卡非佐米通过抑制FLS增殖对大鼠类风湿关节炎模型的作用研究

The effect of carfilzomib on rheumatoid arthritis in rats through inhibiting proliferation of FLS

目的本研究通过第二代蛋白酶体抑制剂卡非佐米(carfilzomib,CFZ)对大鼠佐剂诱导型关节炎(adjuvant induced arthritis,AIA)的作用,研究第二代蛋白酶体抑制剂卡非佐米(carfilzomib,CFZ)对大鼠类风湿关节炎(rheu matoid arthritis,RA)的治疗作用及其机制。方法通过注射弗氏佐剂诱导SD大鼠建立AIA大鼠模型。试验分组:低剂量组(1.0mg/kg)、中剂量组(1.5mg/kg)、高剂量组(2.0mg/kg)、预治疗组(1.5mg/kg)、同时治疗组(1.5mg/kg)、模型对照组及空白对照组。观察各组大鼠后足关节情况;TRAP染色研究各组破骨细胞抑制水平;采用ELISA分析外周血IL-1β、IL-6、TNF-α和IFN-γ表达水平;采用CCK-8分析不同剂量治疗组对纤维母细胞样滑膜细胞(fibroblast-like synoviocytes,FLS)的抑制水平。结果足趾观察结果显示各治疗组大鼠后足病变情况均优于模型对照组;TRAP染色结果显示,与AIA模型对照组相比,各CFZ治疗组的破骨细胞均明显受到抑制;ELISA分析显示各CFZ治疗组外周血中IL-1β、IL-6、TNF-α和IFN-γ表达显著降低(P<0.01);CCK-8结果显示3种剂量治疗组均显著抑制FLS增生(P<0.01)。结论CZF通过抑制AIA大鼠FLS增殖,降低IL-1β、IL-6、TNF-α和IFN-γ表达,调控炎症反应,达到治疗大鼠AIA的目的,CFZ是一种具有治疗类风湿关节炎潜力的药物。

Objective To observe the action of carfilzomib ( CFZ) in adjuvant-induced arthritis ( AIA) rat model,and explore the therapeutic effect and mechanism of carfilzomib on rheumatoid arthritis ( RA) . Methods Freund’s adjuvants were used to induce arthritis in rats,and AIA models were successfully constructed. All rats were divided into seven groups: low-dose treatment group ( 1. 0 mg /kg) ,middle-dose treatment group ( 1. 5 mg /kg) ,high-dose treatment group ( 2. 0 mg /kg) ,pre-treatment group ( 1. 5 mg /kg) ,concurrent treatment group ( 1. 5 mg /kg) ,AIA model control group and normal control group. The changes of hind feet of rats were observed and recorded. TRAP staining was used to analyses the inhabitation level of osteoclasts in different groups. ELISA was used to analyze the expression level of IL-1β, IL-6,TNF-α,and IFN-γ in peripheral blood. The CCK-8 assay was used to detect the proliferation activity of fibroblastlike synoviocytes. Results The observation on the changes of hind feet showed that the rats in all treatment group had the better joint condition than AIA model group. TRAP staining results showed that constrain of osteoclasts in the treatment group was much higher than the AIA model group. ELISA showed that level of IL-1β,IL-6,TNF-α,and IFN-γ in peripheral blood of all 5 treatment group were much lower those in AIA model group. CCK-8 results showed that lowdose, middle-dose and high-dose of carfilzomib had inhibited the proliferation activity of fibroblast-like synoviocytes significantly. Conclusion Carfilzomib is effective for the adjuvant-induced arthritis rats through suppressing the proliferation activity of fibroblast-like synoviocytes,down-regulation the expression of IL-1β,IL-6,TNF-α and IFN-γ to regulate inflammatory response,has a potential therapeutic value for RA.