1,25-二羟维生素D3通过调控类风湿关节炎大鼠模型中的微小RNA-22对成纤维样滑膜细胞增殖及促炎细胞因子表达的影响
Effect of 1,25-(OH)2D3 on Proliferation of Fibroblast-Like Synoviocytes and Expressions of Pro-Inflammatory Cytokines through Regulating MicroRNA-22 in a Rat Model of Rheumatoid Arthritis.
作者信息
Fan Ping, He Lan, Hu Nan, Luo Jing, Zhang Jing, Mo Ling-Fei, Wang Yan-Hua, Pu Dan, Lv Xiao-Hong, Hao Zhi-Ming, Ding Chang-Hai, Xue Wu-Jun, Li Yang
机构信息
Department of Rheumatism and Immunology, Xi'an, China.
Department of Renal Transplantation, Center of Nephrology, the First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, China.
出版信息
Cell Physiol Biochem. 2017;42(1):145-155. doi: 10.1159/000477123. Epub 2017 May 12.
OBJECTIVE
This study aims to investigate the regulatory mechanism of 1,25-(OH)2D3 on the proliferation of fibroblast-like synoviocytes (FLS) and expressions of pro-inflammatory cytokines in rheumatoid arthritis (RA) rats via microRNA-22 (miR-22).
METHODS
A rat model of RA was established with a subcutaneous injection of type II collagen. After treated with different concentrations of 1,25-(OH)2D3 the proliferation of FLS was estimated by the MTT method, and the optimal concentration of 1,25-(OH)2D3 was selected for further experiments. Cell proliferation was detected by MTT. Cell cycle and apoptosis were analyzed by FCM. The IL-1β, IL-6, IL-8, and PGE2 protein expressions were determined by ELISA, and MMP-3, INOS, and Cox-2 mRNA expressions were measured by qRT-PCR.
RESULTS
The rat model of RA was successfully established. Compared with the blank group, the 1,25-(OH)2D3 and miR-22 inhibitors groups exhibited higher proliferation inhibition and apoptosis rates, lower levels of pro-inflammatory cytokines (IL-1β, IL-6, IL-8, and PGE2), and decreased mRNA expressions of MMP-3, INOS, and Cox-2. The miR-22 mimics group had lower proliferation inhibition and apoptosis rates, elevated expressions of pro-inflammatory cytokines and MMP-3, INOS, and Cox-2 than the blank group. In contrast to the 1,25-(OH)2D3 group, the proliferation inhibition and apoptosis rates were down-regulated, and the expressions of pro-inflammatory cytokines and MMP-3, INOS, and Cox-2 were up-regulated in the 1,25-(OH)2D3 + miR-22 mimics group.
CONCLUSION
Our study demonstrated that 1,25-(OH)2D3 inhibits the proliferation of FLS and alleviates inflammatory response in RA rats by down-regulating miR-22.
目的
本研究旨在通过微小RNA-22（miR-22）探讨1,25-（OH）₂D₃对类风湿关节炎（RA）大鼠成纤维样滑膜细胞（FLS）增殖及促炎细胞因子表达的调控机制。
方法
采用皮下注射Ⅱ型胶原建立RA大鼠模型。用不同浓度的1,25-（OH）₂D₃处理后，通过MTT法评估FLS的增殖情况，并选择1,25-（OH）₂D₃的最佳浓度进行进一步实验。用MTT检测细胞增殖。通过流式细胞术分析细胞周期和凋亡。用ELISA法测定IL-1β、IL-6、IL-8和PGE₂蛋白表达，用qRT-PCR法检测MMP-3、INOS和Cox-2 mRNA表达。
结果
成功建立RA大鼠模型。与空白组相比，1,25-（OH）₂D₃组和miR-22抑制剂组表现出更高的增殖抑制率和凋亡率、更低水平的促炎细胞因子（IL-1β、IL-6、IL-8和PGE₂）以及MMP-3、INOS和Cox-2 mRNA表达降低。miR-22模拟物组的增殖抑制率和凋亡率低于空白组，促炎细胞因子以及MMP-3、INOS和Cox-2的表达升高。与1,25-（OH）₂D₃组相比，1,25-（OH）₂D₃ + miR-22模拟物组的增殖抑制率和凋亡率下调，促炎细胞因子以及MMP-3、INOS和Cox-2的表达上调。
结论
我们的研究表明，1,25-（OH）₂D₃通过下调miR-22抑制RA大鼠FLS的增殖并减轻炎症反应。
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