LncRNA TUG1 knockdown protects cardiomyocytes against hypoxia/reoxygenation-induced injury via regulating miR-532-5p/Sox8 axis

Xinyong Cai, Shu Wang, Lang Hong, Songping Yu, Bin Li, Hong Zeng, Xu Yang, Ping Zhang, Liang Shao
Journal of Cardiovascular Pharmacology 2020 August 19
Long non-coding RNA taurine upregulated gene 1 (lncRNA TUG1) has been reported to involve in the processing of cardiac ischemia/reperfusion (I/R) injury following myocardial infarction. Thus, this study further investigates the underlying mechanisms of TUG1 in hypoxia/reoxygenation (H/R)-induced cardiomyocyte injury in vitro. Cell viability, apoptosis, migration and invasion were detected using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, flow cytometry and transwell assay, respectively. Western blot was used to examine the levels of matrix metallopeptidase 9 (MMP-9), MMP2 and SRY-box transcription factor 8 (Sox8) protein. Levels of lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD), and glutathione peroxidase (GSH-PX) were detected using commercial kits. Levels of TUG1, microRNA-532-5p (miR-532-5p), and Sox8 were detected by quantitative real-time polymerase chain reaction (qRT-PCR). The interaction between miR-532-5p and Sox8 or TUG1 was confirmed by dual-luciferase reporter and RNA immunoprecipitation (RIP) assay. H/R induced rat cardiomyocyte H9c2 injury by inhibiting cell viability, migration and invasion, promoting cell apoptosis, and stimulating oxidative stress. H/R-induced up-regulated the level of TUG1, and TUG1 knockdown alleviated H/R-induced cardiomyocyte injury. TUG1 directly bound to miR-532-5p, and miR-532-5p inhibition reversed the action of TUG1 knockdown on H/R-induced cardiomyocyte injury. Sox8 was a target of miR-532-5p, and miR-532-5p blunted H/R-induced cardiomyocyte injury by targeting Sox8. Additionally, TUG1 knockdown inhibited H/R-induced Sox8 elevation via miR-532-5p in H9c2 cells. TUG1 silence ameliorated H/R-induced cardiomyocytes injury via regulating miR-532-5p/Sox8 axis, suggesting a potential therapeutic target for preventing myocardial ischaemia/reperfusion injury.

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