| 何梦,孙健,沈巨信,秦娥.CircRNA-Tbpl1海绵吸附miR-214-3p调控自噬在低氧性肺动脉高压血管重塑中的作用及其机制何梦,孙健,沈巨信,秦娥[J].浙江中西医结合杂志,2024,34(5): |
| CircRNA-Tbpl1海绵吸附miR-214-3p调控自噬在低氧性肺动脉高压血管重塑中的作用及其机制何梦,孙健,沈巨信,秦娥 |
| The role and mechanism of circRNA-Tbpl1 sponge adsorbing miR-214-3p to regulate autophagy in vascular remodeling in hypoxic pulmonary hypertension |
| 投稿时间：2022-11-04 修订日期：2024-04-18 |
| DOI： |
| 中文关键词: 低氧性肺动脉高压 CircRNA-Tbpl1 自噬 miR-214-3p 血管重塑 |
| 英文关键词:HPH CircRNA-Tbpl1 Autophagy miR-214-3p Vascular remodeling |
| 基金项目:基金项目：1. 浙江省自然科学基金探索项目，LQ20H010001；2. 绍兴市卫生健康科技计划卫生健康科技计划项目，2022KY006； |
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| 中文摘要: |
| 目的 探讨circRNA-Tbpl1通过海绵吸附miR-214-3p调控自噬在低氧性肺动脉高压血管重塑中的作用,并分析其机制。方法 构建低氧性肺动脉高压(hypoxic pulmonary hypertension, HPH)小鼠模型,分离小鼠原代肺动脉平滑肌细胞(PASMCs)。低氧处理PASMCs,CCK-8检测细胞活性,Western blot检测自噬相关蛋白表达,qRT-PCR检测circRNA-Tbpl1的相对表达水平。通过转染和自噬诱导剂雷帕霉素(rapamycin, Rap)干预,分析circRNA-Tbpl1与PASMCs自噬的相关性。通过共转染sh-circRNA-Tbpl1和miR-214-3p抑制物,分析circRNA-Tbpl1/ miR-214-3p对低氧诱导的PASMCs自噬和细胞增殖活性的影响。结果 与对照组小鼠相比,HPH小鼠的右心室收缩压(right ventricular systolic pressure, RVSP)、右心室肥厚指数(right ventricular hypertrophy index, RVHI)显著增加,且HPH小鼠肺血管出现明显的中膜肥大和外膜增生,明显出现肺血管重塑。HPH小鼠的肺组织和低氧处理的PASMCs中LC3 II、Beclin-1表达水平显著上调,p62表达下调,circRNA-Tbpl1水平升高。下调circRNA-Tbpl1可抑制低氧诱导的PASMCs自噬,减弱细胞增殖活性。双荧光素酶报告基因检测结果显示circRNA-Tbpl1海绵吸附miR-214-3p,miR-214-3p在HPH小鼠和低氧诱导的PASMCs中表达降低,与circRNA-Tbpl1呈负相关。与sh-NC组相比,sh-circRNA-Tbpl1组的自噬水平减弱,细胞增殖活力降低。与sh-circRNA-Tbpl1 + miRNA-NC组相比,sh-circRNA-Tbpl1 + miR-214-3p抑制物组的自噬水平升高,细胞增殖活力增强。结论 circRNA-Tbpl1通过海绵吸附miR-214-3p抑制低氧诱导的自噬,从而缓解低氧性肺动脉高压血管重塑。 |
| 英文摘要: |
| Objective To investigate the role of circRNA-Tbpl1 in regulating autophagy in vascular remodeling in hypoxic pulmonary hypertension through sponge adsorption of miR-214-3p, and to analyze its mechanism. Methods A hypoxic pulmonary hypertension (HPH) mouse model was constructed, and mouse primary pulmonary artery smooth muscle cells (PASMCs) were isolated. PASMCs were treated with hypoxia, the cell viability was detected by CCK-8, the expression of autophagy-related proteins was detected by Western blot, and the relative expression level of circRNA-Tbpl1 was detected by qRT-PCR. The correlation between circRNA-Tbpl1 and autophagy in PASMCs was analyzed by transfection and the intervention of autophagy inducer rapamycin (Rap). By co-transfection of sh-circRNA-Tbpl1 and miR-214-3p inhibitor, the effect of circRNA-Tbpl1/miR-214-3p on hypoxia-induced autophagy and cell proliferation activity of PASMCs was analyzed. Results Compared with control mice, right ventricular systolic pressure (RVSP) and right ventricular hypertrophy index (RVHI) were significantly increased in HPH mice, and pulmonary blood vessels in HPH mice were significantly increased. Medial hypertrophy and adventitial hyperplasia, with marked pulmonary vascular remodeling. The expression levels of LC3 II and Beclin-1 were significantly up-regulated, the expression of p62 was down-regulated, and the level of circRNA-Tbpl1 was up-regulated in the lung tissue and hypoxia-treated PASMCs of HPH mice. Downregulation of circRNA-Tbpl1 inhibited hypoxia-induced autophagy in PASMCs and attenuated cell proliferation activity. The results of dual-luciferase reporter gene assay showed that circRNA-Tbpl1 sponge adsorbed miR-214-3p, and the expression of miR-214-3p was decreased in HPH mice and hypoxia-induced PASMCs, which was negatively correlated with circRNA-Tbpl1. Compared with the sh-NC group, the sh-circRNA-Tbpl1 group had attenuated levels of autophagy and decreased cell proliferation activity. Compared with the sh-circRNA-Tbpl1 + miRNA-NC group, the sh-circRNA-Tbpl1 + miR-214-3p inhibitor group had higher levels of autophagy and enhanced cell proliferation activity. Conclusion circRNA-Tbpl1 inhibits hypoxia-induced autophagy by sponge-adsorbing miR-214-3p, thereby alleviating vascular remodeling in hypoxic pulmonary hypertension. |
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