摘要目的 就GTP酶-Irgm1对巨噬细胞亚型M1分化的影响及可能的机制进行探讨.方法 分别对小鼠腹腔原代巨噬细胞和小鼠巨噬细胞系RAW264.7细胞进行培养,通过流式细胞术并以CD11b及F4/80标记巨噬细胞.每种细胞分成siRNA干扰组与对照组,然后待细胞贴壁后24 h加入干扰素(IFN)-γ和脂多糖(LPS)体外诱导巨噬细胞向M1转化,再进行流式细胞术iNOS染色鉴定M1的百分比.最后利用Western blot和RT-PCR检测体外诱导M1细胞过程时IRF5、IRF8及iNOS的表达情况.结果 ①加入Irgm1 siRNA干扰组的M1百分比(3.4％)明显低于对照组(35.8％),差异有统计学意义(P＜0.05);②QPCR检测结果显示:加入Irgm1 siRNA的干扰组与对照组相比,IRF5和IRF8 mRNA表达明显降低,差异有统计学意义(P＜0.05).Western blotting检测结果显示:加入Irgm1 siRNA干扰组与对照组相比,IRF5和IRF8蛋白表达明显减少,差异有统计学意义(P＜0.05).结论 巨噬细胞内Irgrn1能够通过影响IFR5和IRF8的表达影响巨噬细胞型别的改变.

abstractsPurpose To investigat the effect and possible mechanism of the GTPase-Irgm1 on macrophage M1differentiation.Methods Mouse peritoneal macrophages and mouse macrophage cell line RAW264.7 were cultured and employed as objects.CD11b and F4/80 were used to label mMacrophage (Mφ),which were testified by flow cytometry.Each kind of cells was used into two groups,siRNA interference group and control group.After 24 hours of adherence,IFN-gamma and LPS were added into the culture system to induce Mφ differentiation to M1.The percentage of M1 was identified by flow cytometry using iNOS staining.Finally,the expression of iNOS,IRF8 and IRF5 of induced M1 was detected by westernblot and RT-PCR.Results ①The percentage of M1 in the Irgm1 siRNA interference group was significantly lower than that in the control group.The difference was statistically significant (P ＜ 0.05);②Results obtained by RT-PCR test:Compared with the control group,the mRNA expression of IRF5 and IRF8 was significantly lower in the interference group.The difference was statistically significant (P ＜ 0.05).Results obtained by blotting Western test:Compared with the control group,the protein expression of IRF5 and IRF8 was significantly decreased in the Irgm1siRNA interference group.The difference was statistically significant (P ＜ 0.05).Conclusion Irgm1 in macrophage could have effect on the differentiation of subtype of macrophage (M1) through the expression of IRF5 and IRF8.