摘要目的 研究多聚肌苷-多聚胞苷酸钠盐[polyinosinic polycytidylic acid,Poly(I:C)]的不同处理方式对3T3-L1脂肪细胞固有免疫应答的影响.方法 将Poly(I:C)通过外源直接加入和转染两种方式处理3T3-L1前脂肪细胞与成熟脂肪细胞,通过实时荧光定量PCR检测炎症因子,包括肿瘤坏死因子(tumor necrosis factor,TNF-ɑ)、白细胞介素6(interleukin 6,IL-6)与Ⅰ型干扰素(typeⅠinterferon,包括IFN-α和IFN-β)在mRNA水平的表达;另外通过ELISA的方法检测炎症因子与Ⅰ型干扰素在蛋白水平的表达变化.结果 与对照组相比,两种Poly(I:C)的处理方式都提高了炎症因子和Ⅰ型干扰素在mRNA水平的表达,但是转染的方式显著提高了炎症因子和Ⅰ型干扰素在mRNA水平的表达,同时在蛋白水平也发现转染方式显著促进炎症因子和Ⅰ型干扰素的分泌;在分化成熟的3T3-L1脂肪细胞中也发现了转染方式增强了Poly(I:C)的固有免疫应答.外源直接加入刺激了Toll样受体3(Toll-like receptor 3,TLR3)的表达,而转染方式刺激了TLR3、黑色素瘤分化相关蛋白5(mel-anoma differentiation related proteins 5,MDA5)和维甲酸诱导基因Ⅰ(retinoic acid induced gene I,RIG-Ⅰ)受体的表达.结论 通过对于Poly(I:C)的不同处理方式刺激脂肪细胞产生的免疫应答的研究,发现转染方式相比较直接处理方式,显著提高了Poly(I:C)引发的炎症反应.

abstractsObjective To investigate the innate immune responses in preadipocytes and adipocytes induced by polyinosinic polycytidylic acid [ Poly ( I : C ) ] with different stimulation strategies . Methods Preadipocytes (3T3-L1) and mature adipocytes were treated with Poly(I : C) through direct exogenous addi-tion and endogenous transfection. Expression of inflammatory factors (TNF-ɑ and IL-6) and typeⅠinterfer-ons ( IFN-α and IFN-β) were detected by real-time quantitative PCR at mRNA level and by ELISA at pro-tein level. Results Compared with control group, both Poly( I : C) stimulation strategies improved the ex-pression of inflammatory factors and typeⅠinterferons at mRNA and protein levels. Transfection of Poly(I : C) to preadipocytes (3T3-L1) and mature adipocytes significantly enhanced the expression of in-flammatory factors and typeⅠinterferons at mRNA and protein levels as compared with the strategy of direct exogenous addition. Exogenous addition of Poly ( I : C ) induced the expression of Toll-like receptor 3 (TLR3), while transfection of Poly(I : C) induced the expression of TLR3, melanoma differentiation relat-ed proteins 5 ( MDA5 ) and retinoic acid inducedⅠreceptor ( RIG-Ⅰ) . Conclusion This study suggests that transfection of Poly( I : C) into preadipocytes and adipocytes could induce more inflammation reactions than directly addition.