骨髓间充质干细胞源外泌体对软骨细胞迁移、增殖和凋亡的影响

Effects of Bone Marrow Mesenchymal Stem Cell-derived Exosome on the Migration,Proliferation and Apoptosis of Chondrocytes

目的探讨大鼠骨髓间充质干细胞(BMSC)分泌的外泌体(BMSC-Exo)对原代软骨细胞迁移、增殖和凋亡的影响。方法超速离心结合超滤浓缩法提取BMSC-Exo,通过透射电镜和Western blot法鉴定外泌体。分别使用浓度为25、50和100μg/ml的BMSC-Exo处理软骨细胞,划痕实验检测软骨细胞的迁移,CCK-8法检测软骨细胞的增殖,流式细胞术检测软骨细胞的凋亡。为模拟骨关节炎(OA)病理状态,同时设置IL-1β(10ng/ml)对照组。结果与细胞培养基对照组比较,BMSC-Exo作用使软骨细胞的划痕愈合率显著升高(12h,P<0.05;24h,P<0.01),细胞增殖速度明显加快(P<0.01),细胞凋亡比例显著降低(P<0.01),且均存在时间和剂量依赖性。IL-1β刺激可抑制软骨细胞的迁移和增殖,促进软骨细胞的凋亡,而BMSC-Exo可显著削减IL-1β的作用效果。结论BMSC-Exo可促进软骨细胞的迁移运动和增殖能力,抑制软骨细胞发生凋亡,可对软骨细胞的生长发挥重要的调控作用。

Objective To investigate the effects of rat bone marrow mesenchymal stem cell-derived exosome(BMSC-Exo)on the migration,proliferation and apoptosis of chondrocytes.Methods BMSC-Exo were isolated and identified by transmission electron microscope and Western blot.Primary chondrocytes were treated with different concentrations of BMSC-Exo(25,50 and 100μg/ml),and the cell scratch assay,CCK-8 assay and flow cytomertry method were used to assess the migration,proliferation and apoptosis of chondrocytes respectively.To simulate the pathological status of osteoarthritis(OA),the IL-1βcontrol groups were set at the same time.Results Compared with control groups,the scratch healing and cell proliferation ability were significantly increased,the apoptotic rates were obviously reduced of cells in BMSC-Exo treated groups.IL-1βstimulation could inhibit the migration and proliferation of chondrocytes,promote the apoptosis of chondrocytes,while BMSC-Exo could significantly reduce the effects of IL-1β.Conclusion BMSC-Exo could promote the migration and proliferation of chondrocytes,and inhibit the apoptosis of chondrocytes,such play an important role in the growth regulation of chondrocytes.