【摘要】 目的研究白细胞介素10(IL-10)在伴放线放线杆菌(Aa)脂多糖(LPS)刺激兔肺泡巨噬细胞(AM)产生炎症反应过程中的免疫调节作用及其机制。方法分离纯化AM,100ng/mL IL-10预孵2h,加入Aa-LPS刺激继续培养24h,流式细胞术检测AM对荧光标记Aa(FITC-Aa)的吞噬指数(PI)情况及AM的活性氧(ROS)水平;荧光定量PCR法检测CD14、TLR4、SRA和MHC-Ⅱ基因表达的变化;硝酸还原酶法检测AM的NO分泌情况。结果 IL-10上调Aa-LPS诱导的AM的PI(P<0.05),下调Aa-LPS诱导的AM膜表面受体CD14、TLR4、MHC-ⅡmRNA的表达(P<0.01),而SRA mRNA表达量升高(P<0.05);IL-10抑制Aa-LPS诱导的AM的ROS和NO水平(P<0.01)。结论 IL-10可以通过上调Aa-LPS诱导的AM SRA基因表达,下调CD14、TLR4、MHC-Ⅱ基因表达,并且抑制其产生ROS和NO的水平,从而调节免疫反应,限制炎症介质的大量释放,同时增强AM的吞噬作用,清除感染。更多还原

【Abstract】 Objective To evaluate the potential immunomodulatory effects and explore the possible mechanisms of interlukin 10on activation of Actinobacillus actinomycetemcomitans(Aa)-LPS induced alveolar macrophages(AM).Methods Isolation and purification of AM,AM was pre-incubated with 100 ng/mL IL-10for 2hand then activated by Aa-LPS for 24h:phagocytic index(PI)was measured by flow cytometry;RT-PCR was used to detect the expression of gene CD14,TLR4,SRA,MHC-Ⅱ;flow cytometry was used to assess ROS;nitrate reductase assay was used to examin the amount of NO production.Results IL-10increase the PI of Aa-LPS activated AM(P<0.05),remarkably inhibited the expressions of CD14,TLR4,MHC-Ⅱ(P<0.01),but enhanced the expression of SRA(P<0.05),significantly suppressed the ROS(P<0.01)and NO production(P<0.01).Conclusions IL-10can modulate of immune response and down-regulate excessive inflammatory mediator production and enhance phagocytosis of AaLPS activated AM via increasing SRA,suppressing CD14,TLR4,MHC-Ⅱexpression,and inhibit ROS,NO production.更多还原