TLR4抗体对姬松茸多糖作用巨噬细胞产生细胞因子的影响  

INFLUENCES OF TLR4 ANTIBODY ON AGARICUS BLAZEI MURILL POLYSACCHARIDES EFFECT IN RAW264.7 CELLS

作　　者：李晨光[1] 冯翠萍[1] 常明昌[1] 孟俊龙[1] 欧阳玉倩[1] 

机构地区：[1]山西农业大学食品科学与工程学院,太谷030801

出　　处：《营养学报》2015年第5期490-494,共5页Acta Nutrimenta Sinica

基　　金：山西省自然科学基金(No.2010011043-1);山西农业大学博士科研启动经费项目(No.412564);山西省科技攻关项目(No.021025);山西省科技攻关项目(No.20130311017-3)

摘　　要：目的研究TLR4抗体对巴氏蘑菇多糖(Agaricus blazei Murrill polysaccharide,ABPS)对体外巨噬细胞RAW264.7产生细胞因子白介素-1β(IL-1β)、肿瘤坏死因子--α(TNF-α)、干扰素-β(IFN-β)的影响。方法相同浓度ABPS及脂多糖(LPS,阳性对照)作用巨噬细胞RAW264.7 3 h、6 h、12 h、18 h和36 h;不同浓度ABPS及LPS作用巨噬细胞RAW264.7 24 h;TLR4抗体作用巨噬细胞RAW264.7 1 h后,更换含有ABPS和LPS的细胞培养液作用24h。收集细胞培养上清,采用ELISA法检测细胞培养上清细胞因子IL-1β、TNF-α、IFN-β的含量。结果 ABPS作用巨噬细胞RWA264.7 24 h后,细胞培养上清中IL-1β、TNF-α、IFN-β含量明显高于阴性对照,差异有统计学意义(P<0.01),但显著低于LPS组。在一定浓度范围内,细胞培养上清中IL-1β、TNF-α、IFN-β的含量随ABPS浓度增大而增大,浓度为1000μg/ml时IL-1β、TNF-α、IFN-β含量达到最大。TLR4抗体处理的巨噬细胞RAW264.7经ABPS和LPS作用,IL-1β、TNF-α、IFN-β含量低于未处理组,差异有统计学意义(P<0.01)。结论 TLR4抗体能够一定程度抑制ABPS作用巨噬细胞RAW264.7产生IL-1β、TNF-α、IFN-β,推测ABPS可能通过TLR4受体信号转导通路影响三种细胞因子的释放。Objective To study the effects of TLR4 antibody on Agaricus Blazei murill polysaccharides （ABPS） action role on cytokines IL-1β, TNF-α, IFN-β production in RAW264.7 cells. Methods Macrophage RAW264.7 was treated with same concentration of lipopolysaccharide （LPS） and ABPS for 3, 6, 12, and 36 h, or different concentrations of LPS and ABPS for 24 h, in the presence or absence of TLR4 antibody for 1 hour. Cell culture supernatants were collected, and the content of eytokines IL-1β, TNF-α, IFN-β were detected by ELISA. Results After treated by ABPS for 24 h, the IL-1β, TNF-α, IFN-β cytokines in the cell culture supernatants were significantly higher than negative control group （P〈0.01）, but significantly lower than positive control （LPS）. In the certain concentration range of ABPS, the IL-1β, TNF-α, IFN-β production in cell culture supernatants increased with the concentration of ABPS. The IL-1β, TNF-α, IFN-β production were up to the maximum when treated with 1000 ug/ml ABPS. After treated with TLR4 antibody, the IL-1β, TNF-α, IFN-β contents were significantly lower than untreated groups （P〈0.01）. Conelution TLR4 can inhibit the effect of ABPS on IL-1β, TNF-α, IFN-β cytokines production in RAW264.7 cells. We spectulate that the effects of ABPS on cytokines release may be through TLR4 receptor signal transduction pathway.

关 键 词：巴氏蘑菇多糖 巨噬细胞RAW264.7 TLR4抗体 细胞因子 

分 类 号：R285.5[医药卫生—中药学]