Department of Cardiology, Zhongshan Hospital, Fudan University, Shanghai Institute of Cardiovascular Diseases, 200032, Shanghai, China.
Department of Cardiology, Zoucheng Hospital, Affiliated Hospital of Jining Medical University, 273500, Jining, Shandong, China.
Curr Pharm Des. 2021;27(26):2966-2974. doi: 10.2174/1381612826666201005152709.
Angiogenesis is the most important repair process of tissues subjected to ischemic injury. The present study aims to investigate whether the pro-angiogenic effect of Qiliqiangxin prescription (QL) is mediated through miR-21 signaling.
Cardiac microvascular endothelial cells (CMECs) were isolated and cultured from 2-3 weeks old SD rats by the method of planting myocardium tissues. The purity was identified by CD31 immunofluorescence staining. CMECs were then cultured under 1% O2 hypoxia or normoxia condition for 24h in the presence or absence of QL pretreatment (QL, 0.5mg/ml, 24h). The mimics and inhibitors of miR-21 were transfected into CMECs. miR-21, HIF-1α, and VEGF expressions of CMECs were then detected by qRT-PCR and/or Western blot. The proliferation, migration, and tube formation functions of CMECs were assessed using the BrdU assay, wound healing test, and tube formation assay, respectively.
The results showed that compared with the control group, hypoxia significantly upregulated the expression of miR-21 and impaired CMECs proliferation, migration, and tube formation functions. Compared with the hypoxia group, QL further upregulated miR-21, HIF-1α, and VEGF expressions, and improved cell proliferation, migration, and tube formation of hypoxic CMECs. These effects of QL were abolished by a knockdown of miR-21. Conversely, treatment with miR-21 mimics further enhanced QL induced changes in hypoxic CMECs.
Results indicate that the pro-angiogenesis effects of QL on hypoxic CMECs are mediated by activating miR-21 and its downstream HIF-1α/VEGF pathway possibly.
血管生成是组织对缺血性损伤进行修复的最重要过程。本研究旨在探讨芪苈强心方的促血管生成作用是否通过 miR-21 信号通路介导。
采用心肌组织贴壁法，从 2-3 周龄 SD 大鼠心脏中分离培养心脏微血管内皮细胞（CMECs）。采用 CD31 免疫荧光染色法鉴定纯度。将 CMECs 在 1%O2 缺氧或常氧条件下培养 24 小时，同时给予或不给予 QL 预处理（QL，0.5mg/ml，24 小时）。将 miR-21 的模拟物和抑制剂转染到 CMECs 中。采用 qRT-PCR 和/或 Western blot 检测 CMECs 中 miR-21、HIF-1α 和 VEGF 的表达。采用 BrdU 检测试剂盒、划痕愈合试验和管形成试验分别评估 CMECs 的增殖、迁移和管形成功能。
结果表明，与对照组相比，缺氧显著上调 miR-21 的表达，并损害 CMECs 的增殖、迁移和管形成功能。与缺氧组相比，QL 进一步上调 miR-21、HIF-1α 和 VEGF 的表达，并改善缺氧 CMECs 的细胞增殖、迁移和管形成功能。miR-21 的敲低消除了 QL 的这些作用。相反，用 miR-21 模拟物处理进一步增强了 QL 诱导的缺氧 CMECs 的变化。
结果表明，QL 对缺氧 CMECs 的促血管生成作用可能是通过激活 miR-21 及其下游 HIF-1α/VEGF 通路介导的。
