PDF(Pubmed)
Abstract：
UNASSIGNED: It is essential to acknowledge that the cardiovascular toxicity associated with anthracycline drugs can be partially attributed to the damage inflicted on blood vessels and endothelial cells. Extracellular vesicles (EVs) derived from mesenchymal stem cells (MSCs) have the potential to repair cellular processes and promote tissue regeneration through the transfer of signaling molecules such as miRNAs. In the present study, we investigated the effects of MSC-EVs on daunorubicin (DNR)-damaged human cardiac microvascular endothelial cells (HCMEC) and developing blood vessels of Chicken Chorioallantoic Membrane (CAM) in vivo.
UNASSIGNED: We constructed in vitro and in vivo models of DNR-damaged endothelial cells and developing blood vessel. Scratch wound assays, EdU assays, tube formation assays, and SA-β-Gal staining were used to evaluate the effects of MSC-EVs on cell migration, proliferation, angiogenesis capacity and cell senescence. Blood vessel area was used to assess the effects of MSC-EVs on CAM vasculature. RT-qPCR was used to detect the mRNA expression levels of inflammatory molecules. RNA sequencing was employed to compare differential gene expression and downstream regulatory mechanisms. RNA interference experiments and miRNA mimic overexpression experiments were used to validate the regulatory effects of target genes and downstream signaling pathways.
UNASSIGNED: We found that MSC-EVs improved the migration, proliferation, and angiogenesis of HCMEC, while also alleviating cellular senescence. The angiogenic effect on the developing blood vessels was confirmed in vivo. We identified that MSC-EVs downregulated the expression of PARP9, thereby inhibiting the STAT1/pSTAT1 signaling pathway. This downregulation effect is likely mediated by the transfer of miR-186-5p from MSC-EVs to HCMEC. Overexpression of miR-186-5p in DNR-damaged HCMEC also exhibited the aforementioned downregulation effect. In vivo, the introduction of miR-186-5p mimics enhanced angiogenesis in the CAM model.
UNASSIGNED: To summarize, our study reveals that MSC-EVs can restore the cellular function of DNR-damaged HCMEC and alleviate cellular senescence through the miR-185-5p-PARP9-STAT1/pSTAT1 pathway. This finding highlights the potential of MSC-EVs as a therapeutic strategy for mitigating the detrimental effects of anthracycline-induced endothelial damage.
UNASSIGNED: We constructed in vitro and in vivo models of DNR-damaged endothelial cells and developing blood vessel. Scratch wound assays, EdU assays, tube formation assays, and SA-β-Gal staining were used to evaluate the effects of MSC-EVs on cell migration, proliferation, angiogenesis capacity and cell senescence. Blood vessel area was used to assess the effects of MSC-EVs on CAM vasculature. RT-qPCR was used to detect the mRNA expression levels of inflammatory molecules. RNA sequencing was employed to compare differential gene expression and downstream regulatory mechanisms. RNA interference experiments and miRNA mimic overexpression experiments were used to validate the regulatory effects of target genes and downstream signaling pathways.
UNASSIGNED: We found that MSC-EVs improved the migration, proliferation, and angiogenesis of HCMEC, while also alleviating cellular senescence. The angiogenic effect on the developing blood vessels was confirmed in vivo. We identified that MSC-EVs downregulated the expression of PARP9, thereby inhibiting the STAT1/pSTAT1 signaling pathway. This downregulation effect is likely mediated by the transfer of miR-186-5p from MSC-EVs to HCMEC. Overexpression of miR-186-5p in DNR-damaged HCMEC also exhibited the aforementioned downregulation effect. In vivo, the introduction of miR-186-5p mimics enhanced angiogenesis in the CAM model.
UNASSIGNED: To summarize, our study reveals that MSC-EVs can restore the cellular function of DNR-damaged HCMEC and alleviate cellular senescence through the miR-185-5p-PARP9-STAT1/pSTAT1 pathway. This finding highlights the potential of MSC-EVs as a therapeutic strategy for mitigating the detrimental effects of anthracycline-induced endothelial damage.
摘要：
■必须承认与蒽环类药物相关的心血管毒性可部分归因于对血管和内皮细胞造成的损害。源自间充质干细胞（MSC）的细胞外囊泡（EV）具有修复细胞过程并通过传递信号分子（例如miRNA）促进组织再生的潜力。在本研究中,我们在体内研究了MSC-EV对柔红霉素（DNR）损伤的人心脏微血管内皮细胞（HCMEC）和鸡绒毛尿囊膜（CAM）发育血管的影响。
■我们构建了DNR损伤的内皮细胞和发育血管的体外和体内模型。划痕伤口分析，EdU化验，管形成测定，和SA-β-Gal染色用于评估MSC-EV对细胞迁移的影响，扩散,血管生成能力和细胞衰老。血管面积用于评估MSC-EV对CAM脉管系统的影响。RT-qPCR用于检测炎症分子的mRNA表达水平。RNA测序用于比较差异基因表达和下游调控机制。RNA干扰实验和miRNA模拟物过表达实验用于验证靶基因和下游信号通路的调节作用。
■我们发现MSC-EV改进了迁移，扩散,和HCMEC的血管生成，同时也减轻了细胞衰老。在体内证实了对发育中的血管的血管生成作用。我们发现MSC-EV下调PARP9的表达，从而抑制STAT1/pSTAT1信号通路。这种下调效应可能是由miR-186-5p从MSC-EV转移到HCMEC介导的。miR-186-5p在DNR损伤的HCMEC中的过表达也表现出上述下调作用。在体内,miR-186-5p的引入模拟了CAM模型中血管生成的增强.
■总而言之，我们的研究表明,MSC-EV可通过miR-185-5p-PARP9-STAT1/pSTAT1通路恢复DNR损伤的HCMEC的细胞功能,缓解细胞衰老.这一发现强调了MSC-EV作为减轻蒽环类抗生素诱导的内皮损伤的有害影响的治疗策略的潜力。
■我们构建了DNR损伤的内皮细胞和发育血管的体外和体内模型。划痕伤口分析，EdU化验，管形成测定，和SA-β-Gal染色用于评估MSC-EV对细胞迁移的影响，扩散,血管生成能力和细胞衰老。血管面积用于评估MSC-EV对CAM脉管系统的影响。RT-qPCR用于检测炎症分子的mRNA表达水平。RNA测序用于比较差异基因表达和下游调控机制。RNA干扰实验和miRNA模拟物过表达实验用于验证靶基因和下游信号通路的调节作用。
■我们发现MSC-EV改进了迁移，扩散,和HCMEC的血管生成，同时也减轻了细胞衰老。在体内证实了对发育中的血管的血管生成作用。我们发现MSC-EV下调PARP9的表达，从而抑制STAT1/pSTAT1信号通路。这种下调效应可能是由miR-186-5p从MSC-EV转移到HCMEC介导的。miR-186-5p在DNR损伤的HCMEC中的过表达也表现出上述下调作用。在体内,miR-186-5p的引入模拟了CAM模型中血管生成的增强.
■总而言之，我们的研究表明,MSC-EV可通过miR-185-5p-PARP9-STAT1/pSTAT1通路恢复DNR损伤的HCMEC的细胞功能,缓解细胞衰老.这一发现强调了MSC-EV作为减轻蒽环类抗生素诱导的内皮损伤的有害影响的治疗策略的潜力。
