Objective: To explore the effects of suppression on the c-Kit gene expression by RNA interference on the production of inflammatory cytokines mediated by alveolar macrophage activated by mite allergen.Methods: Alveolar macrophages obtained through bronchoalveolar lavage.CD4+ T cells were separated from peripheral blood with methods of Ficoll-Hypaque density-gradients centrifugation and magnetic cell sorting system (MACS).CD4+ T cells were co-cultured with alveolar macrophages activated by the mite allergen.Using specific siRNA inhibited the expression of c-kit gene.Flow Cytometry and Weatern blot detected effects of gene silence and Enzyme-Linked Immunosorbent Assay measured the production of inflammatory cytokines in culture supernatants.Results: siRNA could be transferred into the alveolar macrophages effectively with the aid of transfection agent and significantly inhibited the expression of c-Kit protein, and decreased the production of inflammatory cytokines.Conclusion: Anti-c-kit-siRNA can reduce the production of inflammatory cytokines mediated by alveolar macrophages.%目的:通过RNA干扰的方法抑制c-kit基因的表达,观察其对由螨变应原活化的肺泡巨噬细胞所介导的炎性因子生成的影响.方法:通过支气管肺泡灌洗获取肺泡巨噬细胞,密度梯度离心法及免疫磁珠分离出血液CD4+T细胞.经螨变应原活化后的肺泡巨噬细胞与CD4+T细胞共培养.采用抗c-kit特异性siRNA抑制其表达,流式细胞术及Weatern blot检测沉默效果,酶联免疫吸附法测定培养上清炎性因子.结果:siRNA在转染剂的介导下可有效转入肺泡巨噬细胞内,明显抑制了c-kit蛋白的表达,同时减少了肺泡巨噬细胞介导的炎性因子的生成.结论:抗c-kit特异性siRNA可有效抑制肺泡巨噬细胞介导的炎性因子的生成.
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