骨髓间充质干细胞来源的外泌体抑制地塞米松诱导的C2C12肌管萎缩  

Bone marrow mesenchymal stem cell-derived exosomes inhibit dexamethasone-induced C2C12 myotube atrophy

作　　者：柯义兵 丁永宏 阿布都克热木·达吾提 郭浩然 兰志杰 王勇平[3] KE Yi-bing;DING Yong-hong;A budoukeremu·DA-wuti;GUO Hao-ran;LAN Zhi-jie;WANG Yong-ping(The First Clinical Medical College of Lanzhou University,Lanzhou 730000,China;Ningxia Medical University,Yinchuan 750000,China;Dept of Orthopaedics,First Hospital of Lanzhou University,Lanzhou 730000,China)

机构地区：[1]兰州大学第一临床医学院,甘肃兰州730000 [2]宁夏医科大学,宁夏银川750000 [3]兰州大学第一医院骨科,甘肃兰州730000

出　　处：《中国药理学通报》2025年第1期50-56,共7页Chinese Pharmacological Bulletin

基　　金：甘肃省高等学校科研项目(No 2023CYZC-03)。

摘　　要：目的研究骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)来源的外泌体(exsomes,EXOs)对地塞米松(dexamethasone,DEX)诱导的C2C12肌管萎缩的影响。方法(1)分离提取培养C57BL/6J小鼠BMSCs。(2)提取并鉴定BMSCs-EXOs。(3)C2C12细胞成肌分化。(4)将肌管细胞分为对照组(Control,2%马血清培养基培养48 h)、地塞米松组(DEX、浓度为10μmol·L^(-1)的DEX干预肌管48 h)、外泌体组(EXOs、外泌体干预肌管48 h)、外泌体抑制剂组(GW4869、10μmol的GW4869干预BMSCs后提取的外泌体干预肌管48 h)。48 h后测量各组肌管直径,CCK-8法检测各组细胞活力,Western blot检测各组肌肉萎缩及成肌分化相关蛋白的表达量。结果BMSCs呈长梭形,BMSCs-EXOs在透射电镜下为圆形的双层膜结构,直径约200 nm,且高表达CD9、CD63和CD81。相比于Control组,DEX组细胞活性降低(P<0.01),肌管细胞直径减少(P<0.01),atrogin-1(P<0.05)和MuRF-1(P<0.01)的表达明显上调,MYOD(P<0.01)蛋白表达降低;相比于DEX组,BMSCs-EXOs组细胞活性升高(P<0.01),肌管细胞直径增加(P<0.01),atrogin-1(P<0.05)和MuRF-1(P<0.01)的表达明显下调,MYOD(P<0.01)蛋白表达升高;相比于BMSCs-EXOs组,BMSCs-EXOs(GW4869)组细胞活性降低(P<0.05),肌管细胞直径减少(P<0.01),atrogin-1(P<0.05)和MuRF-1(P<0.05)的表达上调,MYOD(P<0.01)蛋白表达降低。结论骨髓间充质干细胞来源的外泌体(BMSCs-EXOs)可抑制地塞米松诱导的C2C12肌管萎缩。Aim To investigate the effect of exosomes derived from bone marrod-derived mesenchymal stem cells(BMSCs)on dexamethasone-induced C2C12 muscular canal atrophy.Methods(1)C57BL/6J mouse bone marrow mesenchymal stem cells were isolated and cultured by whole bone marrow adhesion method.(2)Extraction and identification of BMSCs EXOs were performed.(3)Myogenic differentiation of C2C12 cells was carried out.(4)The successfully differentiated myotubes were divided into the control group(cultured in 2%equine serum medium for 48 h),dexamethasone group(dexamethasone,DEX,10μmol·L^(-1) concentration of DEX interfered with myotubes for 48 h),and exosomes group(exosomes,EXOs,interfered with myotubes for 48 h),exosome inhibitor group(exosomes extracted from BMSCs after 10μm GW4869 intervention,interfered with myotubes for 48 h).48 h later,the morphology and diameter of muscle tubes were observed and measured by microscope.Cell viability of each group was detected by CCK-8 method.The expression levels of atrogin-1 and MuRF-1,myogenic differentiation antigen(MYOD)in each group were detected by Western blot.Results BMSCs were long spusiform,and BMSCS-EXOS showed a circular bilayer structure under transmission electron microscopy,with a diameter of about 200 nm.CD9,CD63 and CD81 were highly expressed.Compared with the control group,cell activity in DEX group decreased(P<0.01),diameter of myotubes decreased(P<0.01),expressions of atrogin-1(P<0.05)and MuRF-1(P<0.01)were significantly up-regulated,and expression of MYOD(P<0.01)was significantly down-regulated.Compared with the DEX group,cell activity in the BMSCs-EXOs group increased(P<0.01),diameter of myotubes increased(P<0.01),expressions of atrogin-1(P<0.05)and MuRF-1(P<0.01)were significantly down-regulated,and expression of MYOD(P<0.01)was up-regulated.Compared with the BMSCs-EXOs group,cell activity of the BMSCs-EXOs(GW4869)group decreased(P<0.05),diameter of myotubes decreased(P<0.01),expressions of atrogin-1(P<0.05)and MuRF-1(P<0.05)were up-regulated,and expression of MYOD(P

关 键 词：骨髓间充质干细胞 外泌体 地塞米松 肌管 肌肉萎缩 C2C12 

分 类 号：R965[医药卫生—药理学]