阿巴西普对类风湿关节炎慢性炎症和骨质侵蚀的作用研究

Effect of Abatacept on chronic inflammation and bone erosion in rheumatoid arthritis

目的通过建立胶原诱导关节炎(CIA)小鼠模型观察阿巴西普对类风湿关节炎(RA)慢性炎症和骨质侵蚀的影响。方法将60只BALB/c雌性小鼠按简单随机分组法分为正常组、CIA模型组、甲氨蝶呤(MTX)阳性对照组和阿巴西普低、中、高剂量组,每组各10只。除正常组外对其余5组小鼠通过胶原诱导建立CIA模型。CIA造模完成后,按成人常规临床剂量换算小鼠等效剂量给药。MTX阳性对照组灌胃给药MTX片,1次/周。阿巴西普低、中、高剂量组分别腹腔注射阿巴西普注射液25、50、100 mg/kg,200μL,1次/周。正常组和CIA模型组腹腔注射0.9%氯化钠溶液,200μL,1次/周。比较各组小鼠关节炎指数评分;采用HE染色法观察踝关节滑膜组织形态和炎症细胞浸润程度;采用免疫组化法检测踝关节TNF-α、骨桥蛋白(OPN)、NF-κB和NF-κB受体活化因子配体(RANKL)表达水平;采用ELISA法检测各组血清TNF-α、IL-6及基质金属蛋白酶(MMP)水平。结果与正常组比较,造模后,各组小鼠踝关节肿胀,活动度严重受限,关节炎指数评分升高;踝关节可见滑膜增生和明显的炎性浸润,TNF-α、OPN、NF-κB和RANKL表达均升高;血清IL-6、TNF-α、MMP-2、MMP-9水平均升高(均P<0.01)。与CIA模型组相比,MTX阳性对照组和阿巴西普中、高剂量组小鼠踝关节肿胀减轻,关节炎指数评分均降低(均P<0.05);踝关节滑膜增生和炎症浸润均明显减少,TNF-α、OPN、NF-κB、RANKL表达均降低;血清IL-6、TNF-α、MMP-2、MMP-9表达水平均降低(均P<0.01)。结论阿巴西普可以显著减轻CIA小鼠关节炎症反应,抑制滑膜增生和骨破坏。

Objective To observe the effects of Abatacept on chronic inflammation and bone erosion in rheumatoid arthritis(RA)via a mouse model of collagen-induced arthritis(CIA).Methods Sixty BALB/c female mice were divided into the normal group,CIA model group,methotrexate(MTX)-positive control group,and Abatacept low-,medium-,and highdose groups according to the simple random grouping method,with 10 mice in each group.In addition to the normal group,the RA model was established by collagen induction in the remaining 5 groups.After CIA modeling,the mice were administered the equivalent dose of CIA according to the routine clinical dose for adult patients.The MTX positive control group was administered MTX tablets by gavage once per week.Abatacept low-,medium-and high-dose groups were intraperitoneally injected with the corresponding dose of Abatacept injection,200μL,once a week,respectively.The normal group and CIA model group were injected intraperitoneally with 0.9%sodium chloride solution,200μL,once a week.The arthritis index scores of each group were compared,and the expressions of serum TNF-α,IL-6 and matrix metalloproteinase(MMP)were detected by enzyme-linked immunosorbent assay,and the morphology of the synovial membrane of the ankle joint and the degree of inflammatory cell infiltration were observed by HE staining method;The expressions of TNF-α,osteopontin(OPN),NF-κB,and receptor activator of NF-κB ligand(RANKL)of the ankle joint were detected by immunohistochemistry method.Results Compared with the normal group,after modeling,the ankle joints of mice in all other groups were swollen,with severely limited mobility and elevated arthritis index scores;serum IL-6,TNF-α,MMP-2,and MMP-9 levels were elevated(all P<0.01);synovial hyperplasia and obvious inflammatory infiltration were observed in the ankle joints,and the expressions of TNF-α,OPN,NF-κB,and RANKL were all elevated.Compared with the CIA model group,the ankle joint swelling was reduced in the MTX positive control group and the Abatacept medium-and high-dose groups,the arthritis index scores were all also reduced(P<0.05),so were the expression levels of serum IL-6,TNF-α,MMP-2,and MMP-9(all P<0.01);synovial hyperplasia and inflammatory infiltration of the ankle joints were all significantly reduced,and the expression levels of TNF-α,OPN,NF-κB,and RANKL were all decreased.Conclusion Abatacept significantly attenuated joint inflammatory responses of CIA mice and inhibited their synovial proliferation and bone destruction.