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

[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].

作者信息

Qin Jie, Zhang Yan

机构信息

Stomatological Hospital of Chongqing Medical University, Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences, Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Chongqing 401147, China.

Stomatological Hospital of Chongqing Medical University, Chongqing Key Laboratory of Oral Diseases and Biomedical Sciences, Chongqing Municipal Key Laboratory of Oral Biomedical Engineering of Higher Education, Chongqing 401147, China. *Corresponding author, E-mail:

出版信息

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2023 Jun;39(6):481-487.

Abstract

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 Transwell 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.

摘要

目的 探讨miR-877-3p对骨间充质干细胞(BMSCs)迁移及T淋巴细胞凋亡的影响。方法 建立双侧卵巢切除(OVX)诱导的骨质疏松模型及假手术模型。术后8周,通过显微CT检测两组的骨参数。采用酶联免疫吸附测定(ELISA)法检测BMSCs中单核细胞趋化蛋白1(MCP-1)的水平。分别将OVX组和假手术组的BMSC与T淋巴细胞共培养。采用PKH26染色的Transwell实验观察两组T淋巴细胞的迁移能力,通过流式细胞术检测T淋巴细胞的凋亡情况。采用逆转录聚合酶链反应(RT-PCR)检测BMSCs中miR-877-3p的表达。通过细胞转染使miR-877-3p过表达或下调。采用ELISA法检测各组BMSCs分泌的MCP-1水平。通过上述方法检测T淋巴细胞的迁移和凋亡情况。结果 OVX组的骨小梁数量和骨密度低于假手术组。OVX组BMSCs的MCP-1分泌水平、趋化及凋亡T淋巴细胞的能力也低于假手术组。OVX组BMSC中miR-877-3p的表达水平高于假手术组。BMSC中miR-877-3p过表达后,BMSCs分泌的MCP-1水平及凋亡T淋巴细胞数量降低,而miR-877-3p下调后结果相反。结论 miR-877-3p可能通过抑制BMSCs分泌MCP-1以及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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