Kang Hui, Yu Hui, Zeng Ling, Ma Hao, Cao Ge
Department of Cardiovascular Surgery, Sichuan University West China Hospital, Chengdu, Sichuan Province, China.
Department of Critical Care Medicine, Sichuan University West China Hospital, Chengdu, Sichuan Province, China.
Hypertens Res. 2022 Jun;45(6):976-989. doi: 10.1038/s41440-022-00884-6. Epub 2022 Mar 9.
Myocardial ischemia-reperfusion injury (MIRI) is a pathological process characterized by cardiomyocyte death. Long noncoding RNAs (lncRNAs) have been shown to be dysregulated in the course of MIRI. Accordingly, the current study investigated the mechanism of lncRNA Rian in MIRI-induced cardiomyocyte pyroptosis. First, a murine model of MIRI was established by using the left anterior descending (LAD) coronary artery ligation method. Cardiac function and myocardial histopathological changes were evaluated by echocardiography and hematoxylin and eosin staining. Then, a cell model of MIRI was established by oxygen-glucose deprivation/reoxygenation (OGD/R), followed by analysis of NLRP3, cleaved caspase-1, and GSDMD-N levels by western blotting. The levels of IL-1β, IL-18, TNF-α, and IL-10 were measured using ELISA. LncRNA Rian, miR-17-5p, and CCND1 expression in myocardial tissues and OGD/R cells were examined using RT-qPCR. Finally, the binding relationships between Rian and miR-17-5p and miR-17-5p and CCND1 were validated with the help of dual-luciferase and RNA pull-down assays. Rian was poorly expressed in MIRI mice and OGD/R cells. LncRNA Rian overexpression reduced cardiomyocyte pyroptosis in vivo and in vitro, as indicated by decreased NLRP3, cleaved caspase-1, GSDMD-N, IL-1β, IL-18, and TNF-α levels and increased IL-10 levels. Furthermore, Rian bound to miR-17-5p and promoted CCND1 transcription. Notably, miR-17-5p overexpression or CCND1 silencing reversed the inhibitory effect of Rian overexpression on cardiomyocyte pyroptosis. Collectively, our findings indicate that Rian overexpression reduces cardiomyocyte pyroptosis and alleviates MIRI through the miR-17-5p/CCND1 axis.
心肌缺血再灌注损伤（MIRI）是一种以心肌细胞死亡为特征的病理过程。长链非编码RNA（lncRNAs）已被证实在MIRI过程中表达失调。因此，本研究探讨了lncRNA Rian在MIRI诱导的心肌细胞焦亡中的作用机制。首先，采用左冠状动脉前降支（LAD）结扎法建立MIRI小鼠模型。通过超声心动图和苏木精-伊红染色评估心脏功能和心肌组织病理学变化。然后，通过氧糖剥夺/复氧（OGD/R）建立MIRI细胞模型，随后通过蛋白质免疫印迹法分析NLRP3、裂解的caspase-1和GSDMD-N水平。使用酶联免疫吸附测定法检测IL-1β、IL-18、TNF-α和IL-10水平。采用逆转录定量聚合酶链反应（RT-qPCR）检测心肌组织和OGD/R细胞中lncRNA Rian、miR-17-5p和CCND1的表达。最后，借助双荧光素酶和RNA下拉试验验证Rian与miR-17-5p以及miR-17-5p与CCND1之间的结合关系。Rian在MIRI小鼠和OGD/R细胞中低表达。lncRNA Rian过表达在体内和体外均减少了心肌细胞焦亡，表现为NLRP3、裂解的caspase-1、GSDMD-N、IL-1β、IL-18和TNF-α水平降低，IL-10水平升高。此外，Rian与miR-17-5p结合并促进CCND1转录。值得注意的是，miR-17-5p过表达或CCND1沉默可逆转Rian过表达对心肌细胞焦亡的抑制作用。总之，我们的研究结果表明，Rian过表达通过miR-17-5p/CCND1轴减少心肌细胞焦亡并减轻MIRI。
