摘要
目的通过检测特定性皮炎(AD)模型小鼠皮损中Toll样受体(TLR)-4和核转录因子(NF)-κB p65表达,探寻龙牡汤治疗AD的作用的可能机制。方法取18只BALB/c小鼠,将其中12只制作成AD模型小鼠,并将其分为3组(中药组、模型组、空白组),通过对比3组小鼠的血清中白细胞介素(IL)-4、IgE、干扰素(IFN)-γ以及局部皮损中TLR-4、NF-κB p65的差异,分析其相关性,探讨龙牡汤发挥作用的机理。结果 (1)TLR-4、NF-κB p65在3组中表达差异有统计学意义,其中模型组最高,中药组其次,空白组最小(P<0.05);(2)TLR-4和NF-κB p65的表达呈正相关(r=0.618,P<0.05);(3) IL-4、IgE、IFN-γ与TLR-4/NF-κB p65通路相关性分析:IL-4与TLR-4、NF-κB p65呈正相关,相关系数分别是:0.485、0.569(P<0.05);IgE与TLR-4、NF-κB p65呈正相关,相关系数分别是:0.651、0.721(P<0.05);IFN-γ与TLR-4、NF-κB p65呈正相关,相关系数分别是:0.55、0.634(P<0.05)。结论龙牡汤有可能是通过影响TLR-4表达进而阻断NF-κB p65通路释放炎症因子,从而平衡Th1/Th2细胞亚群来发挥免疫调节作用。
Objective In order to explore the possible mechanism of Longmu decoction in treating atopic dermatitis(AD) by detecting the expression of TLR-4,NF-κB p65 in the skin lesions of AD mice models. Methods Eighteen BALB/c mice were selected and 12 of them were used as AD mice models. AD mice were divided into three groups(traditional Chinese medicine group,model group and blank group). By comparing the expression levels of IL-4,IgE,IFN-γ in serum and TLR-4,NF-κB p65 in local skin lesions of three groups of mice, the correlation was analyzed,and exploring the therapeutic mechanism of Longmu decoction. Results LSD statistical analysis showed that the expressing levels of TLR-4,NF-κB p65 in the three groups were different(P<0.05),model group was the highest,traditional Chinese medicine group was followed,the blank group was minimum. The expressing levels of TLR-4 and NF-κB p65 protein in lesions of AD mice were analyzed by Spearman correlation. It was found that the expression of TLR-4,NF-κB p65 were positively correlated(r=0.618,P<0.05).We analyzed the correlation between serum inflammatory factors(IL-4,IgE,-γ) and TLR-4/NF-κB p65 pathway,results of statistical analysis showed IL-4 was positively correlated with TLR-4,NF-κB p65,with correlation coefficients of 0.485 and 0.569(P<0.05),IgE was positively correlated with TLR-4,NF-κB p65,with correlation coefficients of 0.651 and 0.721(P<0.05),IFN-γ was positively correlated with TLR-4 and NF-κB p65,with correlation coefficients of 0.55 and 0.634(P <0.05). Conclusion Longmu decoction may play an immunoregulatory role by affecting the expression of TLR-4 and then blocking the NF-κB p65 pathway to release inflammatory factors,thereby balancing Th1/Th2 cell subsets.
引文
[1]迟慧彦.特应性皮炎中医药治疗临床疗效评价——镇心安神法治疗特应性皮炎的随机对照研究[D].北京:中国中医科学院,2012.
[2]莫俊銮,周继昌,刘小立,等.DNCB致敏诱导BALB/c小鼠特应性皮炎模型的建立[J].中国热带医学,2013,13(12):1 439-1 442.
[3]Bieber T.Atopic dermatitis[J].Ann Dermatol,2010,22:125-137.
[4]李炜,王小波.Toll样受体与特应性皮炎[J].全科医学临床与教育,2012,10(4):407-410.
[5]Akira S,Hemmi H.Recognition of pathogen-associated molecular patterns by TLR family[J].Immunol Lett,2003,85:85-95.
[6]Wullaert A,Bonnet MC,Pasparakis M.NF-κB in the regulation of epithelial homeostasis and inflammation[J].Cell Res,2011,21:146-158.
[7]Carmody RJ,Chen YH.Nuclear factor-kappa B:activation and regulation during toll-like receptor signaling[J].Cell Mol Immunol,2007,4:31-41.
[8]Kim YJ,Jang ER,Lee JC,et al.Diarylheptanoid hirsutenoxime inhibits toll-like receptor 4-mediated NF-κB activation regulated by Akt pathway in keratinocytes[J].Am J Chin Med,2010,38:1 207-1 222.
[9]Mempel M,Voelcker V,Kollisch G,et al.Toll-like receptor expression in human keratinocytes:nuclear factorκB controlled gene activation by Staphylococcus aureus is toll-like receptor 2but not toll-like receptor 4 or platelet activating factor receptor dependent[J].J Invest Dermatol,2003,121:1 389-1 396.