NLRP3炎性小体对THP-1细胞吞噬老化红细胞后亚类分化的调控

Modulation of NLRP3 Inflammasome Activity in Subtype Differentiation of THP-1 Cells after Phagocytosis of Aged Erythrocyte

目的明确NLRP3炎性小体是否可调节巨噬细胞对红细胞的吞噬能力,调控巨噬细胞的亚类分化。方法以NLRP3低表达的THP-1细胞(ID3 THP-1),转空载体的THP-1细胞(shNC THP-1)及野生型THP-1细胞模拟巨噬细胞,与未处理红细胞、水浴老化红细胞、IgG致敏红细胞分别孵育。流式细胞术检测THP-1细胞对不同红细胞的吞噬率;流式细胞术检测巨噬细胞M1亚类指标CD16和CD86,M2亚类指标CD163和CD206在THP-1细胞上的表达。结果NLRP3低表达的ID3 THP-1对红细胞的吞噬能力明显下调。老化红细胞具有诱导THP-1向M1亚类分化,抑制THP-1向M2亚类分化的能力。当THP-1的NLRP3炎性小体表达下降时,吞噬红细胞后,其向M1亚类分化的能力下调,而向M2亚类分化的能力增强。结论NLRP3炎性小体可作为一个靶向调节位点,调控巨噬细胞对红细胞吞噬,及其自身的亚类分化。

Objective To clarify whether NLRP3 inflammasome can modulate macrophage phagocytic capacity towards red blood cells(RBCs)and regulate macrophage subtype polarization.Methods THP-1 cells with low expression of NLRP3(ID3 THP-1),cells transfected with an empty vector(shNC THP-1),and wild-type THP-1 cells were used as macrophage models.These cells were incubated with untreated RBCs,water bath-aged RBCs,and IgG-opsonized RBCs,respectively.Flow cytometry was used to detect the phagocytic rate of THP-1 cells to different RBC types and to measure the expression of M1 subtype markers(CD16 and CD86),and M2 subtype markers(CD163 and CD206)on THP-1 cells.Results ID3 THP-1 with reduced NLRP3 expression exhibited significantly downregulated phagocytic capacity towards RBCs.Aged RBCs induced the differentiation of THP-1 cells into the M1 subclass while inhibiting their differentiation into the M2 subclass.Decreased expression of the NLRP3 inflammasome in THP-1 cells led to a downregulation of their ability to differentiate into the M1 subclass following RBC phagocytosis,accompanied by their enhanced capacity to differentiate into the M2 subclass.Conclusion NLRP3 inflammasome can serve as a pivotal regulatory target,governing macrophage phagocytosis of RBCs and their subsequent subclass differentiation.