14 in neuronal apoptosis through an in vitro model of IS.Methods IS was simulated by oxygen glucose deprivation(OGD)-induced neuronal damage.The expressions of SNHG14,miR-181c-5p and SOX6 were detected via qPCR.The CCK8 assay was performed to measure the cell viability,while the apoptosis was determined via flow cytometry.The activity of caspase-3 was determined via a kit,and RIP and luciferase assays were applied to verify the interactions between miR-181c-5p and SNHG14 or SOX6.Results The relative expression of SNHG14 in the sh-SNHG14 group and the OGD+sh-SNHG14 group was lower than that in the sh-NC group and the OGD+sh-NC group,respectively(P<0.05).The cell viability in the OGD+sh-SNHG14 group was higher than that in the OGD+sh-NC group(P<0.05).The cell apoptosis rate and the relative expression of caspase-3 in the OGD+sh-SNHG14 group were lower than those in the OGD+sh-NC group(P<0.05).The relative expression of miR-181c-5p in the miR-NC group was lower than that in the miR-181c-5p mimics group(P<0.05),while that in the inhibitor NC group was higher than that in the miR-181c-5p inhibitor group(P<0.05).The relative luciferase activity in the SNHG14-WT+miR-181c 5p mimics group was lower than that in the SNHG14-WT+miR-NC group(P<0.05),while that in the SNHG14-WT+miR-181c-5p inhibitor group was higher than that in the SNHG14-WT+inhibitor NC group(P<0.05).The relative luciferase activity in the SOX6-WT+miR-181c-5p mimics group was lower than that in the SOX6-WT+miR-NC group(P<0.05),while that in the SOX6-WT+miR-181c-5p inhibitor group was higher than that in the SOX6-WT+inhibitor NC group(P<0.05).The relative expression of SOX6 mRNA in the miR-NC group was higher than that in the miR-181c-5p mimics group(P<0.05),and that in the miR-181c-5p inhibitor group was also higher than that in the inhibitor NC group(P<0.05).The cell viability in the control group was higher than that in the OGD+sh-SNHG14 group(P<0.05),while that in the OGD+sh-SNHG14+miR-181c-5p inhibitor group was higher than that in the OGD group(P<0.05).In contrast,the cell apoptosis rate in the OGD+sh-SNHG14+miR 181c-5p inhibitor group was higher than that in the OGD+sh-SNHG14 group(P<0.05).Besides,the relative expression of caspase-3 in the OGD+sh-SNHG14+miR-181c-5p inhibitor group was higher than that in the OGD+sh-SNHG14 group(P<0.05).Conclusions SNHG14 regulates neuronal apoptosis in IS models,at least partially through targeting the miR-181c-5p/SOX6 signaling pathway.
作者
钱旭东
李国芸
卜一
张硕
王红梅
窦志杰
Qian Xu-dong;Li Guo-yun;Bu Yi;Zhang Shuo;Wang Hong-mei;Dou Zhi-jie(Department of Neurology,Affiliated Hospital of Chengde Medical University,Chengde,Hebei 067000,China;Department of Respiratory Medicine,Affiliated Hospital of Chengde Medical University,Chengde,Hebei 067000,China)
出处
《中国现代医学杂志》
CAS
北大核心
2023年第12期41-48,共8页
China Journal of Modern Medicine
基金
河北省医学科学研究课题(No:20220005)。
