LncRNA ROR modulates myocardial ischemia-reperfusion injury mediated by the miR-185-5p/CDK6 axis.

Abstract

LncRNAs and miRNAs are correlated with the pathogenesis of myocardial ischemia-reperfusion injury (MIRI). Whether lncRNA ROR or miR-185-5p plays a crucial role in MIRI is still unclear. In in-vitro, human cardiac myocytes (HCMs) were treated with hypoxia/reoxygenation (H/R). Wistar rats were used to set up an in-vitro I/R model by means of recanalization after ligation. Evaluation of the myocardial injury marker lactate dehydrogenase (LDH) in HCMs cells was performed. The expression of miR-185-5p and ROR, IL-1β, and IL-18 were detected by qRT-PCR. ELISA was also performed to evaluate the secretion of IL-1β and IL-18. Western blotting was carried out to determine CDK6, NLRP3, GSDMD-N, ASC, and cleaved-caspase1 protein expression. The relationship between miR-185-5p and CDK6 or ROR was confirmed by a dual-luciferase reporter assay. Our findings revealed that H/R treated HCMs showed a significantly decreased miR-185-5p expression and increased expression of CDK6 and ROR. ROR knockdown reduced H/R induced pyroptosis and inflammation, while knockdown of miR-185-5p accelerated the effect. Furthermore, miR-185-5p was negatively regulated and absorbed by ROR in HCMs. Overexpression of miR-185-5p reversed the H/R-induced cell pyroptosis and upregulation of LDH, IL-1β, and IL-18. In HCMs, miR-185-5p was also negatively regulated and related to CDK6 expression. Moreover, overexpression of CDK6 significantly inhibited the effects of miR-185-5p mimics on the inflammatory response and pyroptosis of HCMs. Knockdown of ROR alleviated H/R-induced myocardial injury by elevating miR-185-5p and inhibiting CDK6 expression. Taken together, our results show that the ROR/miR-185-5p/CDK6 axis modulates cell pyroptosis induced by H/R and the inflammatory response of HCMs.

• Authors: Jing Sun, Yan-Meng Zhu, Qin Liu, Yan-Hui Hu, Chang Li, Huan-Huan Jie, Guo-Hai Xu, Ren-Jie Xiao, Xian-Liang Xing, Shu-Chun Yu, Ying-Ping Liang
• Journal: Laboratory investigation; a journal of technical methods and pathology (Lab Invest) Vol. 102 Issue 5 Pg. 505-514 (05 2022) ISSN: 1530-0307 [Electronic] United States
• PMID: 35066566 (Publication Type: Journal Article)
• Copyright: © 2022. The Author(s), under exclusive licence to United States and Canadian Academy of Pathology.

Chemical References

• Interleukin-18
• MIRN185 microRNA, rat
• MicroRNAs
• RNA, Long Noncoding

Topics

• Animals
• Hypoxia
• Interleukin-18
• MicroRNAs (genetics, metabolism)
• Myocardial Reperfusion Injury (genetics)
• RNA, Long Noncoding (genetics)
• Rats
• Rats, Wistar