| Objective:To investigate the effect of low molecular weight of Agaricus blazei Murill ploysaccharide(ABP-AWA1)isolated and purified from Agaricus blazei Murill on phagocytosis and secretional function of macrophages(Mφ)in mouse peritoneal,so as to detect the regulation effect of ABP-AWA1 on immune functionMethods:Kunming mice was injected by intraperitoneal with 1 mL 5%soluble starch,Mφ were isolated and purified from mice peritoneal after 3 days.The cells were cultured in RPMI-1640 as blank control group,treated with lipopolysaccharide(Lipopolysaccharide,LPS)as the positive control group,and treated with different concentration of A BP-AWA1(75 μg/mL、150μg/mL、300μg/mL)as drug treatmental groups.Phagocytic function of Mφ in mice peritoneal was determined by neutral red method;NO secretion was determined by Griess reaction method;Enzyme-linked immunosorbent assay(ELISA)was used to measure the secretions of NO,IL-1β,IL-6,IL-1 2p70 and TNF-α in different times under 150 μg/mL ABP-AWA1;mRNA expressions of IL-1β、IL-6、IL-12p40 and TNF-α were detected by real-time quantitative PCR.Results:The test of Mφ phagocytic ability showed that ABP-AWA1 significantly improved the neutral red phagocytic ability of Mφ in mice peritoneal(P<0.05),and this effect was most obvious at the concentration of 150 μg/mL.Griess reaction test found that ABP-AWA1 increased the release of NO(P<0.05),and this effect was most obvious at the concentration of 150 μg/mL,and this release of NO was stable at 48 h.The result of ELISA showed the cytokines IL-1β,IL-6,IL-12p70 and TNF-α were increased obviously after the treatments of ABP-AWA1(P<0.05),simultaneously in the study of culture time on cytokine secretion,we found the increasements of IL-1β,IL-6 and IL-12p70 were undetectable after 48h-treatment(P>0.05),and the peak time of TNF-α expression was appeared at 24 h.RT-PCR showed that ABP-AWA1 significantly enhanced the mRNA expressions of cytokines IL-1β,IL-6,IL-12p40 and TNF-α.Conclusion:1.ABP-AWA1 could stimulate the activation of macrophages in mouse peritoneal,and enhanced the phagocytic capacity,thereby enhancing the body’s immunity.2.ABP-AWA1 could increase the secretion of NO and cytokines IL-1,IL-6,IL-12p70,TNF-α in mouse peritoneal macrophages,and the levels of IL-1,IL-6,IL-12p40,TNF-α mRNA were increased after the treatments of ABP-AWA1,suggesting the regulation of secretion and expression of cytokines refered to the immune regulation. |
