长链非编码 RNA H19 通过海绵吸附 miR-193b-3p 调节平滑肌细胞功能并参与主动脉夹层的发生。
LncRNA H19 regulates smooth muscle cell functions and participates in the development of aortic dissection through sponging miR-193b-3p.
机构信息
Department of Cardiovascular Surgery, Peking University Shenzhen Hospital, 1120 Lianhua Road, Shenzhen City, Guangdong Province, PR China.
Department of Vascular Surgery, Shanghai Changhai Hospital, Shanghai, PR China.
出版信息
Biosci Rep. 2021 Jan 29;41(1). doi: 10.1042/BSR20202298.
BACKGROUND
Multiple studies showed that long-chain noncoding RNA H19 (LncRNA H19) is high-expressed in human and mouse abdominal aortic aneurysms (AAAs). We speculated that it plays an important role in arterial disease, and therefore studied the role and mechanism of H19 in aortic dissection (AD).
METHODS
The expressions of related genes in human aortic smooth muscle cells (HASMCs) induced by platelet-derived growth factor BB (PDGF-BB) or in the aortic tissue of AD patients/mice were identified by Western blot and quantitative real-time polymerase chain reaction. The targeting relationship between H19 and miR-193b-3p was predicted and verified by bioinformatics analysis, dual luciferase assay, RNA pull-down assay, RNA immunoprecipitation (RIP), and Pearson correlation coefficient. The H19 and miR-193b-3p effects on the biological functions of tissues and cells were examined by MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide, thiazolyl blue tetrazolium bromide) assay, wound-healing assay, and Hematoxylin-Eosin (HE) staining.
RESULTS
LncRNA H19 was abnormally high-expressed in thoracic aorta tissues of AD patients, and it could competitively bind to and inhibit miR-193b-3p. In the PDGF-BB group, the expressions of H19, matrix metallopeptidase (MMP) 2 (MMP-2) and MMP-9 were up-regulated and the expressions of miR-193b-3p, α-SMA, and SM22α were down-regulated; moreover, the proliferation and migration rate of HASMCs were increased. However, H19 silencing reversed the regulation of PDGF-BB on HASMCs. More interestingly, miR-193b-3p inhibitor could partially reverse the effect of H19 silencing. In addition, the above results were verified by animal experiments, showing that shH19 and up-regulated miR-193b-3p could significantly reduce the thoracic aorta pathological damage in AD mice.
CONCLUSION
LncRNA H19 regulated smooth muscle cell function by sponging miR-193b-3p and it participated in the development of AD.
背景
多项研究表明，长链非编码 RNA H19（LncRNA H19）在人和鼠的腹主动脉瘤（AAA）中高表达。我们推测它在动脉疾病中发挥重要作用，因此研究了 H19 在主动脉夹层（AD）中的作用和机制。
方法
通过 Western blot 和实时定量聚合酶链反应鉴定血小板衍生生长因子 BB（PDGF-BB）诱导的人主动脉平滑肌细胞（HASMCs）或 AD 患者/小鼠主动脉组织中相关基因的表达。通过生物信息学分析、双荧光素酶报告基因实验、RNA 下拉实验、RNA 免疫沉淀（RIP）和 Pearson 相关系数验证 H19 与 miR-193b-3p 的靶向关系。通过 MTT（噻唑蓝比色法）实验、划痕实验和苏木精-伊红（HE）染色检测 H19 和 miR-193b-3p 对组织和细胞生物学功能的影响。
结果
AD 患者胸主动脉组织中 LncRNA H19 异常高表达，可竞争性结合并抑制 miR-193b-3p。在 PDGF-BB 组中，H19、基质金属蛋白酶（MMP）2（MMP-2）和 MMP-9 的表达上调，miR-193b-3p、α-SMA 和 SM22α 的表达下调；此外，HASMCs 的增殖和迁移率增加。然而，沉默 H19 逆转了 PDGF-BB 对 HASMCs 的调节作用。更有趣的是，miR-193b-3p 抑制剂可部分逆转 H19 沉默的作用。此外，动物实验验证了上述结果，表明 shH19 和上调的 miR-193b-3p 可显著减轻 AD 小鼠的胸主动脉病理损伤。
结论
LncRNA H19 通过海绵吸附 miR-193b-3p 调节平滑肌细胞功能，参与 AD 的发生发展。
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