[miR-877-3p通过抑制小鼠骨髓间充质干细胞MCP-1分泌和T淋巴细胞的迁移和凋亡导致小鼠骨质疏松]。

Jie Qin, Yan Zhang
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引用次数: 0

摘要

目的探讨miR-877-3p对骨间充质干细胞(BMSCs)迁移和凋亡T淋巴细胞的影响。方法建立双侧卵巢切除(OVX)假手术所致骨质疏松模型。术后8周,采用显微ct检测两组骨参数。ELISA法检测骨髓间充质干细胞单核细胞趋化蛋白1(MCP-1)水平。OVX组和sham组分别与T淋巴细胞共培养BMSC。采用PKH26染色TranswellTM法观察两组小鼠T淋巴细胞的迁移能力,流式细胞术检测T淋巴细胞的凋亡情况。采用反转录PCR检测miR-877-3p在骨髓间充质干细胞中的表达。细胞转染后miR-877-3p过表达或下调。ELISA法检测各组骨髓间充质干细胞分泌的MCP-1水平。采用上述方法检测T淋巴细胞的迁移和凋亡情况。结果OVX组骨小梁数量和骨密度均低于假手术组。OVX组骨髓间充质干细胞MCP-1分泌水平、趋化和凋亡T淋巴细胞能力均低于sham组。OVX组BMSC中miR-877-3p的表达水平高于sham组。过表达BMSC miR-877-3p后,BMSCs分泌的MCP-1水平和凋亡T淋巴细胞水平下降,而下调miR-877-3p后,结果相反。结论miR-877-3p可能通过抑制骨髓间质干细胞MCP-1分泌和T淋巴细胞的迁移和凋亡而成为骨质疏松的原因之一。
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[miR-877-3p causes osteoporosis in mice by inhibiting MCP-1 secretion from mouse bone marrow mesenchymal stem cells and the migration and apoptosis of T lymphocytes].

Objective To investigate the effects of miR-877-3p on migration and apoptotic T lymphocytes of bone mesenchymal stem cells (BMSCs). Methods The model of osteoporosis induced by bilateral ovariectomy (OVX) and sham operation was established. At 8 weeks after operation, the bone parameters of the two groups were detected by micro-CT. The levels of monocyte chemotactic protein 1(MCP-1) in BMSCs were detected by ELISA. BMSC in OVX group and sham group were co-cultured with T lymphocytes, respectively. The migration ability of T lymphocytes in the two groups was observed by TranswellTM assay with PKH26 staining and apoptosis of T lymphocytes were detected by flow cytometry. Reverse transcription PCR was used to detect the expression of miR-877-3p in BMSCs. miR-877-3p was overexpressed or down-regulated by cell transfection. The level of MCP-1 secreted by BMSCs in each group was detected by ELISA. The migration and apoptosis of T lymphocytes were detected by the above methods. Results The number of trabecular bone and bone mineral density in OVX group were lower than those in sham group. The levels of MCP-1 secretion, chemotactic and apoptotic T lymphocyte ability of BMSCs in OVX group were also lower than those in sham group. The expression level of miR-877-3p in BMSC in OVX group was higher than that in sham group. After overexpression of BMSC miR-877-3p, the levels of MCP-1 secreted from BMSCs, and apoptotic T lymphocytes decreased, while the results were opposite after down-regulation of miR-877-3p. Conclusion miR-877-3p may be one of the causes of osteoporosis by inhibiting MCP-1 secretion of BMSCs and the migration and apoptosis of T lymphocytes.

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