Chen Cong, Huang Hai
Department of Clinical Trial Ward Outpatient Office, Wuhan Pulmonary Hospital, Wuhan, Hubei, China.
Department of TB Ward II, Wuhan Pulmonary Hospital, Wuhan, Hubei, China.
Cytojournal. 2025 Apr 1;22:38. doi: 10.25259/Cytojournal_144_2024. eCollection 2025.
(MTB) manipulates macrophage functions, thus mediating tuberculosis (TB) progression. Whether the switch/sucrose non-fermenting-related matrix-associated actin-dependent regulator of chromatin subfamily a member 5 (SMARCA5) mediates MTB-induced macrophage polarization remains unclear.
Human Promyelocytic Leukemia Cell Line was induced into macrophages and then treated with MTB. Cell viability and apoptosis were tested with cell counting kit 8 assay and flow cytometry. Classically activated macrophages (M1) polarization and inflammation were measured by detecting CD86 cell rate and inflammatory factor levels. The levels of SMARCA5, methyltransferase 3 (METTL3), and insulin-like growth factor 2 binding protein 1 (IGF2BP1) were assessed using quantitative real-time polymerase chain reaction or Western blot. The interaction between SMARCA5 and METTL3 or IGF2BP1 was confirmed by methylated RNA immunoprecipitation (RIP) and RIP assays. The effect of METTL3 knockdown on SMARCA5 messenger RNA (mRNA) stability was evaluated using actinomycin D treatment.
MTB treatment suppressed the viability and promoted the apoptosis and M1 polarization and inflammation of macrophages ( < 0.05), and SMARCA5 knockdown abolished these effects ( < 0.05). METTL3 mediated the m6A methylation of SMARCA5 to enhance the mRNA stability of the latter, and this modification was recognized by IGF2BP1. SMARCA5 upregulation reverted the si-METTL3-mediated inhibition of MTB-induced macrophage M1 polarization and inflammation ( < 0.05).
METTL3-mediated SMARCA5 facilitates macrophage M1 polarization and inflammation, providing a novel target for TB treatment.
结核分枝杆菌（MTB）可操纵巨噬细胞功能，从而介导结核病（TB）进展。开关/蔗糖非发酵相关基质相关肌动蛋白依赖性染色质调节因子a亚家族成员5（SMARCA5）是否介导MTB诱导的巨噬细胞极化尚不清楚。
将人早幼粒细胞白血病细胞系诱导为巨噬细胞，然后用MTB处理。用细胞计数试剂盒8检测法和流式细胞术检测细胞活力和凋亡情况。通过检测CD86细胞率和炎症因子水平来测定经典活化巨噬细胞（M1）极化和炎症。使用定量实时聚合酶链反应或蛋白质免疫印迹法评估SMARCA5、甲基转移酶3（METTL3）和胰岛素样生长因子2结合蛋白1（IGF2BP1）的水平。通过甲基化RNA免疫沉淀（RIP）和RIP检测证实SMARCA5与METTL3或IGF2BP1之间的相互作用。使用放线菌素D处理评估METTL3敲低对SMARCA5信使核糖核酸（mRNA）稳定性的影响。
MTB处理抑制了巨噬细胞的活力，促进了其凋亡、M1极化和炎症（<0.05），而SMARCA5敲低消除了这些影响（<0.05）。METTL3介导SMARCA5的m6A甲基化以增强后者的mRNA稳定性，并且这种修饰被IGF2BP1识别。SMARCA5上调逆转了si-METTL3介导的对MTB诱导的巨噬细胞M1极化和炎症的抑制作用（<0.05）。
METTL3介导的SMARCA5促进巨噬细胞M1极化和炎症，为结核病治疗提供了一个新靶点。
