MTB Hsp 16.3通过TLR4对小鼠M1型巨噬细胞的作用研究  

MTB Hsp16.3 affects the mouse M1 macrophage through TLR4

作　　者：高绍莹 秦欢[1] 李姗姗[1] 刘芊伊 泮红飞 刘旭恒[1] 龙润莹 张焱皓 徐林[1] 罗军敏[1] 

机构地区：[1]遵义医学院免疫学教研室贵州省免疫分子应用研究工程中心,遵义563099

出　　处：《现代免疫学》2017年第5期366-373,共8页Current Immunology

基　　金：国家自然科学基金地区项目(81460249)

摘　　要：探讨MTB Hsp16.3通过TLR4对小鼠M1型巨噬细胞的作用。从BALB/c小鼠胫腓骨取骨髓来源巨噬细胞,经IFN-γ诱导得到M1型巨噬细胞,MTB Hsp16.3与M1型巨噬细胞共培养,qRT-PCR和ELISA检测M1/M2型巨噬细胞相关细胞因子表达水平;用siRNA-TLR4和PMB抑制M1型巨噬细胞,qRT-PCR和ELISA检测IL-10、Arg1、iNOS、TGF-β及TNF-α等细胞因子的表达水平,western blotting法检测MAPK-NF-κB信号通路中各蛋白的表达水平。结果发现MTB Hsp16.3作用后的M1型巨噬细胞和M2型巨噬细胞的细胞因子IL-10、TGF-β及Arg1mRNA在各时间点均升高,M1型细胞因子TNF-α、iNOS表达水平降低。抑制TLR4后,IL-10、Arg1及TGF-β水平降低,TNF-α、iNOS、IL-6表达水平升高,MAPK-NF-κB信号途径被抑制。由此MTB Hsp16.3可能通过TLR4促进小鼠M1型巨噬细胞发生M2样转化。We aimed to investigate the signal pathway by which Hsp16.3affects phenotype of mouse macrophages.Bone marrow cells were extracted from the tibia and fibular of BALB/c mice.IFN-γwas used to induce M1 macrophage from macrophage M0.M1 macrophages were cultured with MTB Hsp16.3.Subsequently,the expressions of M1/M2 cytokines were detected by quantitative RT-PCR and ELISA.After the TLR4 in M1macrophages were inhibited by siRNA-TLR4 and PMB,the M1 macrophages were stimulated with MTB Hsp16.3,and the expressions of Arg1,IL-10,iNOS,TGF-βand TNF-αwere determined by quantitative RT-PCR and ELISA.The expression of the MAPK-NF-κB signaling pathway was detected by western blotting.The results showed that in MT13Hsp16.3stimulated M1 mocrophages,the expression of M2 cytokines IL-10、TGF-βand Arg1 mRNA were increased,and the expressions of M1 cytokines iNOS and TNF-αwere decreased.After the TLR4 was inhibited in M1 macrophages,the effects of MTB Hsp16.3were declined and the expression of the MAPK-NF-κB signaling pathway was concomitantly declined.The results indicate that MTB Hsp16.3makes the conversion of M2 macrophages from the M1-like macrophages through the MAPK-NF-κB signaling pathway mediated by TLR4.

关 键 词：MTB HSP16.3 M1型巨噬细胞 TLR4 

分 类 号：R392.11[医药卫生—免疫学]