lncRNA SNHG14通过miR-181c-5p/SOX6信号轴调控缺血性脑卒中神经元细胞凋亡

LncRNA SNHG14 regulates neuronal apoptosis in ischemic stroke through miR-181c-5p/SOX6 signal axis

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。

Abstract:

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.