基于IL-33/ST2 信号通路探讨蛋白酶抑制剂硼替佐米治疗类风湿关节炎的作用机制

Effect of protease inhibitor bortezomib on treatment of rheumatoid ar⁃thritis by affecting IL-33/ST2 signaling pathway

目的:基于白细胞介素33(IL-33)/基质裂解素2(ST2)信号通路探讨蛋白酶抑制剂硼替佐米治疗类风湿关节炎(RA)的作用机制。方法:将40只Wistar大鼠随机分为4组:对照组、模型组和硼替佐米低、高剂量组,每组各10只。除对照组外,其余组大鼠构建RA模型。硼替佐米低、高剂量组分别腹腔注射0.2 mg/kg和0.5 mg/kg硼替佐米,对照组和模型组腹腔注射等量生理盐水,每天1次,连续给药21 d。观察各组大鼠一般情况,计算足跖肿胀度,并进行炎症评分;HE染色观察各组大鼠踝关节组织病理变化;全自动生化分析仪检测血红蛋白含量、血小板(PLT)总数、血清肌酐(SCr)水平和血尿素氮(BUN)水平;ELISA法检测各组大鼠血清中IL-6、肿瘤坏死因子α(TNF-α)、IL-33和ST2水平;Western blot法检测各组大鼠踝关节组织中IL-33和ST2蛋白表达水平。结果:在造模后7、14和21 d,与对照组相比,模型组大鼠足跖肿胀度显著升高(P<0.05);与模型组相比,硼替佐米低、高剂量组大鼠足跖肿胀度依次降低(P<0.05)。给药结束后,与对照组相比,模型组滑膜细胞增多,排列紊乱,关节腔中存在大量炎性渗出物,大鼠炎症评分、血PLT总数、SCr水平和BUN水平显著升高,血清中IL-6、TNF-α、IL-33和ST2水平及踝关节组织中IL-33和ST2蛋白表达水平亦显著升高(P<0.05);与模型组相比,硼替佐米低、高剂量组大鼠关节腔炎性渗出物减少,炎症评分、血PLT总数、SCr水平和BUN水平依次降低,血清中IL-6、TNF-α、IL-33和ST2水平及踝关节组织中IL-33和ST2蛋白表达水平亦依次降低(P<0.05)。结论:硼替佐米可能通过调控IL-33/ST2信号通路,减轻RA大鼠关节炎症和肿胀。

AIM:To investigate the effect of bortezomib,a protease inhibitor,on the treatment of rheumatoid arthritis(RA)and it mechanism,based on interleukin-33(IL-33)/suppression of tumorigenicity 2(ST2)signaling path⁃way.METHODS:A total of 40 Wistar rats were randomly divided into 4 groups:control group,model group,and lowand high-dose bortezomib groups,with 10 rats in each group.In addition to control group,the rats in other groups were used to construct RA model.Bortezomib was given intraperitoneally at 0.2 mg/kg and 0.5 mg/kg in low-and high-dose bortezomib groups,respectively,while the rats in control group and model group were injected with the same amount of sa⁃line,once a day for 21 d.The general situation of the rats in each group was observed,the swelling degree of the foot was calculated,and the inflammation score was evaluated.HE staining was used to observe the pathological changes of ankle joint.The automatic biochemical analyzer was used to detect blood hemoglobin content,the total number of platelets(PLT),serum creatinine(SCr)level and blood urea nitrogen(BUN)level.The serum levels of IL-6,tumor necrosis factor-α(TNF-α),IL-33 and ST2 were measured by ELISA.The protein expression of IL-33 and ST2 in ankle tissues of each group was determined by Western blot.RESULTS:On the 7th,14th and 21th days after modeling,compared with control group,the degree of paw swelling in model group was significantly increased(P<0.05).Compared with model group,the swelling degree of paw in low-and high-dose groups was decreased(P<0.05).At the end of administration,compared with control group,the synovial cells in model group were increased and in disorder,with a lot of inflammatory exudates in the articular cavity,and the inflammatory score,the levels of PLT,SCr and BUN,the serum levels of IL-6,TNF-α,IL-33 and ST2,and the protein expression of IL-33 and ST2 in ankle tissues were significantly increased(P<0.05).Compared with model group,the inflammatory exudates in the articular cavity of the rats in low-and high-dose bortezomib groups were decreased,and the inflammatory score,the levels of PLT,SCr and BUN,the serum levels of IL-6,TNF-α,IL-33 and ST2,and the protein expression of IL-33 and ST2 in ankle tissues were decreased(P<0.05).CON⁃CLUSION:Bortezomib may reduce the inflammation and swelling of the joints in RA rats by regulating the IL-33/ST2 sig⁃naling pathway.