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目的观察脓毒症主要相关细胞因子肿瘤坏死因子α(TNFα)和干扰素γ(IFNγ)对巨噬细胞表面主要模式识别受体(PRRs)表达的影响。方法分离培养小鼠肺泡巨噬细胞,用TNFα、IFNγ(终浓度均为20ng/ml)分别刺激细胞3h、6h、12h,通过逆转录聚合酶链反应和免疫组织化学检测细胞内PRRs,包括白细胞分化抗原14(CD14)、Toll样受体4(TLR4)、清道夫受体(SR)、细菌脂蛋白受体TLR2和细菌DNA受体TLR9mRNA表达及其蛋白表达。结果TNFα和IFNγ表现为不同程度地上调与炎细胞激活作用有关的PRRs(CD14、TLR2、TLR9)(P<0.05),下调与炎细胞防御作用有关的SR(P<0.05),而对TLR4基因及蛋白表达无显著刺激作用(P>0.05)。结论效应细胞释放的促炎因子TNFα、IFNγ能从基因转录和蛋白水平上对细胞表面PRRs产生明显的反馈调控作用,对于促进失控性炎症反应的发生具有一定病理生理意义。
Objective To observe the effects of tumor necrosis factor α (TNFα) and interferon γ (IFNγ), the major cytokines related to sepsis, on the expression of major pattern recognition receptors (PRRs) on macrophages. Methods Mouse alveolar macrophages were isolated and cultured. The cells were stimulated with TNFα and IFNγ (final concentration of 20ng / ml) for 3h, 6h, 12h, respectively. The intracellular PRRs including leukocytes were detected by reverse transcription polymerase chain reaction and immunohistochemistry Differentiation antigen 14 (CD14), Toll-like receptor 4 (TLR4), scavenger receptor (SR), bacterial lipoprotein receptor TLR2 and bacterial DNA receptor TLR9mRNA expression and protein expression. Results TNFα and IFNγ showed upregulation of PRRs (CD14, TLR2 and TLR9) (P <0.05) and downregulation of inflammatory cells (P <0.05), but did not affect TLR4 gene And no significant stimulation of protein expression (P> 0.05). Conclusions The proinflammatory cytokines TNFα and IFNγ released from effector cells play an important role in feedback regulation of cell surface PRRs at gene transcription and protein level and have certain pathophysiological significance for promoting uncontrolled inflammatory response.