BAFF-BAFFR及信号转导分子参与佐剂性关节炎的免疫反应及芍药苷-6′-O-苯磺酸酯的作用  

Paeoniflorin-6'-O-tosylate ameliorate the immune response in adjuvant arthritis rat through BAFF-BAFFR downstream signal molecule

作　　者：徐澍[1] 魏芳[1] 贾晓益[1] 孙晓静[1] 杨雪枝 赵英杰[1] 常艳[1] 魏伟[1] 

机构地区：[1]安徽医科大学临床药理研究所、抗炎免疫药物教育部重点实验室、安徽抗炎免疫药物协同创新中心,合肥230032

出　　处：《安徽医科大学学报》2016年第5期643-649,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金资助项目(编号:81330081,81573443,31200675)

摘　　要：目的观察佐剂性关节炎(AA)大鼠炎症不同时期脾脏B淋巴细胞刺激因子(BAFF)及其受体(BAFFR)表达变化以及芍药苷-6′-O-苯磺酸酯(CP-25)对其的影响。方法采用完全弗氏佐剂制备大鼠AA模型,随机分为正常组、AA组、CP-25(25、50、100 mg/kg)组、甲氨蝶呤(0.5 mg/kg)组、白芍总苷(50 mg/kg)组、芍药苷(50 mg/kg)组,造模后第17~31天给药。ELISA法检测外周血BAFF水平;免疫组织化学法、流式细胞术、Q-PCR和Western blot法检测CP-25对脾脏BAFFR表达的影响;Western blot法检测脾脏肿瘤坏死因子相关因子2(TRAF2)蛋白的表达。结果与正常组比较,炎症反应期(D9)、炎症初期(D16)、炎症高峰期(D23)及炎症缓解期(D30)模型大鼠血清BAFF水平和脾脏BAFFR mRNA及蛋白表达均增加。与AA组比较,CP-25(25、50、100 mg/kg)体内给药明显下调AA大鼠脾脏BAFFR mRNA及蛋白表达。体外BAFF刺激后,大鼠脾脏细胞TRAF2及BAFFR蛋白表达显著增加,CP-25(1×10^(-6)mol/L)明显降低TRAF2及BAFFR蛋白的表达。结论 CP-25发挥炎症免疫调节作用可能与其抑制AA大鼠BAFFR受体及TRAF2信号分子表达有关。Objective To observe the expression profile of splenic B lymphocyte stimulating factor receptor（ BAF- FR） in adjuvant arthritis（AA） rats during the inflammation course of arthritis and the effects of Paeoniflorin -6＇-0- tosylate （ code ： CP-25 ）. Methods AA rats were prepared using complete freund＇s adjuvant and were divided into 8 groups randomly, including normal, AA, CP-25 （25, 50, 100 mg/kg）, methotrexate （0.5 mg/kg）, total gluco- sides of paeony （TGP） （50 mg/kg）, and paeoniflorin （Pae） group （ 50 mg/kg）, and treated these groups from dl 7 to d31 after preparation of AA model. BAFF levels in peripheral blood were detected by ELISA; the expression of BAFFR during the inflammation course of AA rat in spleen was detected by flow cytometry. The effect of CP-25 on BAFFR in spleen was investigated through immunohistochemistry, Q-PCR and Western blot. The expression of tumor necrosis factor receptor associated factor （TRAF2） in rat spleen were detected by Western blot. Results Compared with the normal, the levels of BAFF in serum were increased in d9, d16, d23 and d30 AA model and the expressions of BAFFR mRNA and protein also increased obviously. CP-25 （25,50,100 mg/kg） could decrease the expression of BAFFR mRNA. In vitro, with the stimulation of BAFF, the expressions of TRAF2 and BAFFR in splenocyte were increased apparently, and CP-25 （ 1 x 10 -6 mol/L） could decrease their expression significantly.Conclusion CP-25 play important roles in immune regulation in AA rats through reducing the expression of BAF- FR and TRAF2 in spleen.

关 键 词：类风湿关节炎 佐剂性关节炎 CP-25 BAFF BAF-FR TRAF2 

分 类 号：R967[医药卫生—药理学]