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目的探讨甲氨蝶呤通过抑制Toll样受体2(TLR2)-核因子最终激活核因子-κB(NF-κB)信号通路减轻类风湿关节炎大鼠滑膜炎的作用。方法 SD大鼠120只,根据体质量随机分成6组:对照组、模型组、雷公藤多苷片组、甲氨蝶呤组各剂量组;模型组、雷公藤多苷片组、甲氨蝶呤组各剂量组构建类风湿关节炎滑膜炎模型,并于造模成功第1天开始给予相应药物灌胃,持续给予3个月。对照组和模型组给予等体积生理盐水。试验结束后,采用Hargreaves法测定机械痛阈(PWT)及热刺激潜伏期(PWTL),RT-PCR法测定TLR2、NF-кB mRNA水平,酶联免疫吸附(ELISA)法测定TLR2、NF-кB、白细胞介素2、6(IL-2、IL-6)、肿瘤坏死因子(TNF-α)蛋白表达水平。结果与对照组比较,甲氨蝶呤各剂量PWPT、PEL评分降低,关节炎症积分升高(P <0.05);与模型组比较,甲氨蝶呤各剂量PWPT、PEL评分升高,关节炎症积分降低(P <0.05),且随着甲氨蝶呤组给药剂量的增加,PWPT、PEL评分逐渐升高,关节炎症积分逐渐降低剂量-效应关系明显(P <0.05);与雷公藤多苷片组相比,甲氨蝶呤组低、中剂量组PWPT、PEL评分降低,关节炎症积分升高;高剂量组PWPT、PEL评分略微降低,关节炎症积分略微升高(P>0.05)。与对照组比较,甲氨蝶呤组各剂量组TLR2、NF-кB mRNA、蛋白、IL-2、IL-6、TNF-α蛋白表达水平明显升高(P <0.05);与模型组比较,甲氨蝶呤组各剂量组TLR2、NF-кB mRNA及蛋白,IL-2、IL-6、TNF-α蛋白表达水平明显降低(P <0.05),且随着甲氨蝶呤组给药剂量的增加,TLR2、NF-кB mRNA及蛋白,IL-2、IL-6、TNF-α蛋白表达水平逐渐降低,剂量-效应关系明显(P <0.05);与雷公藤多苷片组相比,甲氨蝶呤组组低、中剂量组TLR2、NF-кB mRNA及蛋白,IL-2、IL-6、TNF-α蛋白表达水平明显升高;高剂量组TLR2、NF-кB mRNA及蛋白,IL-2、IL-6、TNF-α蛋白表达水平略微升高(P>0.05)。结论甲氨蝶呤能对类风湿关节炎滑膜炎具有一定程度的修复作用,其机制与甲氨蝶呤能通过抑制炎症信号通路TLR2、NF-кB mRNA、蛋白表达水平的表达,进而抑制炎症因子IL-2、IL-6、TNF-α的表达有关。
Abstract：Objective To investigate the role of methotrexate in alleviating rheumatoid arthritis in rats with rheumatoid arthritis by inhibiting the activation of nuclear factor-κB(NF-κB)signaling pathway by Tolllike receptor 2(TLR2)-nuclear factor.Methods One hundred and twenty SD rats were randomly divided into 6groups according to body weight:control group,model group,tripterygium glycosides group and 3doses groups of methotrexate.Except for the contral group,the rheumatoid arthritis synovitis model was set up in every group,and the corresponding drugs were administered on the first day of modeling,and continued for 3months.By the end of the experiment,the mechanical pain threshold(PWT)and thermal stimulation latency(PWTL)were determined by Hargreaves method,TLR2 and NF-кB mRNA levels were measured by RT-PCR,and the protein expression of TLR2,NF-кB,interleukin IL-2,IL-6and tumor necrosis factor(TNF-α)were detected by Elisa assay.Results The scores of PWT and PWTL ranking in a descending order was the control group,tripterygium glycosides group,methotrexate dose groups and model group with statistic differences(P <0.05).The arthritis scores ranking in an ascending order was the control group,tripterygium glycosides group,methotrexate dose groups and model group with statistic differences(P <0.05).The mRNA levels of TLR2,and NF-кB as well as the protein levels of TLR2,NF-кB,IL-2,IL-6,and TNF-αranking in an ascending order was the control group,tripterygium glycosides group,methotrexate dose groups and model group with statistic differences(P <0.05).The effect of the methotrexate had a dose-dependent manner(P <0.05).Moreover,the differences of the above parameters between the tripterygium glycosides group and high-dose methotrexate group were no statistical(P >0.05).Conclusion Alleviation of methotrexate on synovitis in rheumatoid arthritis rats may be related to the down-regulation of mRNA and protein levels of TLR2,and NF-кB as well as the protein levels of IL-2,IL-6and TNF-α.
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基本信息:
中图分类号:R965
引用信息:
[1]崔毅佳,王淑梅,金志国.甲氨蝶呤通过抑制TLR2-NF-κB信号通路减轻类风湿关节炎大鼠滑膜炎的作用研究[J].新疆医科大学学报,2019,42(02):211-216.
基金信息:
国家自然青年科学基金(81503135)
2018-10-22
2018
2018-12-26
2018
1
2019-02-15
2019-02-15