Exosomes derived from mesenchymal stem cells inhibit mitochondrial dysfunction-induced apoptosis of chondrocytes via p38, ERK, and Akt pathways
Exosomes derived from mesenchymal stem cells inhibit mitochondrial dysfunction-induced apoptosis of chondrocytes via p38, ERK, and Akt pathways
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间充质干细胞来源的外泌体通过 p38、ERK 和 Akt 途径抑制线粒体功能障碍诱导的软骨细胞凋亡
DOI：
10.1007/s11626-019-00330-x
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发表时间：
2019-03-01
影响因子：
2.1
通讯作者：
Jin, Shao-Feng
中科院分区：
文献类型：
--
作者：
Qi, Hui;Liu, Dan-Ping;Jin, Shao-Feng
骨关节炎（OA）是世界范围内最常见的慢性关节疾病。软骨细胞作为软骨中唯一的常驻细胞，其凋亡在OA的发病中具有重要意义。间充质干细胞（MSC）为基础的治疗已被证明是有效的OA在动物和临床研究。目前，基于MSC的治疗的再生潜力主要归因于其旁分泌，其中外泌体可能发挥重要作用。在本研究中，我们的目的是找出MSC衍生的exosomes（MSC-Exos）的软骨细胞在正常和炎症条件下的活力的意义。体外培养兔骨髓间充质干细胞（BMSCs）和软骨细胞。通过超离心法分离BMSC-Exos。用透射电镜和Western blot鉴定外泌体。荧光显微镜下观察BMSC-Exos向软骨细胞的内化。采用MTT法、Hoechst 33324染色法和线粒体损伤检测法检测IL-1诱导的软骨细胞存活率和凋亡情况。蛋白质印迹法检测p38、ERK和Akt的磷酸化。结果显示BMSC-Exos呈圆形。将BMSC-Exos与软骨细胞共培养，观察软骨细胞对BMSC-Exos的摄取情况。IL-1作用后软骨细胞活力下降，出现凋亡，线粒体膜电位发生明显变化，而BMSC-Exos作用后上述变化基本消失。与IL-1组相比，BMSC-Exos组p38和ERK的磷酸化水平降低，Akt的磷酸化水平升高。目前的研究表明，BMSC-Exos抑制IL-1诱导的细胞凋亡，p38，ERK和Akt通路参与其中。BMSC-Exo可能代表一种新的无细胞治疗OA的方法。
Osteoarthritis (OA) is the most common chronic joint disease worldwide. Chondrocyte, as the only resident cell type in cartilage, its apoptosis is of pathogenetic significance in OA. Mesenchymal stem cell (MSC)-based-therapy has been proved effective in OA in animals and clinical studies. Nowadays, the regenerative potential of MSC-based therapy is mostly attributed to its paracrine secretion, in which exosomes may play an important role. In the present study, we aimed to find out the significance of MSC-derived exosomes (MSC-Exos) on the viability of chondrocytes under normal and inflammatory conditions. Bone marrow MSCs (BMSCs) and chondrocytes from rabbits were cultured in vitro. BMSC-Exos were isolated by an ultracentrifugation method. Transmission electron microscopy and Western blot were used to identify exosomes. The internalization of BMSC-Exos into chondrocytes was observed by fluorescent microscope. The viability and apoptosis of chondrocytes induced by IL-1 were tested through MTT method, Hoechst33324 dying, and mitochondrial damage measurement. Phosphorylation of p38, ERK, and Akt were evaluated by Western blot. The results showed that BMSC-Exos were round-shaped. Co-culturing BMSC-Exos with chondrocytes could observe the uptake of BMSC-Exos by chondrocytes. The viability decreased, apoptosis occurred, and the mitochondrial membrane potential of chondrocytes changed a lot when IL-1 were given, but all the changes were almost abolished when BMSC-Exos was added. Furthermore, the phosphorylation of p38 and ERK were inhibited, and phosphorylation of Akt was promoted by BMSC-Exos compared with IL-1 group. The present study demonstrated that BMSC-Exos inhibited mitochondrial-induced apoptosis in response to IL-1, and p38, ERK, and Akt pathways were involved. BMSC-Exo might represent a novel cell-free therapeutic approach for the treatment of OA.