α黑素细胞刺激素对CIA大鼠CD28和CD86分子及Th1/Th2型细胞因子的影响  

The effect of alpha-melanocyte-stimulating hormone on CD28 and CD86 molecule and Th1/Th2 cytokines in collagen-induced arthritis rats

作　　者：韩星海[1] 许倬[2] 田野苹[3] 闾坚强[1] 管剑龙[1] 蔡青[1] 施冶青[1] 

机构地区：[1]第二军医大学长海医院风湿免疫科,上海200433 [2]中国人民解放军二炮总医院肾内科 [3]第二军医大学基础部免疫教研室

出　　处：《中华风湿病学杂志》2004年第11期658-661,共4页Chinese Journal of Rheumatology

摘　　要：目的观察琢黑素细胞刺激素(琢-MSH)对胶原性关节炎(CIA)大鼠免疫细胞表达CD28和CD86分子及其产生Th1/Th2型细胞因子的影响,探讨α-MSH抑制CIA发病的相关免疫学机制。方法建立大鼠CIA动物模型,分离培养致敏大鼠脾细胞或腹股沟淋巴结细胞,加入琢-MSH(10-12～10-6mol/L)和(或)100μg/ml牛Ⅱ型胶原(bCⅡ),3H脱氧胸苷(TdR)掺入法测定琢-MSH对淋巴细胞增生反应的影响;收集上清,用酶联免疫吸附试验(ELISA)测定白细胞介素(IL)-1β、IL-10及干扰素(IFN)-γ含量,以L929为靶细胞,采用细胞毒活性四甲基偶氮唑蓝(MTT)显色法测定肿瘤坏死因子(TNF)-琢浓度,硝酸还原酶法测定一氧化氮(NO)分泌水平;应用荧光激活细胞分类(FACS)检测琢-MSH对T淋巴细胞表达CD28分子和巨噬细胞表达CD86分子的影响。结果琢-MSH能显著抑制bCⅡ诱导的CIA大鼠致敏淋巴细胞的增生反应,并显著抑制CIA大鼠致敏淋巴细胞表达CD28分子和腹腔巨噬细胞表达CD86分子;显著减少IFN-γ、IL-1和TNF-琢的产生,增加IL-10的分泌。结论琢-MSH有抑制CIA大鼠发病及抑制软骨破坏的作用,其机制与抑制CD28和CD86分子表达、调节细胞因子及NO的水平有关。Objective To explore immunological mechanism in the protective effect of α-melanocyte-stimulating hormone (MSH) on collagen-induced arthritis (CIA) in rats. Methods Immunized cells from in-guinal lymph nodes or spleen were cultured in the presence ofα-MSH (10-12～10-6 mol/L) and (or) 100 μg/ml bCⅡ. 3H thymidine was added and radioactivity incorporated into the cells. The effect ofα-MSH on the proliferation of lymphocytes was determined by liquid scintillation counting. Splenocytes and peritoneal macrophages from immunized rats were cultured withα-MSH (10-12～10-6 mol/L) and(or) 100 μg/ml bCⅡ(or 10 μg/ml LPS) . Culture supernatants were harvested and analyzed for interleukin (IL)-1β?interferon (IFN)-γ?IL-10 by ELISA. TNF-αwas detected by the L929 cytotoxicity using MTT colometric assay. NO was measured by the method of nitrate reductase. Flow cytometric analysis was performed to study the effects of α-MSH on the expressions of CD28 on the T-cells and CD86 on the macrophages. Results α-MSH inhibited the bCⅡ antigen-immunized T lymphocytes proliferation. Different concentrations ofα-MSH could suppress the production of IL-1β, IFN-γ, TNF-α, NO and enhance the synthesis of IL-10. The expression of CD28 on T lymphocyte and CD86 on macrophage were reduced byα-MSH administration. Conclusion α-MSH has a protective effect on CIA rats, with mechanisms of regulating the balance of Th1/Th2 cytokines, the production of NO and the expression of co-stimulatory molecules and inhibiting bCⅡ antigen-immunized T lymphocytes proliferation.

关 键 词：大鼠 CD86 CD28 Α-MSH 致敏 TH1/TH2型细胞因子 黑素细胞 分子 目的观 结论 

分 类 号：R593.22[医药卫生—内科学]