Human umbilical cord mesenchymal stem cells derived exosomes exert antiapoptosis effect via activating PI3K/Akt/mTOR pathway on H9C2 cells
Human umbilical cord mesenchymal stem cells derived exosomes exert antiapoptosis effect via activating PI3K/Akt/mTOR pathway on H9C2 cells
复制标题
人脐带间充质干细胞来源的外泌体通过激活PI3K/Akt/mTOR通路对H9C2细胞发挥抗凋亡作用
DOI：
10.1002/jcb.28705
复制
发表时间：
2019-09-01
影响因子：
4
通讯作者：
Wang, Guogan
中科院分区：
文献类型：
--
作者：
Liu, Hui;Sun, Xiaolu;Wang, Guogan
背景/目的近年来，外显子作为一种替代干细胞治疗心血管疾病（CVD）的方法引起了研究者的广泛关注。本研究旨在探讨人脐带间充质干细胞（UC-MSCs）衍生外泌体在H9C2细胞凋亡中的作用及其可能机制。方法从正常UC-MSCs培养基和缺氧预处理培养基中分离外泌体。透射电镜观察外泌体的形态。采用纳米颗粒跟踪分析检测外泌体的大小分布和浓度。采用Western blot分析外泌体表面标记物CD63。采用低氧和血清剥夺（H/SD）诱导H9C2细胞凋亡，然后分组处理。细胞计数试剂盒-8检测H9C2细胞活力。Hochest染色及annexin V-FITC/PI检测细胞凋亡。Western blot分析细胞凋亡、自噬、PI3K/Akt/mTOR通路相关蛋白的表达水平。免疫荧光法检测LC3B的表达。结果缺氧预处理使UC-MSCs外泌体分泌增加。UC-MSCs来源的外泌体可抑制H/ sd诱导的H9C2细胞凋亡。缺氧预处理增强了UC-MSCs的抗凋亡作用。缺氧预处理UC-MSCs衍生的外泌体（H-Exo）下调LC3B-II/I和beclin-1，上调P62、p-Akt/Akt和p-mTOR/mTOR。LY294002和雷帕霉素可减弱H-Exo的抗凋亡作用。结论UC-MSCs衍生外泌体可通过PI3K/Akt/mTOR通路调控自噬，抑制H/SD诱导的H9C2细胞凋亡。缺氧预处理可通过增加UC-MSCs外泌体分泌来增强上述作用。
Background/Objectives In recent years, as an alternative to stem cell therapy for cardiovascular diseases (CVD), exsomes have attracted wide attention among researchers. The present study aimed to investigate the role of human umbilical cord mesenchymal stem cells (UC-MSCs) derived exosomes play on H9C2 cells apoptosis and possible mechanisms. Methods Exosomes were isolated from normal UC-MSCs culture media and hypoxic preconditioning culture media. Transmission electron microscopy was used to observe the morphology of exosomes. Nanoparticle tracking analysis was used to detect the size distribution and concentration of exosomes. Western blot analysis was used to analyzed the surface marker CD63 of exosomes. H9C2 cells were induced apoptosis by hypoxia and serum deprivation (H/SD) and then were treated respectively by group. Cell Counting Kit-8 assay was used to detect viability of H9C2 cells. Apoptosis was detected by Hochest staining and annexin V-FITC/PI. The expression levels of related proteins of apoptosis, autophagy, and PI3K/Akt/mTOR pathway were analyzed by Western blot analysis. Immunofluorescence was used to analyze LC3B expression. Results Hypoxic preconditioning increased the exosomes secretion of UC-MSCs. UC-MSCs derived exosomes could inhibit H/SD-induced H9C2 cells apoptosis. Hypoxic preconditioning strengthened this antiapoptosis effect of UC-MSCs. Hypoxic preconditioning UC-MSCs derived exosomes (H-Exo) downregulated LC3B-II/I and beclin-1 and upregulated P62, p-Akt/Akt and p-mTOR/mTOR. The antiapoptotic effect of H-Exo could be attenuated by treatment with LY294002 and rapamycin. Conclusion UC-MSCs derived exosomes could inhibit H9C2 cells apoptosis induced by H/SD through regulating autophagy via PI3K/Akt/mTOR pathway. Hypoxia preconditioning could enhance above effects through increasing exosomes secretion of UC-MSCs.