微小 RNA-615-3p 通过抑制骨髓间充质干细胞的成软骨分化促进骨关节炎的进展。
MicroRNA-615-3p promotes the osteoarthritis progression by inhibiting chondrogenic differentiation of bone marrow mesenchymal stem cells.
机构信息
Department of Orthopedics, The First People's Hospital of Wujiang District, Suzhou, China.
出版信息
Eur Rev Med Pharmacol Sci. 2018 Oct;22(19):6212-6220. doi: 10.26355/eurrev_201810_16027.
OBJECTIVE
To investigate whether microRNA-615-3p participates in the development and progression of osteoarthritis by regulating chondrogenic differentiation of bone marrow mesenchymal stem cells.
MATERIALS AND METHODS
Bone marrow mesenchymal stem cells (BMSCs) were isolated from rat bone marrow and identified by flow cytometry. After chondrogenic differentiation was induced in BMSCs, expression levels of chondrogenic-specific genes were then detected by quantitate Real-time polymerase chain reaction (qRT-PCR). Expression levels of inflammatory cytokines were detected by enzyme-linked immunosorbent assay (ELISA). Protein expression of SOX9 after overexpression or knockdown of microRNA-615-3p was detected by Western blot, respectively.
RESULTS
MicroRNA-615-3p was down-regulated in the process of chondrogenic differentiation of BMSCs. The mRNA expressions of chondrogenic-specific markers, COL2A1, COL10A1, ACAN and MATN3 were decreased after microRNA-615-3p overexpression in BMSCs. Overexpressed microRNA-615-3p down-regulated protein expression of SOX9. Expression levels of inflammatory cytokines, including interleukin-1 (IL-1), interleukin-6 (IL-6), interleukin-α (IL-α) were increased after overexpression of microRNA-615-3p, while inhibition of microRNA-615-3p expression obtained the opposite result. In addition, overexpression of SOX9 rescued the effect induced by microRNA-615-3p on inflammatory cytokines.
CONCLUSIONS
MicroRNA-615-3p participates in the development and progression of osteoarthritis by increasing the expressions of inflammatory cytokines and inhibiting chondrogenic differentiation of BMSCs.
目的
通过调控骨髓间充质干细胞的软骨分化来研究微小 RNA-615-3p 是否参与骨关节炎的发生发展。
材料与方法
从大鼠骨髓中分离骨髓间充质干细胞（BMSCs），并通过流式细胞术进行鉴定。在 BMSCs 诱导软骨分化后，通过定量实时聚合酶链反应（qRT-PCR）检测软骨特异性基因的表达水平。通过酶联免疫吸附试验（ELISA）检测炎症细胞因子的表达水平。通过 Western blot 分别检测过表达或敲低微小 RNA-615-3p 后 SOX9 的蛋白表达水平。
结果
微小 RNA-615-3p 在 BMSCs 软骨分化过程中呈下调表达。在 BMSCs 中转染微小 RNA-615-3p 后，软骨特异性标志物 COL2A1、COL10A1、ACAN 和 MATN3 的 mRNA 表达降低。过表达微小 RNA-615-3p 下调 SOX9 的蛋白表达。过表达微小 RNA-615-3p 后，炎症细胞因子的表达水平，包括白细胞介素-1（IL-1）、白细胞介素-6（IL-6）和白细胞介素-α（IL-α）增加，而抑制微小 RNA-615-3p 的表达则得到相反的结果。此外，过表达 SOX9 可挽救微小 RNA-615-3p 对炎症细胞因子的作用。
结论
微小 RNA-615-3p 通过增加炎症细胞因子的表达和抑制 BMSCs 的软骨分化参与骨关节炎的发生发展。
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