lncRNA SNHG1靶向miR-145-5p/PDCD4轴对缺氧/复氧诱导的心肌细胞凋亡的影响

Effect of lncRNA SNHG1 targeting miR-145-5p/PDCD4 axis on hypoxia/reoxygenation-induced cardiomyocyte apoptosis

目的探讨长链非编码RNA(lncRNA)核内小RNA宿主基因1(SNHG1)靶向miR-145-5p/程序性细胞死亡因子4(PDCD4)轴对缺氧/复氧(H/R)诱导的心肌细胞凋亡的影响。方法将H9c2细胞分为对照组(NC组)、H/R组、si-NC组、si-SNHG1组、mimic NC组、miR-145-5p mimic组、si-SNHG1+inhibitor NC组、si-SNHG1+miR-145-5p inhibitor组。除NC组外,其他组H9c2细胞均需转染对应物质后构建H/R模型。实时定量PCR(qRT-PCR)检测H9c2细胞中SNHG1、miR-145-5p表达;CCK-8法、流式细胞术分别检测细胞增殖、凋亡;试剂盒检测H9c2细胞中超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量;Western blotting检测H9c2细胞中PDCD4、caspase 3、Bcl-2、Bax蛋白表达;双荧光素酶报告基因实验验证SNHG1与miR-145-5p、miR-145-5p与PDCD4的关系。结果沉默SNHG1或过表达miR-145-5p可促进H/R诱导的H9c2细胞中miR-145-5p表达和细胞增殖,抑制PDCD4蛋白表达、细胞凋亡和氧化应激。miR-145-5p inhibitor减弱了沉默SNHG1对H/R诱导的H9c2细胞中miR-145-5p表达和细胞增殖的促进作用,以及对PDCD4蛋白表达、细胞凋亡、氧化应激的抑制作用。SNHG1靶向调控miR-145-5p/PDCD4轴。结论沉默SNHG1可能通过调控miR-145-5p/PDCD4轴抑制H/R诱导的H9c2细胞凋亡。

Objective To explore the effect of long noncoding RNA(lncRNA)small nucleolar RNA host gene 1(SNHG1)targeting of the miR-145-5p/programmed cell death 4(PDCD4)axis on hypoxia/reoxygenation(H/R)-induced cardiomyocyte apoptosis.Methods H9c2 cells were divided into normal control(NC),H/R,si-NC,si-SNHG1,mimic NC,miR-145-5p mimic,si-SNHG1+inhibitor NC,and si-SNHG1+miR-145-5p inhibitor groups.H9c2 cells in all but the NC group were transfected with corresponding substances to construct the H/R model.Real-time quantitative polymerase chain reaction was used to detect SNHG1 and miR-145-5p expressions in H9c2 cells.A cell counting kit-8 assay and flow cytometry were used to detect cell proliferation and apoptosis,respectively.Superoxide dismutase activity and malondialdehyde content of H9c2 cells were detected using kits.Western blotting was used to detect the expressions of PDCD4,caspase 3,Bcl-2,and Bax protein in H9c2 cells.The relationship between the SNHG1 and miR-145-5p and between miR-145-5p and PDCD4 was verified in a double luciferase reporter gene experiment.Results SNHG1 silencing or miR-145-5p overexpression promoted H/R-induced miR-145-5p expression and proliferation in H9c2 cells and inhibited PDCD4 protein expression,apoptosis,and oxidative stress.An miR-145-5p inhibitor attenuated the promotive effect of SNHG1 silencing on H/R-induced miR-145-5p expression and proliferation in H9c2 cells and the inhibitory effect on PDCD4 protein expression,apoptosis,and oxidative stress.Thus,SNHG1 silencing targeted and regulated the miR-145-5p/PDCD4 axis.Conclusion SNHG1 gene silencing may inhibit H/R-induced H9c2 cell apoptosis by regulating the miR-145-5p/PDCD4 axis.