苦参通过抑制MAPK/NF-κB通路改善免疫平衡缓解特应性皮炎实验研究  

Sophora Flavescens Alleviates Atopic Dermatitis by Improving Immune Balance and Inhibiting MAPK/NF-κB Signaling Pathways

作　　者：黄盼[1] 潘意 王畅[1] 汪海珍[1] 胡凡[2] HUANG Pan;PAN Yi;WANG Chang;WANG Hai-zhen;HU Fan(Department of Dermatology,Second Affiliated Hospital,Hunan University of Chinese Medicine,Changsha,410005;Department of Anorectal Disease,Second Affiliated Hospital,Hunan University of Chinese Medicine,Changsha,410005)

机构地区：[1]湖南中医药大学第二附属医院皮肤科,长沙410005 [2]湖南中医药大学第二附属医院肛肠科,长沙410005

出　　处：《中国中西医结合杂志》2024年第11期1365-1371,共7页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：湖南省自然科学基金青年基金资助项目(No.2023JJ40493)。

摘　　要：目的 探讨苦参对特应性皮炎(AD)辅助型T细胞1/辅助型T细胞2(Th1/Th2)失衡的治疗作用及其内在分子机制。方法 将24只雌性C57BL/6小鼠,按体重分层随机分为正常对照组、AD模型组和AD+苦参组,每组8只。采用2,4-二硝基氯苯/粉尘螨提取物(DNCB/DFE)诱导建立AD小鼠模型。将人表皮角质形成细胞(HaCaT)分成对照组、干扰素-γ(IFN-γ)/肿瘤坏死因子-α(TNF-α)组、IFN-γ/TNF-α+苦参组,共3组。采用实时荧光定量聚合酶链式反应(q RT-PCR)检测IFN-γ、TNF-α、白细胞介素(IL)-2、胸腺激活调节趋化因子(TARC)、IL-4、IL-5、IL-6 mRNA表达,Western Blot检测小鼠组织和细胞中磷酸化有丝分裂原活化蛋白激酶(p-p38)、p38、磷酸化核因子κB p65亚单位(p-p65)、p65蛋白表达,ELISA检测IL-6、TARC释放水平以及流式Th1/Th2细胞比例。结果 在细胞水平方面:与对照组比较,IFN-γ/TNF-α组IL-6、TARC mRNA及蛋白表达水平显著升高(P<0.01),p-p38、p-p65蛋白表达均上调(P<0.01);与IFN-γ/TNF-α组比较,IFN-γ/TNF-α+苦参组IL-6、TARC mRNA和蛋白表达水平均显著降低(P<0.01),p-p38、p-p65蛋白表达显著下调(P<0.01)。在动物水平研究中:与正常对照组比较,AD模型组小鼠皮肤组织中p-p38、p-p65蛋白表达水平显著增加(P<0.01);耳部皮肤组织炎症细胞浸润增多,角质化过度、肥大细胞浸润增加(P<0.01),且皮肤中Th2细胞含量增加(P<0.01);小鼠皮肤中IFN-γ、TNF-α、TARC、IL-6、IL-4、IL-5表达增加(P<0.01);与AD模型组比较,AD+苦参治疗组小鼠皮肤组织中p-p38、p-p65蛋白表达水平均减少(P<0.01);表皮、真皮厚度和肥大细胞数均显著降低(P<0.05);Th2细胞数量及组织中TARC、IL-6、IL-4、IL-5表达水平显著降低(P<0.01)。结论 苦参可通过抑制MAPK/NF-κB信号通路改善Th1/Th2免疫平衡从而减轻AD的症状。Objective To study the therapeutic effects of Sophora flavescens on T helper 1 cell/T helper 2 cell(Th1/Th2)imbalance in atopic dermatitis(AD)and its potential mechanism.Methods Totally 24 female C57BL/6 mice were randomly divided into normal control group,AD group,and AD+Sophora flavescens group by stratified body weight,8 mice in each group.AD mouse model was induced by 2,4-dinitrochlorobenzene/Dermatophagoides farinae extract(DNCB/DFE).HaCaT were divided into three groups:control group,interferon-γ( IFN-γ)/tumor necrosis factor-α( TNF-α) group, and IFN-γ/TNF-α+Sophora flavescens group.Quantitative Real-time polymerase chain reaction (qRT-PCR) was used to detect mRNA expressions of IFN-γ,TNF-α, interleukin-2 (IL-2), thymus activation-regulated chemokine (TARC), IL-4, IL-5, and IL-6. The proteinexpressions of phospho-mitogen activated protein kinase p38 (p-p38), p38, phospho-nuclear factor kappa B p65subunit (p-p65) and p65 in tissues and cells were detected by Western Blot. ELISA was used to detect the releaselevels of IL-6 and TARC, and the ratio of Th1/Th2 cells in flow cytometry. Results At the cell level: Comparedwith the control group,IL-6 and TARC mRNA and protein expressions significantly increased (P<0.01), andexpressions of p-p38 and p-p65 proteins were up-regulated (P<0.01) in IFN-γ/TNF-α group. Compared withthe IFN-γ/TNF-αgroup,IL-6 and TARC mRNA and protein expressions significantly decreased(P<0.01), andthe protein expressions of p-p38 and p-p65 were significantly down-regulated (P<0.01) in the IFN-γ/TNF-α+Sophora flavescens group. At the animal level, compared with the normal control group, the expression levelsof p-p38 and p-p65 protein in the skin tissue significantly increased (P<0.01), infiltration of inflammatory cellsincreased in the ear skin tissue, hyperkeratinization and infiltration of mast cells also increased (P<0.05), andTh2 cells increased in the skin (P<0.01), expressions of IFN-γ, TNF-α, TARC, IL-6, IL-4, and IL-5 in mouseskin increased (P<0.01) in the AD model grou

关 键 词：苦参 特应性皮炎 辅助型T细胞1/辅助型T细胞2失衡 有丝分裂原活化蛋白激酶/核因子κB信号通路 中药 

分 类 号：R285.5[医药卫生—中药学]