仙茅苷下调ROS/NLRP3途径对大鼠骨髓单核细胞增殖及向破骨细胞分化的影响

Effect of curculigoside on proliferation and differentiation of bone marrow monocytes to osteoblasts through downregulation of the ROS/NLRP3 pathway

目的研究仙茅苷(curculigoside,CCG)对骨髓单核细胞体外增殖及分化的抑制作用。方法体外无菌分离大鼠骨髓单核细胞,通过细胞染色、周期分析评估CCG对巨噬细胞集落刺激因子(macrophage colony-stimulating factor,M-CSF)刺激的细胞增殖及核因子κB受体活化因子配体(receptor-activating factor ligand,RANKL)诱导分化的抑制效果。检测并分析活性氧(reactive oxygen species,ROS)/核苷酸结合寡聚化结构域样受体蛋白3(nucleotide-binding oligomerization domain-like receptor protein 3,NLRP3)途径相关成分的变化明确其作用途径和效果。结果CCG对骨髓单核细胞体外增殖有显著抑制作用,对细胞周期的作用在S期,并能诱导细胞凋亡(P<0.01)。当CCG干预浓度达到10-6 mol/L时,对骨髓单核细胞的体外破骨诱导有显著抑制作用。NLRP3、Cleaved casp-1、ROS、白介素18(interleukin-18,IL-18)、白介素1β(interleukin-1β,IL-1β)水平随着CCG干预浓度增大而降低(P<0.05)。构建NLRP3过表达质粒,转染骨髓单核细胞可逆转CCG在破骨诱导过程中对Cleaved casp-1的表达和IL-1β和IL-18水平的作用,使其上调(P<0.01)。结论CCG通过下调ROS/NLRP3途径抑制骨髓单核细胞增殖及向破骨细胞分化。

Objective To investigates how curculigoside(CCG)can potentially inhibit the proliferation and differentiation of bone marrow mononuclear cells in vitro.Methods Rat bone marrow mononuclear cells were aseptically isolated in a controlled in vitro environment.The impact of CCG on cell proliferation triggered by macrophage colony-stimulating factor(M-CSF)and on differentiation induced by receptor-activating factor ligand(RANKL)was evaluated through cell staining and cycle analysis.The changes in components linked to the reactive oxygen species/nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3(ROS/NLRP3)pathway were detected and analyzed.These components encompassed ROS,NLRP3,Cleaved caspase-1(Cleaved casp-1),interleukin-1β(IL-1β),and interleukin-18(IL-18).Results In vitro,CCG exhibited a substantial inhibitory effect on bone marrow mononuclear cell proliferation,prompting cell cycle arrest in the's phase and inducing apoptosis(P<0.01).The treated cells reduced expression levels of NLRP3,Cleaved casp-1,ROS,IL-18,and IL-1β,with the reduction becoming more pronounced as CCG concentrations increased(P<0.05).The introduction of a NLRP3 overexpression plasmid and subsequent transfection into bone marrow mononuclear cells counteracted CCG's impact on induced differentiation.This resulted in the upregulation of Cleaved casp-1 expression,as well as elevated levels of IL-1βand IL-18(P<0.01).Conclusion CCG suppressed the proliferation and osteoblast differentiation of bone marrow monocytes by reducing activity in the ROS/NLRP3 pathway.