过敏煎通过IgE/FcεRⅠ途径抑制肥大细胞脱颗粒改善特应性皮炎的作用机制  

Mechanism of Guomin Decoction(过敏煎)to Improve Atopic Dermatitis by Inhibiting Mast Cell Degranulosis through IgE/FcεRI Pathway

作　　者：高娟 陈桂芳 谭伟 张洁 饶高雄 王睿睿 张祎 GAO Juan;CHEN Guifang;TAN Wei;ZHANG Jie;RAO Gaoxiong;WANG Ruirui;ZHANG Yi(School of Traditional Chinese Medicine,Yunnan University of Chinese Medicine,Kunming 650500,Yunnan,China;Yunnan Province Engineering Laboratory For National Healthcare Theories and Products,Kunming 650500,Yunnan,China;Yunnan Medical Health College,Kunming 650500,Yunnan,China)

机构地区：[1]云南中医药大学中药学院,云南昆明650500 [2]云南省民族特色养生理论与健康产品工程实验室,云南昆明650500 [3]云南医药健康职业学院,云南昆明650500

出　　处：《中华中医药学刊》2024年第9期135-140,I0025,I0026,共8页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(82160782,81760741);云南省科技厅联合专项[2017FF117(-025),2019FF002(-051)]。

摘　　要：目的通过体内外实验,研究过敏煎改善特应性皮炎的作用机制。方法采用2,4-二硝基氯苯(2,4-dimethyl sulfoxide,DNCB)反复刺激皮肤建立特应性皮炎(Atopic dermatitis,AD)模型,给予氯雷他定片及过敏煎低、中、高剂量(1.32、2.64、5.28 g/kg)进行干预,ELISA法检测血清总IgE、IL-4、IL-13水平,IHC法测定皮损组织中IL-4、IL-13的表达水平。体外运用anti-DNP-IgE/DNP-HSA诱导RBL-2H3细胞复制脱颗粒模型,MTS法检测过敏煎对RBL-2H3活力,通过ELISA法检测细胞上清中β-hex酶释放率,Histamine、LTB4、IL-4、IL-13水平,RT-PCR法检测细胞Histamine、IL-4、IL-13mRNA表达,Western blotting法检测p-ERK/ERK、p-JNK/JNK、p-p38/p-38蛋白表达。结果与模型组比较,过敏煎各剂量组显著减轻AD小鼠血清总IgE、IL-4、IL-13水平(P<0.05,P<0.01),减轻AD小鼠皮损组织中IL-4、IL-13表达。明显下调RBL-2H3细胞脱颗粒后β-Hex酶释放率(P<0.05,P<0.01)和Histamine、LTB4、IL-4、IL-13水平(P<0.05,P<0.01),显著减少细胞Histamine、IL-4、IL-13mRNA表达(P<0.01)和p-ERK/ERK、p-JNK/JNK、p-p38/p38蛋白表达(P<0.05,P<0.01)。结论过敏煎通过调控Th2型反应和血清中总IgE水平改善特应性皮炎,其作用机制可能是通过IgE/FcεRⅠ途径抑制肥大细胞脱颗粒。Objective To study the mechanism of Guomin Decoction(过敏煎)in improving atopic dermatitis through in vitro and in vivo experiments.Methods The model of atopic dermatitis(AD)was established by repeated stimulation of 2,4-dinitro-chlorobenzene(DNCB).Loratadine Tablets and Guomin Decoction with low,medium and high doses(1.32,2.64,5.28 g/kg)were administered.The levels of total immunoglobulin E(IgE),interleukin-4(IL-4)and interleukin-13(IL-13)in serum were detected by ELISA.The expression levels of IL-4 and IL-13 in skin lesions were determined by immunohistochemistry(IHC)method.In vitro,anti-DNP-IgE/DNP-HSA was used to induce RBL-2H3 cell replication degranulation model.MTS method was used to detect the activity of Guomin Decoction on RBL-2H3.ELISA was used to detect theβ-Hex enzyme release rate and the levels of histamine,leukotriene B4(LTB4),IL-4 and IL-13 in cell supernatant.The mRNA expressions of histamine,IL-4 and IL-13 were detected by reverse transcription-polymerase chain reaction(RT-PCR),and the protein ex-pressions of phosphorylation-extracellular regulated protein kinases(p-ERK)/ERK),phosphorylation-c-Jun N-terminal kinase(p-JNK)/JNK and p-p38/p38 were detected by Western blotting.Results Compared with those of the model group,the levels of total IgE,IL-4 and IL-13 in serum of AD mice in Guomin Decoction groups were significantly decreased(P<0.05,P<001),and the expressions of IL-4 and IL-13 in skin lesions of AD mice were decreased.β-HEX enzyme release rates(P<0.05,P<001)and the levels of histamine,LTB4,IL-4 and IL-13(P<0.05,P<001)were significantly decreased after degranulation of RBL-2H3 cells.The mRNA expressions of histamine,IL-4 and IL-13(P<001)and protein expres-sions of p-ERK/ERK,p-JNK/JNK and p-p38/p38(P<0.05,P<001)were significantly decreased.Conclusion Guomin Decoction ameliorates atopic dermatitis by regulating Th2 type response and total IgE level in serum,possibly through IgE/FcεRI pathway inhibiting mast cell degranulosis.

关 键 词：过敏煎 特应性皮炎 IgE/FcεRⅠ 肥大细胞 脱颗粒 

分 类 号：R289.5[医药卫生—方剂学]