茶树菇多糖对小鼠巨噬细胞RAW264.7免疫功能的影响

Effects of Agrocyb aegerita Polysaccharid on Immune Function of Mouse Macrophages RAW264.7

为了揭示茶树菇多糖(AAP)在体外对小鼠巨噬细胞RAW264.7免疫功能的影响,本试验以RAW264.7为研究对象,将其分为空白对照(CON)组、AAP低剂量(AAPL)组、AAP中剂量(AAPM)组、AAP高剂量(AAPH)组和脂多糖(LPS)组,采用噻唑蓝(MTT)比色法检测AAP对巨噬细胞RAW264.7的安全浓度;流式细胞仪检测巨噬细胞RAW264.7表面协同刺激分子抗原分化簇80(CD80)和86(CD86)的表达,酶联免疫吸附试验(ELISA)检测细胞因子白介素-1β(IL-1β)、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)和一氧化氮(NO)的浓度。结果显示,AAP浓度低于250μg/mL,不会对小鼠巨噬细胞RAW264.7产生毒性作用;与CON组相比,AAPM组和AAPH组RAW264.7细胞的表面分子CD80及所有AAP组RAW264.7细胞的表面分子CD86均极显著升高(P<0.01或P<0.001);AAPH组IL-1β及AAPM组和AAPH组IL-6、TNF-α和NO表达水平均显著增加(P<0.05或P<0.01或P<0.001)。结果表明,AAP能提高小鼠巨噬细胞RAW264.7表面分子CD80和CD86表达水平及细胞因子IL-1β、IL-6、TNF-α和NO的分泌能力。

In order to elucidate the effects of Agaricus auricula polysaccharides(AAP)on the immune function of mouse macrophages RAW264.7 in vitro,this study utilized RAW264.7 cells and divided them into blank control(CON)group,low-dose AAP(AAPL)group,medium-dose AAP(AAPM)group,high-dose AAP(AAPH)group,and lipopolysaccharide(LPS)group.The MTT colorimetric assay was employed to determine the safe concentration of AAP for RAW264.7 cells.Flow cytometry was used to detect the expression of surface co-stimulatory molecules CD80 and CD86 on RAW264.7 cells,while enzyme-linked immunosorbent assay(ELISA)was employed to measure the concentrations of cytokines interleukin-1β(IL-1β),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and nitric oxide(NO).The results demonstrated that AAP concentrations below 250μg/mL were non-toxic to mouse macrophages RAW264.7.Compared with the CON group,the expression levels of surface molecule CD80 on RAW264.7 cells significantly increased in the AAPM and AAPH groups,as well as CD86 in all AAP groups(P<0.01 or P<0.001).The expression levels of IL-1βin the AAPH group and IL-6,TNF-α,and NO in the AAPM and AAPH groups exhibited significant elevation(P<0.05,P<0.01,or P<0.001).The results indicate that AAP enhances the expression levels of surface molecules CD80 and CD86,and promotes the secretion of cytokines IL-1β,IL-6,TNF-α,and NO in mouse macrophages RAW264.7.