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目的 研究α 促黑素细胞激素 (α MSH)对巨噬细胞产生炎症相关的免疫分子的影响 ,探讨α MSH的抗炎作用机制。方法 分别用LPS或α MSH +LPS处理体外培养的小鼠巨噬细胞系RAW2 6 4.7细胞 ,以Griess试剂测定NO ,采用DADPH 黄递酶组织化学染色法检测iNOS ,采用MTT法检测IL 1和TNF α ,采用ELISA法测定IL 6 ,细胞膜表面免疫相关分子的表达用FACS进行分析。结果 RAW2 6 4.7细胞在LPS刺激下产生NO、IL 1、IL 6和TNF α显著增多 ,iNOS活性明显增强 ,细胞表面MHCⅡ、CD14、CD40、CD5 4、CD80和CD86分子表达水平增高 ;若在LPS刺激的同时给予α MSH ,能够抑制LPS诱导RAW2 6 4.7细胞产生NO ,IL 1、IL 6和TNF α ,使iNOS活性及表达MHCⅡ、CD14、CD40、CD5 4、CD86分子降低 ,但CD80分子降低不明显。结论 α MSH抑制巨噬细胞产生NO、iNOS、前炎性细胞因子及表达膜表面免疫相关分子是其抗炎作用的重要机制之一
Objective To investigate the effect of α-melanocyte stimulating hormone (α-MSH) on immune-related immune molecules produced by macrophages and to explore the anti-inflammatory mechanism of α-MSH. Methods RAW264 cells cultured in vitro were treated with LPS or α MSH + LPS, respectively. NO was detected by Griess reagent, and iNOS was detected by DADPH diaphorase histochemical staining. IL - 1 and TNF α, IL 6 was detected by ELISA, and the expression of immune-related molecules on cell membrane surface was analyzed by FACS. Results NO2, IL-1, IL-6 and TNFα were significantly increased in LPS-stimulated RAW264.7 cells and the iNOS activity was significantly increased. The expression of MHCⅡ, CD14, CD40, CD54, CD80 and CD86 on the cell surface was increased. Stimulation of α MSH at the same time can inhibit LPS-induced NO, IL 1, IL 6 and TNFα production in RAW264.7 cells, decrease iNOS activity and expression of MHCⅡ, CD14, CD40, CD54, and CD86 molecules, but decrease CD80 molecules obvious. Conclusion α-MSH can inhibit the production of NO, iNOS, pro-inflammatory cytokines and immune-related molecules on membrane surface of macrophages, which is one of the important mechanisms of anti-inflammatory effects