Shanxi Medical University, Taiyuan, Shanxi, China (mainland).
Department of Cardiology, The Second Hospital of Shanxi Medical University, Taiyuan, Shanxi, China (mainland).
Med Sci Monit. 2020 Aug 9;26:e924016. doi: 10.12659/MSM.924016.
BACKGROUND The objective of the study was to explore the role of long non-coding RNA SNHG8 (lncRNA SNHG8) in myocardial infarction (MI) and the related mechanism of action. MATERIAL AND METHODS In vitro model of MI was established by hypoxia induction in cardiomyocyte line H9c2 cells. H9c2 cells were transfected with control-plasmid, SNHG8-plasmid, control-shRNA and SNHG8-shRNA. Quantitative real-time polymerase chain reaction (qRT-PCR) assay was performed to measure transfection efficiency. Creatine kinase-muscle/brain (CK-MB) release, cardiac troponin 1 (cTnI) release and mitochondria viability were detected by using related detection kits. MTT (3-(45)-dimethylthiahiazo (-z-y 1)-35-diphenytetrazoliumromide) assay was used to detect cell viability and flow cytometry analysis was used to detect cell apoptosis. Western blot assay was performed to measure protein expression of cleaved-Caspase3, p-p65 and p65. Enzyme-linked immunosorbent assay (ELISA) and qRT-PCR assay were performed to detect expression of interleukin (IL)-1ß, tumor necrosis factor (TNF)-alpha and IL-6. RESULTS LncRNA SNHG8 was overexpressed in hypoxia-induced cardiomyocytes. SNHG8-plasmid increased lncRNA SNHG8 expression, CK-MB release, cTnI release, and mitochondria viability in hypoxia-induced H9c2 cells. In addition, SNHG8-plasmid reduced cell viability, induced cell apoptosis, and increased expression of cleaved-caspase3, IL-1ß, TNF-alpha, IL-6, and p-p65 in hypoxia-induced H9c2 cells, while the effects of SNHG8-shRNA were opposite. CONCLUSIONS We demonstrated that lncRNA SNHG8 affected myocardial infarction by affecting hypoxia-induced cardiomyocyte injury via regulation of the NF-kappaB pathway.
本研究旨在探讨长链非编码 RNA SNHG8（lncRNA SNHG8）在心肌梗死（MI）中的作用及其相关作用机制。
通过诱导心肌细胞系 H9c2 细胞缺氧建立 MI 的体外模型。将对照质粒、SNHG8 质粒、对照 shRNA 和 SNHG8 shRNA 转染 H9c2 细胞。采用实时定量聚合酶链反应（qRT-PCR）检测转染效率。采用相关检测试剂盒检测肌酸激酶-肌肉/脑（CK-MB）释放、心肌肌钙蛋白 1（cTnI）释放和线粒体活力。MTT（3-(45)-dimethylthiahiazo (-z-y 1)-35-diphenytetrazoliumromide）法检测细胞活力，流式细胞术检测细胞凋亡。Western blot 法检测裂解型 Caspase3、p-p65 和 p65 蛋白表达。酶联免疫吸附试验（ELISA）和 qRT-PCR 法检测白细胞介素（IL）-1β、肿瘤坏死因子（TNF）-α和 IL-6 的表达。
在缺氧诱导的心肌细胞中，lncRNA SNHG8 表达上调。SNHG8 质粒增加了缺氧诱导的 H9c2 细胞中 lncRNA SNHG8 的表达、CK-MB 释放、cTnI 释放和线粒体活力。此外，SNHG8 质粒降低了细胞活力，诱导了细胞凋亡，并增加了缺氧诱导的 H9c2 细胞中 cleaved-caspase3、IL-1β、TNF-α、IL-6 和 p-p65 的表达，而 SNHG8 shRNA 的作用则相反。
我们证明，lncRNA SNHG8 通过调节 NF-κB 通路影响缺氧诱导的心肌细胞损伤，从而影响心肌梗死。
