lncRNA Gm4419靶向miR-703对Aβ25-35诱导的PC12细胞氧化应激和凋亡的影响

Effects of lncRNA Gm4419 targeting miR-703 on Aβ25-35-induced oxidative stress and apoptosis in PC12 cells

目的探讨长链非编码RNA(lncRNA)Gm4419靶向miR-703对β-淀粉样蛋白(Aβ25-35)诱导的PC12细胞氧化应激和细胞凋亡的影响。方法Aβ25-35诱导PC12细胞损伤造模。流式细胞术检测细胞凋亡,酶联免疫吸附测定(ELISA)试剂盒检测超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量。实时定量PCR(RT-qPCR)分析Gm4419和miR-703表达量。采用上述方法检测Gm4419和miR-703表达对Aβ25-35诱导的PC12细胞氧化应激和细胞凋亡的影响。双荧光素酶报告基因实验和RT-qPCR确定Gm4419和miR-703靶向调控关系。结果Aβ25-35诱导后PC12细胞凋亡率、MDA含量、Gm4419表达显著升高(P<0.05),SOD活性、miR-703表达显著降低(P<0.05)。抑制Gm4419或过表达miR-703后Aβ25-35诱导的PC12细胞凋亡率、MDA含量显著降低(P<0.05),SOD活性显著升高(P<0.05)。miR-703是Gm4419的靶基因,Gm4419负调控miR-703表达。抑制miR-703表达逆转了Gm4419抑制对Aβ25-35诱导的PC12细胞氧化应激和细胞凋亡的影响(P<0.05)。结论抑制lncRNA Gm4419通过上调miR-703能够减轻Aβ25-35诱导的PC12细胞凋亡和氧化应激损伤。

Objective To investigate the effects of long non-coding RNA(lncRNA)Gm4419 targeting miR-703 onβ-amyloid(Aβ25-35)-induced oxidative stress and apoptosis in PC12 cells.Methods PC12 cells were treated with Aβ25-35 to establish cell injury model.Flow cytometry was used to detect cell apoptosis,and enzyme-linked immunosorbent assay(ELISA)kit was used to detect superoxide dismutase(SOD)activity and malondialdehyde(MDA)content.Real-time quantitative PCR(RT-qPCR)was applied to analyze the expression levels of Gm4419 and miR-703.The effects of Gm4419 and miR-703 expression on the oxidative stress and apoptosis of PC12 cells induced by Aβ25-35 were detected by the above methods.The dual luciferase reporter gene experiment and RT-qPCR were performed to confirm the target regulation relationship between Gm4419 and miR-703.Results After induction of Aβ25-35,the apoptosis rate,MDA content,and expression levels of Gm4419 in PC12 cells were significantly increased(P<0.05),whereas SOD activity and expression levels of miR-703 were significantly decreased(P<0.05).After inhibiting Gm4419 or overexpression of miR-703,the apoptosis rate and MDA content in PC12 cells induced by Aβ25-35 were significantly decreased(P<0.05),however,SOD activity was significantly increased(P<0.05).The miR-703 was the target gene of Gm4419,and Gm4419 negatively regulated the expression of miR-703.To inhibit miR-703 expression could reverse the effects of Gm4419 inhibition on the oxidative stress and apoptosis of PC12 cells induced by Aβ25-35(P<0.05).Conclusion Inhibiting lncRNA Gm4419 can alleviate Aβ25-35-induced apoptosis and oxidative stress damage in PC12 cells by up-regulating the expression levels of miR-703.