柴朴汤对哮喘小鼠支气管上皮细胞炎症因子及HMGB1/TLR4/NF-κB的影响

The influence of Chaipu decoction on inflammatory factors and HMGB1/TLR4/NF-κ B in bronchial epithelial cells of asthmatic mice

目的探讨柴朴汤对哮喘小鼠支气管上皮细胞高迁移率族蛋白B1(HMGB1)、Toll样受体4(TLR4)、核因子-κB(NF-κB)及下游炎症介质单核细胞趋化蛋白-1(MCP-1)、白细胞介素-6(IL-6)的影响。方法将小鼠支气管上皮细胞分为对照组(Con组,空白血清培养)、Model组(哮喘小鼠血清培养)和Chaipu组(柴朴汤药血清培养)。采用qRT-PCR和Western blotting检测小鼠支气管上皮细胞中HMGB1、TLR4、NF-κB、MCP-1、IL-6的mRNA及蛋白表达情况,ELISA法测定细胞培养液中MCP-1和IL-6的水平。结果与Con组相比,Model组小鼠支气管上皮细胞HMGB1、TLR4、NF-κB、MCP-1、IL-6表达均明显升高(P<0.05);Model组细胞培养液中MCP-1、IL-6也显著升高(P<0.05)。与Model组相比,Chaipu组小鼠支气管上皮细胞HMGB1、TLR4、MCP-1、IL-6 mRNA和蛋白表达降低,NF-κB m RNA表达亦降低(P<0.05);Chaipu组细胞培养液中MCP-1、IL-6也显著降低(P<0.05)。结论柴朴汤可下调哮喘血清诱导的炎症因子的表达,可能与其抑制HMGB1/TLR4/NF-κB炎症通路有关。

Aim To investigate the effects of Chaipu decoction on high mobility group protein B1(HMGB1),Toll-like receptor 4(TLR4),nuclear factor-κB(NF-κB),and downstream inflammatory mediators monocyte chemotactic protein-1(MCP-1)and interleukin-6(IL-6)in bronchial epithelial cells of asthmatic mice.Methods The mouse bronchial epithelial cells were divided into a control group(Con group,treated with blank serum culture),a Model group(treated with asthma tic mice serum culture),and a Chaipu group(treated with Chaipu decoction containing serum culture).qRT-PCR and Western blotting were used to detect the mRNA and protein expression of HMGB1,TLR4,NF-κB,MCP-1,and IL-6 in mouse bronchial epithelial cells,and ELISA assay was used to determine the levels of MCP-1 and IL-6 in the cell culture fluid.Results Compared with the Con group,the expression of HMGB1,TLR4,NF-κB,MCP-1,and IL-6 in mouse bronchial epithelial cells in the Model group were significantly increased(P<0.05).The MCP-1 and IL-6 of the cell culture fluid in the Model group were also significantly increased(P<0.05).The mRNA and protein expression of HMGB1,TLR4,MCP-1,IL-6 were decreased and the expression of NF-κB mRNA was decreased in the Chaipu group compared with the Model group(P<0.05).The levels of MCP-1 and IL-6 in Chaipu group cell culture medium were also significantly decreased(P<0.05).Conclusion Chaipu decoction can downregulate the expression of inflammatory factors induced by asthma serum,which may be related to its inhibition of HMGB1/TLR4/NF-κB inflammatory pathway.