lncRNA PWAR5通过靶向miR-30a-5p/SOCS3分子轴降低心肌炎心肌细胞的损伤  

lncRNA PWAR5 reduces myocarditis cardiomyocyte damage by targeting miR-30a-5p/SOCS3 molecular axis

作　　者：卢英红 迟伟峰 谭玉婷 王春筱 纪文岩[1] 吴赛[1] Lu Yinghong;Chi Weifeng;Tan Yuting(Dept of Cardiology, Qingdao Haici Hospital, Qingdao University, Qingdao 266000)

机构地区：[1]青岛大学附属青岛市海慈医院心内科,青岛266000

出　　处：《安徽医科大学学报》2021年第12期1955-1959,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81900310)。

摘　　要：目的研究长链非编码(lncRNA)PWAR5在脂多糖诱导心肌炎时对心肌细胞的保护作用及分子机制。方法分离培养大鼠原代心肌细胞,分别采用载有PWAR5序列的慢病毒(PWAR5组)和空载慢病毒(Control组)进行感染,实时荧光定量PCR(qRT-PCR)检测感染效率。采用脂多糖诱导心肌炎后,MTS法检测PWAR5对大鼠原代心肌细胞活力的影响,ELISA法检测上清液中白细胞介素(IL)-1β和肿瘤坏死因子(TNF)-α的含量,流式细胞术分别检测PWAR5对大鼠原代心肌细胞凋亡的影响。分别采用生物信息学方法和双荧光素酶报告基因实验预测和验证PWAR5的靶基因。qRT-PCR检测靶基因的表达。Western blot法检测靶基因蛋白及凋亡相关蛋白的表达。结果与Control组相比,PWAR5组细胞中PWAR5表达增加(P<0.01)。脂多糖处理后,与Control组相比,PWAR5组细胞活力增加(P<0.01),上清液中IL-1β和TNF-α的含量下降(均P<0.01),PWAR5组细胞凋亡比例下降(P<0.01)。PWAR5的靶基因可能是miR-30a-5p,PWAR5可与miR-30a-5p互补结合(P<0.01)。与Control组相比,PWAR5组细胞中miR-30a-5p的表达下降(P<0.01),细胞因子信号传导抑制因子3(SOCS3)mRNA的表达增加(P<0.01)。与Control组相比,PWAR5组细胞中SOCS3和Bcl-2蛋白表达上升,Bax和Caspase-3蛋白表达下降。结论PWAR5可通过靶向调控miR-30a-5p/SOCS3分子轴,降低心肌炎导致的心肌细胞损伤。Objective To study the protective effect and molecular mechanism of long non-coding RNA(lncRNA)PWAR5 on cardiomyocytes in lipopolysaccharide-induced myocarditis.Methods The primary rat cardiomyocytes were isolated and cultured,and were infected with lentivirus carrying PWAR5 sequence(PWAR5 group)and empty lentivirus(Control group).Real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect infection efficiency.After lipopolysaccharide-induced myocarditis,the MTS method was used to detect the effect of PWAR5 on the viability of primary rat cardiomyocytes,and the ELISA method was used to detect the levels of interleukin(IL)-1βand tumor necrosis factor(TNF)-αin the supernatant.Flow cytometry was used to detect the effect of PWAR5 on the apoptosis of primary rat cardiomyocytes.Bioinformatics methods and dual-luciferase reporter gene experiments were used to predict and verify the target gene of PWAR5.qRT-PCR was used to detect the expression of target gene.Western blot was used to detect the expression of target gene protein and apoptosis-related proteins.Results Compared with the Control group,the expression of PWAR5 in the PWAR5 group increased(P<0.01).After lipopolysaccharide treatment,compared with the Control group,the cell viability of the PWAR5 group increased(P<0.01),the contents of IL-1βand TNF-αin the supernatant decreased(P<0.01),and the percentage of apoptosis in the PWAR5 group decreased(P<0.01).The target gene of PWAR5 might be miR-30a-5p,and PWAR5 could complement miR-30a-5p(P<0.01).Compared with the Control group,the expression of miR-30a-5p in the cells of the PWAR5 group decreased(P<0.01),and the expression of suppressor of cytokine signaling 3(SOCS3)mRNA increased(P<0.01).Compared with the Control group,the expression of SOCS3 and Bcl-2 protein in the PWAR5 group increased,while the expression of Bax and Caspase-3 protein decreased.Conclusion PWAR5 can reduce the damage of cardiomyocytes caused by myocarditis by targeting the miR-30a-5p/SOCS3 molecular axis.

关 键 词：心肌炎 PWAR5 miR-30a-5p SOCS3 细胞凋亡 

分 类 号：R542.2[医药卫生—心血管疾病]