吉林大学学报(医学版) ›› 2021, Vol. 47 ›› Issue (2): 460-468.doi: 10.13481/j.1671-587X.20210227

• 临床研究 • 上一篇    下一篇

miR-26a-5p通过JAK2/STAT3 信号通路对人类风湿关节炎成纤维样滑膜细胞凋亡的影响

吴洁1,杨学华2,马玲1,范文强1,付冬冬1,高晓1,左淑飞1,梁舒1,秦艺璐1,王培山3(),郭金燕4   

  1. 1.河南省新乡市中心医院风湿免疫科,河南 新乡 453000
    2.河南省新乡市第二人民医院肾病 风湿科,河南 新乡 453000
    3.河南省新乡市中心医院门诊手术部,河南 新乡 453000
    4.郑州大学第一附属医院风湿免疫科,河南 郑州 450000
  • 收稿日期:2020-05-22 出版日期:2021-03-28 发布日期:2021-03-25
  • 通讯作者: 王培山 E-mail:ccff-218000@163.com
  • 作者简介:吴 洁(1984-),女,河北省辛集市人,主治医师,医学硕士,主要从事类风湿关节炎基础和临床方面的 研究。
  • 基金资助:
    河南省科技厅科技攻关计划项目(202102310383)

Effect of miR-26a-5p on apoptosis of human rheumatoid arthritis fibroblast-like synovial cells through JAK2/STAT3 signaling pathway

Jie WU1,Xuehua YANG2,Ling MA1,Wenqiang FAN1,Dongdong FU1,Xiao GAO1,Shufei ZUO1,Shu LIANG1,Yilu QIN1,Peishan WANG3(),Jinyan GUO4   

  1. 1.Department of Rheumatology and Immunology,Xinxiang Central Hospital,Henan Province,Xinxiang 453000,China
    2.Department of Nephrology and Rheumatology,Second People’s Hospital of Xinxiang,Henan Province,Xinxiang 453000,China
    3.Department of Outpatient Operation,Xinxiang Central Hospital,Henan Province,Xinxiang 453000,China
    4.Department of Rheumatology and Immunology,First Affiliated Hospital,Zhengzhou University,Henan Province,Zhengzhou 450000,China
  • Received:2020-05-22 Online:2021-03-28 Published:2021-03-25
  • Contact: Peishan WANG E-mail:ccff-218000@163.com

摘要: 目的

探讨miR-26a-5p对人类风湿关节炎(RA)-成纤维样滑膜细胞(FLSs)凋亡及炎性因子分泌的影响,阐明其作用机制。

方法

分离RA患者的膝关节滑膜组织RA-FLSs,将miR-26a-5p模拟物(mimic)、抑制物(inhibitor)及阴性对照转染 RA-FLSs,采用实时荧光定量聚合酶链式反应(qRT-PCR)法检测转染效果,酶联免疫吸附试验(ELISA)法检测细胞培养液中白细胞介素1β(IL-1β)、白细胞介素6(IL-6)和肿瘤坏死因子α(TNF-α)水平,CCK-8 法检测各组细胞增殖活性,AnnexinⅤ-FITC/PI与Hoechst 33342染色检测各组细胞凋亡情况,Western blotting法检测各组细胞中凋亡相关蛋白B细胞淋巴瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)与酪氨酸激酶2(JAK2)/信号传导及转录活化因子3(STAT3)信号通路相关蛋白表达水平。

结果

与空白对照组和NC-mimic组比较, miR-26a-5p mimic组RA-FLSs中 miR-26a-5p mRNA表达水平升高(P<0.05),RA-FLSs增殖活性降低(P<0.05),细胞凋亡率升高(P<0.05),细胞中Bcl-2、JAK2、p-JAK2、STAT3和p-STAT3蛋白表达水平降低(P<0.05),Bax蛋白表达水平升高(P<0.05);与空白对照组和NC-inhibitor组比较,miR-26a-5p inhibitor组RA-FLSs中 miR-26a-5p mRNA表达水平降低(P<0.05),RA-FLSs细胞增殖活性升高(P<0.05),细胞凋亡率降低(P<0.05),Bcl-2、JAK2、p-JAK2、STAT3和p-STAT3蛋白表达水平升高(P<0.05),Bax蛋白表达水平降低(P<0.05)。

结论

过表达miR-26a-5p能够抑制RA-FLSs细胞增殖,促进细胞凋亡,其作用机制可能与抑制 JAK2/STAT3信号通路的激活有关。

关键词: miR-26a-5p, 类风湿关节炎, 成纤维样滑膜细胞, 细胞凋亡, 炎性因子

Abstract: Objective

To explore the effects of miR-26a-5p on the apoptosis and secretion of inflammatory factors in the rheumatoid arthritis (RA)-fibroblast-like synovial cells (FLSs), and to clarify its mechanism of action.

Methods

The RA-FLSs from knee joint synovial tissue of patients with RA were isolated and cultured,and miR-26a-5p mimics, inhibitors and negative controls were transfected into the RA-FLSs.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR) was used to detect the effect of transfection, ELISA was used to detect the levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) in cell culture fluid, CCK-8 method was used to detect the cell proliferation activity, AnnexinⅤ-FITC/PI and Hoechst 33342 staining were used to observe the apoptosis, and Western blotting method was used to detect the expression levels of apoptosis-related proteins B cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax) and Janus kinase 2(JAK2)/signal transducer and activator of transcription(STAT3) signaling pathway-related proteins in the cells.

Results

Compared with blank control group and NC-mimic group, the expression level of miR-26a-5p in RA-FLSs in miR-26a-5p mimic group was increased (P<0.05), the proliferation activity of RA-FLSs was decreased(P<0.05),the apoptotic rate of cells was increased (P<0.05),the expression levels of Bcl-2, JAK2, p-JAK2 and STAT3 proteins were decreased(P<0.05), and the expression level of Bax protein was increased (P<0.05).Compared with blank control group and NC-inhibitor group, the expression level of miR-26a-5p in RA-FLSs in miR-26a-5p inhibitor group was decreased (P<0.05), the proliferation activity of RA-FLSs was increased (P<0.05), the apoptotic rate was decreased (P<0.05), the expression levels of Bcl-2,JAK2, p-JAK2 and STAT3 proteins were increased (P<0.05), and the expression level of Bax protein was decreased (P<0.05).

Conclusion

Overexpression of miR-26a-5p can inhibit the proliferation of RA-FLSs and promote apoptosis. Its mechanism may be related to the inhibition of the activation of JAK2/STAT3 signaling pathway.

Key words: miR-26a-5p, rheumatoid arthritis, fibroblast-like synoviocyte, apoptosis, inflammatory factor

中图分类号: 

  • R593.22