长链非编码 RNA HOTAIR 通过上调 RAGE 促进急性心肌梗死后的炎症反应。
HOTAIR promotes inflammatory response after acute myocardium infarction by upregulating RAGE.
机构信息
Department of Cardiology, Taizhou People's Hospital, Taizhou, China.
出版信息
Eur Rev Med Pharmacol Sci. 2018 Nov;22(21):7423-7430. doi: 10.26355/eurrev_201811_16282.
OBJECTIVE
The aim of this study was to explore the role of HOTAIR in inflammatory response after acute myocardium infarction (AMI) and to investigate its underlying mechanism.
MATERIALS AND METHODS
The AMI model was first constructed in rats, and heart tissues were harvested. Expression levels of HOTAIR and receptor of advanced glycation end-products (RAGE) in rat heart were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The protein expression level of pEKR in rat heart was detected by Western blot. The levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in rats were determined by enzyme-linked immunosorbent assay (ELISA). The hypoxia-induced H9C2 cells were used to construct the MI model in vitro. Meanwhile, the expression levels of HOTAIR and RAGE in H9C2 cells were detected. The levels of TNF-α and IL-6 in the culture medium were determined by ELISA. Rescue experiments were conducted by co-transfecting pcDNA-HOTAIR and si-RAGE in H9C2 cells. Subsequently, the levels of pERK, TNF-α, and IL-6 were detected.
RESULTS
The mRNA expression levels of HOTAIR and RAGE in the AMI group were significantly higher than those of the control group. Western blot showed remarkably higher protein levels of RAGE and pERK in AMI rats when compared with those of controls. Similarly, results of ELISA indicated that the levels of TNF-α and IL-6 in AMI rats were significantly higher than those of controls. Meanwhile, overexpression of HOTAIR in H9C2 cells remarkably elevated the expression levels of HOTAIR and RAGE. In addition, upregulated pERK, TNF-α, and IL-6 were observed in H9C2 cells overexpressing HOTAIR, which could be reversed by RAGE knockdown.
CONCLUSIONS
HOTAIR promotes inflammatory response after AMI by upregulating RAGE expression.
目的
本研究旨在探讨 HOTAIR 在急性心肌梗死（AMI）后炎症反应中的作用，并探讨其潜在机制。
材料和方法
首先在大鼠中构建 AMI 模型，然后采集心脏组织。通过定量实时聚合酶链反应（qRT-PCR）检测大鼠心脏中 HOTAIR 和晚期糖基化终产物受体（RAGE）的表达水平。通过 Western blot 检测大鼠心脏中 pEKR 的蛋白表达水平。通过酶联免疫吸附试验（ELISA）测定大鼠肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）的水平。体外使用缺氧诱导的 H9C2 细胞构建 MI 模型。同时，检测 H9C2 细胞中 HOTAIR 和 RAGE 的表达水平。通过 ELISA 测定培养基中 TNF-α和 IL-6 的水平。在 H9C2 细胞中转染 pcDNA-HOTAIR 和 si-RAGE 进行挽救实验。然后检测 pERK、TNF-α和 IL-6 的水平。
结果
AMI 组 HOTAIR 和 RAGE 的 mRNA 表达水平明显高于对照组。Western blot 显示 AMI 大鼠 RAGE 和 pERK 的蛋白水平明显高于对照组。同样，ELISA 结果表明 AMI 大鼠 TNF-α和 IL-6 的水平明显高于对照组。此外，H9C2 细胞中 HOTAIR 的过表达显著提高了 HOTAIR 和 RAGE 的表达水平。此外，在过表达 HOTAIR 的 H9C2 细胞中观察到上调的 pERK、TNF-α和 IL-6，通过 RAGE 敲低可逆转这一现象。
结论
HOTAIR 通过上调 RAGE 表达促进 AMI 后的炎症反应。
Zotero 插件