Department of Hepatopancreatobiliary Surgery, The first Affiliated Hospital of Anhui Medical University, Hefei, Anhui, China.
Cell Cycle. 2019 Dec;18(24):3491-3501. doi: 10.1080/15384101.2019.1689480. Epub 2019 Nov 10.
The purpose of this study was to explore the associated mechanism by which MSCs-derived exosomes exerted protective effect in hepatic ischemia/reperfusion injury (IRI). Human umbilical cord blood mesenchymal stem cells (hUCB-MSCs)-derived exosomes were administrated into LO2 cells exposed to hypoxia/reoxygenation (H/R) and mice subjected to IRI. Cell viability was assessed by CCK-8 assay. Apoptosis was analyzed by flow cytometry and TUNEL staining. The expression of miR-1246 and Wnt/β-catenin pathway-related proteins was detected by quantitative real-time PCR (qRT-PCR) and western blotting. The concentration of pro-inflammatory cytokines was determined by ELISA. Luciferase activity assay was performed to confirm the interaction between miR-1246 and glycogen synthase kinase 3β (GSK3β). Hepatic function was assessed by determining serum alanine amino transferase (ALT) and aspartate amino transferase (AST) levels. Histological changes were observed using hematoxylin-eosin (H&E) staining. MiR-1246 was significantly downregulated in H/R-treated LO2 cells. Treatment with exosomes derived from hUCB-MSCs led to miR-1246 upregulation. Furthermore, hUCB-MSCs-derived exosomes induced anti-apoptotic and pro-survival effects in LO2 cells and ameliorated IRI-induced hepatic dysfunction in mice, while treatment of exosomes from miR-1246 inhibitor-transfected hUCB-MSCs showed opposite effect, which was mediated by regulating GSK3β-Wnt/β-catenin pathway. Collectively, hUCB-MSCs-derived exosomes alleviated hepatic IRI by transporting miR-1246 via regulating GSK3β-mediated Wnt/β-catenin pathway.
本研究旨在探讨间充质干细胞衍生的外泌体在肝缺血/再灌注损伤（IRI）中发挥保护作用的相关机制。将人脐血间充质干细胞（hUCB-MSCs）衍生的外泌体给予缺氧/复氧（H/R）处理的 LO2 细胞和 IRI 小鼠。通过 CCK-8 测定法评估细胞活力。通过流式细胞术和 TUNEL 染色分析细胞凋亡。通过定量实时 PCR（qRT-PCR）和蛋白质印迹法检测 miR-1246 和 Wnt/β-catenin 通路相关蛋白的表达。通过酶联免疫吸附测定（ELISA）测定促炎细胞因子的浓度。通过荧光素酶活性测定法证实 miR-1246 与糖原合成酶激酶 3β（GSK3β）之间的相互作用。通过测定血清丙氨酸氨基转移酶（ALT）和天冬氨酸氨基转移酶（AST）水平评估肝功能。通过苏木精-伊红（H&E）染色观察组织学变化。H/R 处理的 LO2 细胞中 miR-1246 显著下调。用 hUCB-MSCs 衍生的外泌体处理导致 miR-1246 上调。此外，hUCB-MSCs 衍生的外泌体在 LO2 细胞中诱导抗凋亡和促生存作用，并改善 IRI 诱导的小鼠肝功能障碍，而用 miR-1246 抑制剂转染的 hUCB-MSCs 衍生的外泌体处理则显示相反的作用，这是通过调节 GSK3β-Wnt/β-catenin 通路介导的。总之，hUCB-MSCs 衍生的外泌体通过调节 GSK3β 介导的 Wnt/β-catenin 通路转运 miR-1246 来减轻肝 IRI。
