Department of Orthopedic, Liuzhou Worker's Hospital, Liuzhou, China.
Eur Rev Med Pharmacol Sci. 2019 Feb;23(3):1214-1220. doi: 10.26355/eurrev_201902_17014.
The aim of this study was to investigate whether bone marrow stem cells (MSCs) derived exosomes in rats could promote osteoblast proliferation and improve osteoporosis via inhibiting cell apoptosis.
MSCs in rats were isolated and cultured, followed by the identification of surface antigens via flow cytometry. The differentiation of MSCs was detected by alizarin red staining and oil red staining. After extraction from MSCs by ultracentrifugation, the size distribution of exosomes was detected by tunable resistive pulse sensing (TRPS). Specific antigens in MSCs-derived exosomes were determined by flow cytometry. Furthermore, the proliferation and viability of hFOB1.19 cells treated with MSCs-derived exosomes were detected by cell count kit-8 (CCK-8) assay. The effect of MSCs-derived exosomes on cell apoptosis was evaluated by flow cytometry. Protein expression levels of apoptosis-related genes in hFOB1.19 cells were detected by Western blot.
MSCs differentiated into osteoblasts and lipoblasts under different treatments. Meanwhile, MSCs-derived exosomes exhibited typical elongated morphology after isolation and culture for 1 and 3 days, respectively. Functionally, MSCs-derived exosomes could promote the viability of hFOB1.19 cells, and significantly increase the expression level of GLUT3. In addition, MSCs-derived exosomes remarkably downregulated apoptosis-related genes and decreased apoptosis in hFOB1.19 cells.
MSCs-derived exosomes could promote osteoblast proliferation via inhibiting cell apoptosis, eventually improving osteoporosis.
本研究旨在探讨大鼠骨髓间充质干细胞（MSCs）来源的外泌体是否能通过抑制细胞凋亡来促进成骨细胞增殖和改善骨质疏松症。
分离并培养大鼠 MSCs，通过流式细胞术鉴定其表面抗原。通过茜素红染色和油红染色检测 MSCs 的分化情况。通过超速离心从 MSCs 中提取外泌体后，使用可调电阻脉冲感应（TRPS）检测其粒径分布。通过流式细胞术确定 MSCs 来源外泌体中的特异性抗原。进一步通过细胞计数试剂盒-8（CCK-8）检测 hFOB1.19 细胞中经 MSCs 来源外泌体处理后的增殖和活力情况。通过流式细胞术评估 MSCs 来源外泌体对细胞凋亡的影响。通过 Western blot 检测 hFOB1.19 细胞中与凋亡相关的基因的蛋白表达水平。
在不同处理下，MSCs 分化为成骨细胞和脂肪细胞。同时，MSCs 来源的外泌体在分离和培养 1 天和 3 天后分别呈现出典型的细长形态。功能上，MSCs 来源的外泌体可以促进 hFOB1.19 细胞的活力，并显著增加 GLUT3 的表达水平。此外，MSCs 来源的外泌体显著下调了 hFOB1.19 细胞中与凋亡相关的基因，并减少了细胞凋亡。
MSCs 来源的外泌体可以通过抑制细胞凋亡来促进成骨细胞增殖，从而改善骨质疏松症。
