Objective To study the effect of supernatant of B16F10 melanoma cell culture on the release of tumor necrosis factor-α (TNF-α) and nitrogen monoxide (NO) from mouse peritoneal macrophages stimulated by lipopolysaccharide (LPS). Methods Syngeneic mouse peritoneal macrophages were cultured in vitro for 24 hours with the induction by LPS in two groups : the experimental group in which the supernatant of B16F10 cell culture was used, and blank control group in which a culture medium, RPMI 1640, was used instead. The immunocytochemistry assay and cytotoxicity on L929 cells with methyl thiazolyl tetrazolium (MTT) assay were applied for the detection of TNF-α expression and activity, and the Griess reaction assay was used for NO level determination. Results In the assay, the positive expression of TNF-α was obviously lower in experimental group than that in blank control group, and the activity of TNF-α in experimental group was significantly lower than that in blank control group [ absorbance (A) value : 0.746±0.049 vs. 0.387±0.030.P＜0.01) ; the level of NO released by macrophages was also lower significantly in experimental group than that in blank control group (μmol/L : 4.830±1.384 vs. 11.455±0.175, P＜0.01). Conclusion The supernatant of B16F10 melanoma cell culture has the ability to suppress the secretion of TNF-α and NO in the syngeneic mouse peritoneal macrophages induced by LPS.%目的 探讨B16F10黑素瘤细胞培养上清液对脂多糖(LPS)刺激小鼠腹腔巨噬细胞释放肿瘤坏死因子-α (TNF-α)和一氧化氮(NO)的影响.方法 体外培养同基因小鼠腹腔巨噬细胞,分为实验组和空白对照组.实验组加入B16F10黑素瘤细胞培养上清液,空白对照组加入RPMI 1640完全培养基,分别用LPS刺激后继续培养24 h,采用免疫细胞化学染色法及L929细胞抑制实验[四甲基偶氮唑盐(MTT)比色法]检测巨噬细胞TNF-α的表达及活性,Griess法检测NO的含量.结果 实验组TNF-α阳性表达较空白对照组明显减少,TNF-α活性较空白对照组明显降低[吸光度(A值):0.746±0.049比0.387±0.030,P＜0.01]；巨噬细胞释放NO的水平明显低于空白对照组(μmol/L:4.830± 1.384比11.455±0.175,P＜0.01).结论 B16F10黑素瘤细胞培养上清液对LPS诱导的同基因小鼠腹腔巨噬细胞分泌TNF-α、NO具有抑制作用.
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