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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 investigate the effects of melanocyte stimulating hormone (MSO) on the expression of CD28 and CD86 and Th1 / Th2 type cytokines in immune cells of collagen-induced arthritis (CIA) rats and to explore the role of α-MSH in inhibiting the pathogenesis of CIA Related immunological mechanisms. Methods The rat CIA model was established and the sensitized rat splenocytes and inguinal lymph node cells were isolated and cultured. The cells were cut into MSH (10-12 ~ 10-6mol / L) and / or 100μg / ml bCⅡ , 3H-thymidine (TdR) incorporation method was used to determine the effect of MSH on lymphocyte proliferation. The supernatants were harvested and the levels of IL-1β, IL-10 and IL-10 were measured by enzyme linked immunosorbent assay Interferon (IFN) -γ content, L929 as target cells, cytotoxicity of MTT was used to determine the concentration of tumor necrosis factor (TNF) - cut nitrate, nitric acid reductase method for the determination of monoxide (NO) secretion. The effect of MSH on the expression of CD28 and CD86 on T lymphocytes was detected by fluorescence activated cell sorting (FACS). RESULTS: MSH could significantly inhibit the proliferation of sensitized lymphocytes induced by bCII in CIA rats, and significantly inhibit the expression of CD28 molecules in sensitized lymphocytes of CIA rats and the expression of CD86 molecules in peritoneal macrophages. The levels of IFN-γ, IL -1 and TNF-α production, increase IL-10 secretion. Conclusion: MSH can inhibit the onset of CIA rats and inhibit the destruction of cartilage, the mechanism of which is related to inhibiting the expression of CD28 and CD86, regulating the levels of cytokines and NO.