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目的研究铜绿假单胞菌分泌的N-3-氧代十二烷酰-L-同型丝氨酸内酯(N-3-oxododecanoyl-L-homoserine lactone,3-O-C12-HSL)是否通过过氧化物酶增殖物激活受体γ(peroxisome proliferater-activated receptorγ,PPARγ)调节单核细胞来源的树突细胞(monocytes derived-dendritic cells,Mo-DCs)表达白细胞介素6(interleukin-6,IL-6)。方法 PPARγ配体筛选实验体外检测3-O-C12-HSL是否与PPARγ结合。免疫磁珠法分选人CD14+单核细胞,人白细胞介素4(human Interleukin 4,h IL-4)及人粒单核细胞集落刺激因子(human granulocyte monocyte colony stimulating factor,h GM-CSF)诱导分化为Mo-DCs,不同浓度的3-O-C12-HSL处理Mo-DCs;PPARγ拮抗剂GW9662处理Mo-DCs后,3-O-C12-HSL处理Mo-DCs,电化学发光法检测Mo-DCs培养上清IL-6浓度。结果配体筛选实验显示3-O-C12-HSL>4.60μmol/L时可竞争性结合PPARγ配体结合区。3-O-C12-HSL下调脂多糖(lipopolysaccharide,LPS)诱导的Mo-DCs表达IL-6水平(P<0.05),PPARγ特异性拮抗剂GW9662能部分逆转3-O-C12-HSL介导的IL-6表达下调(P<0.05)。结论 3-O-C12-HSL可与PPARγ相结合,并通过PPARγ调节Mo-DCs分泌表达IL-6。
Objective To investigate whether N-3-oxododecanoyl-L-homoserine lactone (3-O-C12-HSL) secreted by Pseudomonas aeruginosa Peroxisome proliferator-activated receptorγ (PPARγ) regulates the expression of interleukin-6 (IL-6) in monocytes derived-dendritic cells (Mo-DCs) ). Methods PPARγ ligand screening assay was used to detect whether 3-O-C12-HSL binds to PPARγ in vitro. Immunomagnetic beads were used to select human CD14 + monocytes, human interleukin 4 (hIL-4) and human granulocyte monocyte colony stimulating factor (h GM-CSF) Mo-DCs were treated with different concentrations of 3-O-C12-HSL, Mo-DCs were treated with 3-O-C12-HSL treated with PPARγ antagonist GW9662, and the chemiluminescence method was used to detect Mo- DCs culture supernatant IL-6 concentration. Results Ligand screening experiments showed that 3-O-C12-HSL> 4.60 μmol / L competitively bound PPARγ ligand binding domain. 3-O-C12-HSL downregulated the expression of IL-6 in Mo-DCs induced by lipopolysaccharide (LPS) (P <0.05), and PPARγ-specific antagonist GW9662 could partially reverse 3-O-C12-HSL mediated IL-6 expression was down-regulated (P <0.05). Conclusion 3-O-C12-HSL can bind to PPARγ and regulate the secretion of IL-6 by Mo-DCs via PPARγ.