吉林大学学报(医学版) ›› 2021, Vol. 47 ›› Issue (2): 390-396.doi: 10.13481/j.1671-587X.20210219

• 基础研究 • 上一篇    下一篇

白芍总苷介导TLR4/NF-κB信号通路对类风湿关节炎模型大鼠的治疗作用及其机制

燕丽君,佟胜全(),刘静,高冬梅,陈南芳,胡杰   

  1. 河北医科大学附属唐山工人医院风湿免疫科,河北 唐山 063000
  • 收稿日期:2020-08-30 出版日期:2021-03-28 发布日期:2021-03-25
  • 通讯作者: 佟胜全 E-mail:lijunyan84@163.com
  • 作者简介:燕丽君(1979-),女,河北省秦皇岛市人,副主任医师,医学硕士,主要从事风湿免疫学等方面的研究。
  • 基金资助:
    河北省科技厅重点研发项目(172777146)

Therapeutic effect of total glucosides of paeony in model rats with rheumatoid arthritis by mediating TLR4/NF-κB signaling pathway and its mechanisim

Lijun YAN,Shengquan TONG(),Jing LIU,Dongmei GAO,Nanfang CHEN,Jie HU   

  1. Department of Rheumatology Immunology,Tangshan Workers’ Hospital,Hebei Medical University,Hebei 063000,China
  • Received:2020-08-30 Online:2021-03-28 Published:2021-03-25
  • Contact: Shengquan TONG E-mail:lijunyan84@163.com

摘要: 目的

探讨白芍总苷(TGP)通过调控Toll样受体4(TLR4)/核因子κB(NF-κB)信号通路对类风湿关节炎(RA)大鼠关节情况、炎症因子及凋亡因子的影响,并阐明其作用机制。

方法

利用牛Ⅱ型胶原蛋白溶液和完全弗氏佐剂构建胶原诱导性关节炎(CIA)大鼠模型,将CIA模型大鼠随机分为CIA组、低剂量(25 mg·kg-1)TGP组(TGP-LD组)、中剂量(50 mg·kg-1)TGP组(TGP-MD)和高剂量(100 mg·kg-1) TGP 组(TGP-HD组),每组10只;另选取10只健康SD大鼠作为对照组。记录各组大鼠体质量、关节炎指数及关节肿胀度;采用酶联免疫吸附试验(ELISA)法检测各组大鼠血清中肿瘤坏死因子α(TNF-α), 白细胞介素6(IL-6)和白细胞介素1β(IL-1β)水平;采用实时荧光定量PCR(RT-qPCR)和Western blotting法检测各组大鼠关节滑膜组织中TLR4、NF-κB、活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved caspase-3)、B细胞淋巴瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)mRNA和蛋白表达水平。

结果

与对照组比较,CIA组大鼠体质量和关节滑膜组织中Cleaved caspase-3和Bax mRNA和蛋白表达水平明显降低(P<0.05),关节炎指数、关节肿胀度、关节滑膜组织中Bcl-2、TLR4、NF-κB p65 mRNA及蛋白表达水平,血清中TNF-α、IL-6和IL-1β水平明显升高(P<0.05)。与CIA组比较,不同剂量TGP组大鼠体质量和关节滑膜组织中Cleaved caspase-3及Bax mRNA和蛋白表达水平明显升高(P<0.05),关节炎指数、关节肿胀度、关节滑膜组织中Bcl-2、TLR4、NF-κB p65 mRNA和蛋白表达水平,血清中TNF-α、IL-6和IL-1β水平明显降低(P<0.05)。与TGP-LD组比较, TGP-MD组和TGP-HD组大鼠体质量和关节滑膜组织中Cleaved caspase-3及Bax mRNA和蛋白表达水平明显升高(P<0.05),关节炎指数、关节肿胀度、关节滑膜组织中Bcl-2、TLR4、NF-κB p65 mRNA和蛋白表达水平,血清中TNF-α、IL-6、IL-1β水平明显降低(P<0.05)。

结论

TGP可通过抑制TLR4/NF-κB信号通路,改善CIA症状,降低炎症反应,并促进关节滑膜组织细胞的凋亡。

关键词: 白芍总苷, Toll样受体4, 核因子κB, 类风湿关节炎, 炎症, 滑膜

Abstract: Objective

To investigate the effects of total glucosides of paeony (TGP) on joint condition, inflammatory factors and apoptosis factors in the rheumatoid arthritis (RA) rats by regulating the Toll-like receptor 4(TLR4) /nuclear factor-κB(NF-κB) signaling pathway, and to clarify its mechanism.

Methods

The collagen-induced arthritis (CIA) rat models were established with bovine type Ⅱ collagen and complete Freund’s adjuvant, and the CIA model rats were randomly divided into CIA group, low dose (25 mg·kg-1)of TGP(TGP-LD) group, middle dose (50 mg·kg-1) of TGP(TGP-MD) group, and high dose (100 mg·kg-1) of TGP(TGP-HD), with 10 rats in each group.Another 10 healthy SD rats were selected as control group . The body weights, arthritis indexes and joint swelling degrees of the rats in each group were measured, and the levels of tumor necrosis factor-α (TNF-α),interleukin-6 (IL-6) and interleukin-1β (IL-1β) in serum of the rats in each group were detected by ELISA method. The mRNA and protein expression levels of TLR4, NF-κB, Cleaved caspase-3, B cell lymphoma -2 (Bcl-2) and Bcl-2 associated X protein (Bax) in joint synovium tissue of the rats in each group were detected by Real-time fluorescence quantitative PCR(RT-qPCR) and Western blotting methods.

Results

Compared with control group, the body weight and the expression levels of Cleaved caspase-3 and Bax mRNA and proteins in joint synovium tissue of the rats in CIA group were significantly decreased (P<0.05), while the arthritis index, the joint swelling degree, the expression levels of Bcl-2, TLR4,and NF-κB p65 mRNA and proteins in joint synovium tissue, and the TNF-α, IL-6, and IL-1β levels in serum of the rats in CIA group were significantly increased (P<0.05). Compared with CIA group,the body weights and the expression levels of Cleaved caspase-3 and Bax mRNA and protein in joint synovium tissue of the rats in different doses of TGP groups were significantly increased(P<0.05), while the arthritis indexes,the joint swelling degrees, the expression levels of Bcl-2, TLR4,and NF-κB p65 mRNA and proteins in joint synovium tissue, and the TNF-α, IL-6, IL-1β levels in serum of the rats were significantly decreased(P<0.05).Compared with TGP-LD group,the body weights and the expression levels of Cleaved caspase-3 and Bax mRNA and proteins in joint synovium tissue of the rats in TGP-MD group and TGP-HD group were significantly increased(P<0.05),while the arthritis indexes, the joint swelling degrees, the expression levels of TLR4,NF-κB,p65 and Bcl-2 in joint synovium tissue, and the TNF-α, IL-6, and IL-1β levels in serum of the rats were significantly decreased(P<0.05).

Conclusion

TGP can improve the symptoms of CIA, reduce the inflammatory reaction and promote the apoptosis of synovial tissue by inhibiting TLR4/NF- κB signal pathway.

Key words: total glucoside of paeony, Toll-like receptor 4, nuclear factor-κB, rheumatoid arthritis, inflammation, synovial

中图分类号: 

  • R684.3