Chen Yihui, Hong Shichai, Wang Zhefeng, Hong Xiang, Chen Gang, Huang Yulong, Lin Yue, Xie Xinsheng, Lin Chenwei, Lu Weifeng
Department of Vascular Surgery, Xiamen Branch, Zhongshan Hospital, Fudan University, Xiamen, China.
Biotherapy Center, Xiamen Branch, Zhongshan Hospital, Fudan University, Xiamen, China.
Protein Pept Lett. 2025;32(1):62-74. doi: 10.2174/0109298665347753241028072130.
This study aimed to explore whether excessive HIF2α can amplify the impact of human Umbilical Cord Mesenchymal Stem Cell-derived Extracellular Vesicles (hUC-MSC- EVs) on endothelial cells.
In this study, we created HIF2α-overexpressing hUC-MSC-EVs and compared their pro-angiogenic effects with control EVs on Human Umbilical Vein Endothelial Cells (HUVECs). MTT assay and Edu staining were used to detect the viability and proliferation ability of HUVECs, and Transwell and Tube Formation Assays were used to detect cell migration and tube formation ability. qPCR assay was used to detect the expression of cellular angiogenic markers. Subsequently, miRNAs that might be regulated by HIF2α were predicted by bioinformatics analysis, and qPCR was used to detect the relative expression of miRNAs in HUVECs treated with hUC-MSC- EV, which over-expresses HIF2α. Subsequently, miR-146a inhibitors were used to investigate the role of miR-146a in mediating the pro-angiogenic effect of HIF2α on HUVECs by detecting cell viability, proliferation, migration, tube-forming ability, and expression of angiogenic markers. Finally, AKT/ERK phosphorylation and Spred1 expression were detected using Western blotting.
Our findings have indicated that overexpression of HIF2α significantly enhances the ability of hUC-MSC-EVs to stimulate proliferation, migration, and tube formation in HUVECs, as demonstrated by MTT/Edu staining, Transwell assay, and tube formation assay results, respectively. Mechanistically, excessive HIF2α has been found to induce the expression of miR-146a in HUVECs and the overexpression of a miR-146a inhibitor to negate the influence of excessive HIF2α on hUC-MSC-EV-induced activity in HUVECs.
The overexpression of HIF2α is an effective strategy for enhancing the pro-angiogenic function of hUC-MSC-EVs.
本研究旨在探讨过量的低氧诱导因子2α（HIF2α）是否会增强人脐带间充质干细胞衍生的细胞外囊泡（hUC-MSC-EVs）对内皮细胞的影响。
在本研究中，我们构建了过表达HIF2α的hUC-MSC-EVs，并将其对人脐静脉内皮细胞（HUVECs）的促血管生成作用与对照细胞外囊泡进行比较。采用MTT法和Edu染色检测HUVECs的活力和增殖能力，采用Transwell法和管腔形成实验检测细胞迁移和管腔形成能力。采用qPCR法检测细胞血管生成标志物的表达。随后，通过生物信息学分析预测可能受HIF2α调控的微小RNA（miRNAs），并采用qPCR法检测过表达HIF2α的hUC-MSC-EV处理的HUVECs中miRNAs的相对表达。随后，使用miR-146a抑制剂，通过检测细胞活力、增殖、迁移、管腔形成能力和血管生成标志物的表达，研究miR-146a在介导HIF2α对HUVECs促血管生成作用中的作用。最后，使用蛋白质免疫印迹法检测AKT/ERK磷酸化和Sprouty相关蛋白1（Spred1）的表达。
我们的研究结果表明，HIF2α的过表达显著增强了hUC-MSC-EVs刺激HUVECs增殖、迁移和管腔形成的能力，MTT/Edu染色、Transwell实验和管腔形成实验结果分别证明了这一点。从机制上讲，已发现过量的HIF2α可诱导HUVECs中miR-146a的表达，而miR-146a抑制剂的过表达可消除过量HIF2α对hUC-MSC-EV诱导的HUVECs活性的影响。
HIF2α的过表达是增强hUC-MSC-EVs促血管生成功能的有效策略。
