恩格列净预处理的间充质干细胞来源的小细胞外囊泡减轻心脏损伤。

OBJECTIVE: Small extracellular vesicles derived from mesenchymal stem cells (MSCs) play important roles in cardiac protection. Studies have shown that the cardiovascular protection of sodium-glucose cotransporter 2 inhibitor (SGLT2i) is independent of its hypoglycemic effect. This study is aimed at investigating whether small extracellular vesicles derived from MSCs pretreated with empagliflozin (EMPA) has a stronger cardioprotective function after myocardial infarction (MI) and to explore the underlying mechanisms. METHODS AND RESULTS: We evaluated the effects of EMPA on MSCs and the effects of EMPA-pretreated MSCs-derived small extracellular vesicles (EMPA-sEV) on myocardial apoptosis, angiogenesis, and cardiac function after MI in vitro and in vivo. The small extracellular vesicles of control MSCs (MSC-sEV) and EMPA-pretreated MSCs were extracted, respectively. Small extracellular vesicles were cocultured with apoptotic H9c2 cells induced by HO or injected into the infarcted area of the Sprague-Dawley (SD) rat myocardial infarction model. EMPA increased the cell viability, migration ability, and inhibited apoptosis and senescence of MSCs. In vitro, EMPA-sEV inhibited apoptosis of H9c2 cells compared with the control group (MSC-sEV). In the SD rat model of MI, EMPA-sEV inhibited myocardial apoptosis and promoted angiogenesis in the infarct marginal areas compared with the MSC-sEV. Meanwhile, EMPA-sEV reduced infarct size and improved cardiac function. Through small extracellular vesicles (miRNA) sequencing, we found several differentially expressed miRNAs, among which miR-214-3p was significantly elevated in EMPA-sEV. Coculture of miR-214-3p high expression MSC-derived small extracellular vesicles with H9c2 cells produced similar protective effects. In addition, miR-214-3p was found to promote AKT phosphorylation in H9c2 cells. CONCLUSIONS: Our data suggest that EMPA-sEV significantly improve cardiac repair after MI by inhibiting myocardial apoptosis. miR-214-3p at least partially mediated the myocardial protection of EMPA-sEV through the AKT signaling pathway.

目的：间充质干细胞（MSCs）衍生的小细胞外囊泡在心脏保护中发挥重要作用。研究表明，钠-葡萄糖共转运蛋白 2 抑制剂（SGLT2i）的心血管保护作用与其降血糖作用无关。本研究旨在探讨经恩格列净（EMPA）预处理的 MSCs 衍生的小细胞外囊泡（EMPA-sEV）在心肌梗死（MI）后是否具有更强的心脏保护功能，并探讨其潜在机制。

方法和结果：我们评估了 EMPA 对 MSCs 的影响，以及 EMPA 预处理的 MSC 衍生的小细胞外囊泡（EMPA-sEV）对 MI 后心肌细胞凋亡、血管生成和心功能的影响。分别提取对照组 MSCs（MSC-sEV）和 EMPA 预处理的 MSCs 的小细胞外囊泡。小细胞外囊泡与 HO 诱导的凋亡 H9c2 细胞共培养，或注射到 Sprague-Dawley（SD）大鼠 MI 模型的梗死区。EMPA 增加了 MSCs 的细胞活力、迁移能力，并抑制了细胞凋亡和衰老。在体外，与对照组（MSC-sEV）相比，EMPA-sEV 抑制了 H9c2 细胞的凋亡。在 SD 大鼠 MI 模型中，与 MSC-sEV 相比，EMPA-sEV 抑制了梗死边缘区的心肌细胞凋亡并促进了血管生成。同时，EMPA-sEV 减少了梗死面积并改善了心功能。通过小细胞外囊泡（miRNA）测序，我们发现了几种差异表达的 miRNA，其中 miR-214-3p 在 EMPA-sEV 中显著升高。与高表达 miR-214-3p 的 MSC 衍生的小细胞外囊泡共培养的 H9c2 细胞产生了类似的保护作用。此外，还发现 miR-214-3p 促进了 H9c2 细胞中 AKT 的磷酸化。

结论：我们的数据表明，EMPA-sEV 通过抑制心肌细胞凋亡，显著改善 MI 后的心脏修复。miR-214-3p 通过 AKT 信号通路至少部分介导了 EMPA-sEV 的心肌保护作用。