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目的:观察PolyI:C对大鼠气道平滑肌细胞(ASMCs)Toll样受体3(TLR3)及炎性细胞因子表达的影响,探讨ASMCs内TLR3表达与气道炎症间的关系。方法:体外培养大鼠ASMCs,传代培养后分为正常对照组和PolyI:C刺激组,PolyI:C刺激组又分为不同的时间点。用RT-PCR法检测细胞TLR3mRNA的表达;Western blot法检测ASMCs中核因子κB(NF-κB)的蛋白含量;ELISA法检测细胞培养上清Eotaxin和IL-8的浓度。结果:与正常对照组相比较,PolyI:C刺激组ASMCs内TLR3mRNA和NF-κB蛋白表达增加(P<0.05);ASMCs培养上清液中IL-8和Eotaxin浓度增加(P<0.05),并存在时间依赖性。结论:PolyI:C可以上调ASMCs内TLR3的表达,活化NF-κB,增加趋化因子Eotaxin和IL-8的浓度,参与哮喘的气道炎症反应。
Objective: To observe the effect of poly I: C on the expression of Toll like receptor 3 (TLR3) and inflammatory cytokines in rat airway smooth muscle cells (ASMCs), and to explore the relationship between TLR3 expression and airway inflammation in ASMCs. Methods: Rat ASMCs were cultured in vitro and divided into normal control group and PolyI: C stimulation group after subculture. PolyI: C stimulation group was divided into different time points. The expression of TLR3 mRNA was detected by RT-PCR. The protein level of nuclear factor kappa B (NF-κB) was detected by Western blot and the concentration of Eotaxin and IL-8 by ELISA. Results: Compared with the normal control group, the expression of TLR3mRNA and NF-κB protein in ASMCs treated with PolyI: C increased (P <0.05), while the concentrations of IL-8 and Eotaxin in ASMCs increased (P <0.05) There is a time dependency. CONCLUSION: PolyI: C can up-regulate the expression of TLR3, activate NF-κB, increase the concentrations of chemotactic factors Eotaxin and IL-8 in ASMCs, and participate in airway inflammation in asthma.