苜草素对巨噬细胞吞噬活性和NO、IL-6、TNF-α分泌的影响

EFFECTS OF POLYSAVONE ON PHAGOCYTIC ACTIVITY AND NO,IL-6,AND TNF-α PRODUCTION OF MACROPHAGES

目的研究苜草素对小鼠巨噬细胞RAW264.7吞噬活性和NO、IL-6、TNF-α的影响,探讨苜草素对巨噬细胞免疫功能的影响机制。方法以不同浓度的苜草素和10 g/ml的脂多糖(LPS)分别处理巨噬细胞RAW264.7,采用中性红法测定巨噬细胞的吞噬活性,ELISA检测TNF-α分泌量。以不同浓度的苜草素处理LPS诱导的巨噬细胞RAW264.7,采用Griess分析法测定NO含量,ELISA检测IL-6分泌量,半定量RT-PCR方法测定巨噬细胞中iNOS、IL-6、TNF-αmRNA水平。结果苜草素在0.4、0.6、0.8和1.0 mg/ml时能显著提高巨噬细胞RAW264.7的吞噬活性(P<0.05);在0.6、0.8和1.0 mg/ml时显著增加TNF-α的分泌量(P<0.01),显著降低LPS诱导的巨噬细胞NO产生量(P<0.05);在0.2、0.4、0.6、0.8和1.0 mg/ml时显著降低LPS诱导的巨噬细胞IL-6的产生量(P<0.01)。苜草素可抑制LPS诱导的iNOS mRNA和IL-6 mRNA的表达,增加TNF-αmRNA的表达。结论苜草素可以提高巨噬细胞RAW264.7的吞噬活性,在转录水平上影响小鼠巨噬细胞iNOS、IL-6、TNF-α的基因表达,对巨噬细胞免疫功能起调节作用。

Objective To investigate the effects of polysavone on phagocytic activity and NO,IL-6,and TNF-α production of macrophage RAW264.7 in vitro.Method Macrophages were treated with different concentration of polysavone and LPS（10 μg/ml） respectively.The phagocytic activity of macrophage was detected by neutral red.TNF-α in culture supernatants was measured by ELISA.NO and IL-6 production were detected by Griess and ELISA respectively.The iNOS,IL-6,and TNF-α mRNA levels of macrophage were measured with RT-PCR.Results The phagocytic activity of macrophage in the polysavone group（0.4,0.6,0.8 or 1.0 mg/ml） was significantly greater than that of the control（P0.05）.The production of TNF-α was increased significantly,but LPS-induced NO production was decreased significantly by polysavone（0.6,0.8 and 1.0 mg/ml,P0.01,P0.05）.Polysavone caused significant inhibition on LPS-induced IL-6 production（0.2,0.4,0.6,0.8 or 1.0 mg/ml,P0.01）.Polysavone inhibited LPS-induced production of iNOS and IL-6 mRNAs,and increased production of TNF-α mRNA.Conclusion Polysavone appears to exert immune modulating effect by regulating the phagocytic activity of macrophage RAW 264.7,the expression of IL-6,iNOS and TNF-α,as well as the production of NO.