微小 RNA-325-3p 靶向人附睾蛋白 4 缓解肺动脉高压大鼠右心室纤维化。
MicroRNA-325-3p Targets Human Epididymis Protein 4 to Relieve Right Ventricular Fibrosis in Rats with Pulmonary Arterial Hypertension.
机构信息
Department of Cardiology, Hunan Provincial People's Hospital, The First Affiliated Hospital of Hunan Normal University, Clinical Medicine Research Center of Heart Failure of Hunan Province, Hunan Normal University, Changsha, Hunan 410005, China.
College of Pharmaceutical Science, Key Laboratory of Pharmaceutical Quality Control of Hebei Province, Hebei University, Baoding, Hebei 071002, China.
出版信息
Cardiovasc Ther. 2022 Jan 22;2022:4382999. doi: 10.1155/2022/4382999. eCollection 2022.
BACKGROUND
Pulmonary arterial hypertension (PAH) usually causes right ventricular dysfunction, which is closely related to cardiac fibrosis. But cardiac fibrosis mechanism remains unclear. Our purpose was to explore microRNA-325-3p (miR-325-3p)/human epididymis protein 4's (HE4) role in the occurrence and development of right ventricular fibrosis in PAH.
METHODS
The right ventricular fibrosis model of rats with PAH was constructed, and miR-325-3p was overexpressed to explore miR-325-3p's effect on myocardial fibrosis in rats with PAH. Pearson correlation coefficient examined the correlation between HE4 and miR-325-3p. We separated and identified the primary rat myocardial fibroblasts from the heart tissue. Then, the Ang II-treated myocardial fibroblast fibrosis model was constructed. miR-325-3p mimics and si-HE4 and oe-HE4 cell lines were constructed to investigate miR-325-3p and HE4 effects on myocardial cell fibrosis. Then, we added PI3K inhibitor LY294002 to study the effects of HE4 in cell fibrosis by the PI3K/AKT pathway. Starbase was used to predict miR-325-3p and HE4 binding sites. Dual-luciferase reporter assay verified whether miR-325-3p and HE4 were targeted.
RESULTS
Overexpression of miR-325-3p alleviated myocardial fibrosis in rats with PAH. Pearson correlation coefficient showed that HE4 was negatively correlated with miR-325-3p expression. Starbase predicted that miR-325-3p had binding sites with HE4. Dual-luciferase reporter assay demonstrated that miR-325-3p targeted HE4. Overexpression of miR-325-3p downregulated HE4 and inhibited myocardial fibroblast fibrosis. HE4 knockdown inhibited myocardial fibroblast fibrosis. HE4 promoted myocardial fibroblast fibrosis and activated the PI3K/AKT pathway. In addition, HE4 affected myocardial fibroblast fibrosis through the PI3K/AKT pathway.
CONCLUSIONS
miR-325-3p could target HE4 to relieve right ventricular fibrosis in rats with PAH. This study could provide new targets and strategies for right ventricular fibrosis in PAH.
背景
肺动脉高压（PAH）通常会导致右心室功能障碍，而这与心脏纤维化密切相关。然而，心脏纤维化的机制尚不清楚。我们的目的是探讨微小 RNA-325-3p（miR-325-3p）/人附睾蛋白 4（HE4）在 PAH 中右心室纤维化发生和发展中的作用。
方法
构建 PAH 大鼠右心室纤维化模型，过表达 miR-325-3p 以探讨 miR-325-3p 对 PAH 大鼠心肌纤维化的影响。Pearson 相关系数检测 HE4 与 miR-325-3p 的相关性。从心脏组织中分离并鉴定原代大鼠心肌成纤维细胞。然后，构建 Ang II 处理的心肌成纤维细胞纤维化模型。构建 miR-325-3p 模拟物和 si-HE4、oe-HE4 细胞系，研究 miR-325-3p 和 HE4 对心肌细胞纤维化的影响。然后，加入 PI3K 抑制剂 LY294002，通过 PI3K/AKT 通路研究 HE4 对细胞纤维化的影响。Starbase 用于预测 miR-325-3p 和 HE4 的结合位点。双荧光素酶报告基因实验验证 miR-325-3p 和 HE4 是否靶向结合。
结果
过表达 miR-325-3p 可减轻 PAH 大鼠的心肌纤维化。Pearson 相关系数显示，HE4 与 miR-325-3p 的表达呈负相关。Starbase 预测 miR-325-3p 与 HE4 有结合位点。双荧光素酶报告基因实验证实 miR-325-3p 靶向 HE4。过表达 miR-325-3p 下调 HE4 并抑制心肌成纤维细胞纤维化。HE4 敲低抑制心肌成纤维细胞纤维化。HE4 促进心肌成纤维细胞纤维化并激活 PI3K/AKT 通路。此外，HE4 通过 PI3K/AKT 通路影响心肌成纤维细胞纤维化。
结论
miR-325-3p 可靶向 HE4 缓解 PAH 大鼠的右心室纤维化。本研究可为 PAH 右心室纤维化提供新的靶点和策略。
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