牛抵抗素对牛肺泡巨噬细胞TLR4/MyD88非依赖信号通路基因表达的影响

Analysis of differential expression of TLR4-MyD88 independent pathway genes under resistin-induced bovine alveolar macrophages

旨在探究牛抵抗素通过TLR4/MyD88非依赖信号通路诱发牛肺泡巨噬细胞的相关炎症反应机理。使用100 ng·mL^(-1)终浓度牛抵抗素诱导牛肺泡巨噬细胞,分别在0、1.5、3.0、6.0、12.0、24.0 h时采用qRT-PCR检测TLR4/MyD88非依赖信号通路相关基因(TLR4、TRAM、NF-κB)mRNA水平表达量,通过ELISA法检测下游IL-6、1L-1β、TNF-α等促炎细胞因子表达水平。结果显示,100 ng·mL^(-1)牛抵抗素处理牛肺泡巨噬细胞后,TLR4、TRAM、NF-κB基因表达量于6.0 h时极显著上调(P<0.01),诱导12.0 h时,相关基因表达量达到最高;IL-6、1L-1β、TNF-α等促炎细胞因子于1.5 h表达量极显著上调(P<0.01),且均存在时间依赖效应。抵抗素诱导牛肺泡巨噬细胞后,可激活TLR4/MyD88非依赖信号通路,通过信号传递,释放大量促炎细胞因子,引起肺部炎症。

In order to explore the mechanism of bovine resistin through TLR4-MyD88 independent signal pathway in bovine alveolar macrophages,bovine alveolar macrophages were exposed to resistin(100 ng·mL^-1)for 0,1.5,3.0,6.0,12.0,24.0 h,respectively.The relative mRNA expression of TLR4-MyD88 independent signal pathway genes(TLR4,TRAM,NF-κB)were detected by qRT-PCR,the expression of proinflammatory cytokine IL-6,1L-1βand TNF-αwere tested by ELISA methods.The results showed that resistin(100 ng·mL^-1)sharply upregulated the expression of TLR4-MyD88 independent signal pathway genes at 6.0 h(P<0.01).The expression of related genes reached the highest at 12.0 h.The expression of proinflammatory cytokines IL-6,IL-1βand TNF-αsignificantly increased at 1.5 h(P<0.01).Through the signal transmission,a large number of proinflammatory cytokines were released,causing pulmonary inflammation.