研究发现结核分枝杆菌(MTB)长期感染与巨噬细胞M2极化关系密切，其具体机制不明。小分子热休克蛋白Hsp16.3是MTB功能蛋白，对其在巨噬细胞中长期生存具有重要作用。我们前期研究发现MTB Hsp16.3可影响巨噬细胞功能。本项目拟建立体外诱导小鼠来源的巨噬细胞M1/M2极化技术平台，观察Hsp16.3对巨噬细胞M1/M2型极化的作用；构建敲减Hsp16.3的结核分枝杆菌株H37Rv感染的小鼠肺结核模型，动态观察体内肺泡巨噬细胞极化的改变和病理损伤，分析Hsp16.3表达水平与巨噬细胞极化的关系；采用cDNA芯片、RNA干扰和WB等技术进一步探讨TLR4等通路在Hsp16.3影响巨噬细胞M2 极化中的可能作用。项目预期结果可为后续深入探讨MTB 感染对巨噬细胞极化的影响及机制提供重要的实验和理论依据。

Recent studies showed that long-term infection of Mycobacterium tuberculosis (MTB) is closely associated with the polarization of M2 macrophages. However, its underlying mechanism remains unclear. Small heat shock protein Hsp16.3 is one of functional proteins of MTB, and plays an crucial role in the long-term survival of MTB in macrophages. Our previous study revealed that MTB Hsp16.3 played an important role in modulating the function of macrophage. This study aims to construct a technological platform for the induction of M1/M2 polarization of macrophages derived from mouse in vitro, and observe that the effects of MTB Hsp16.3 on M1/M2 polarization. Then, we further construct the Hsp16.3 knockdown strains of MTB H37Rv and establish the murine model of tuberculosis infection of lung. The changes of polarization of alveolar macrophage in vivo and the pathological injuries would be observed. The relationship between Hsp16.3 expression level and macrophage polarization would also be analyzed. Finally, the cDNA chip, RNA interference and Western BLot technology are used to explore the possible influence of the TLRs pathway including TLR4 on the effect of HSP16.3 on M2 polarization of macrophages. The expected results of this subject are to provide important experimental and theoretical basis for subsequent in-depth study on effects and mechanisms of the macrophage polarization by MTB infection.