龙牡汤对特应性皮炎小鼠树突状细胞IgE高亲和力受体γ亚基启动子甲基化水平及Th分化的影响  

Effects of Longmu Decoction on methylation level of high-affinity IgE receptorγsubunit promoter and Th differentiation in dendritic cells of mice with atopic dermatitis

作　　者：宋艳丽[1] 刘青云[1] 陈少君[1] 佘远遥[1] 王晶晶[1] 姚春海[1] SONG Yan-li;LIU Qing-yun;CHEN Shao-jun;SHE Yuan-yao;WANG Jing-jing;YAO Chun-hai(Department of Dermatology,Xiyuan Hospital,China Academy of Chinese Medical sciences,Beijing 100091,China)

机构地区：[1]中国中医科学院西苑医院皮肤科,北京100091

出　　处：《北京中医药》2022年第4期379-384,共6页Beijing Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金青年基金项目(81503447)。

摘　　要：目的探讨自拟方龙牡汤对特应性皮炎(AD)小鼠树突状细胞(BMDC)IgE高亲和力受体(high affinity IgE receptor,FcERI)γ亚基启动子甲基化水平以及对Th2型免疫反应的影响。方法将30只BALB/c小鼠分成模型组、中药组、空白组,各10只。将模型组、中药组共20只小鼠制作AD模型,中药组给予龙牡汤灌胃,模型组、空白组给予浓度0.9%生理盐水灌胃,均1次/d,连续灌胃3周。将各组小鼠常规脱颈椎处死,取股骨和胫骨,分离并培养BMDC;中药组、模型组小鼠处死后,立刻经腹主动脉采血,分离血清,分别配成10%含龙牡汤血清和10%非含药血清。将BMDC随机分为对照组与给药组,对照组加入10%非含药血清,给药组加入10%含龙牡汤血清继续培养48 h。用流式细胞术检测BMDC的CD11c阳性率,用Western blotting法检测BMDC细胞FCER1γ蛋白表达,甲基化特异性PCR(MSP)检测,ELISA法检测BMDC细胞培养液上清中MCP-1、IL-12水平;流式细胞术检测BMDC细胞培养液CD4、IL-4双阳性率。结果与对照组比较,中药组BMDC细胞FCER1γ蛋白表达低(P<0.05);含药血清组FCER1γ亚基启动子甲基化水平高于对照组(P<0.05);中药组比对照组MPC-1水平低,IL-12水平高(P<0.05);与空白组比较,中药组的CD4和IL-4双阳性率差异无统计学意义(P>0.05)。结论龙牡汤能够提高AD小鼠突状细胞FcERⅠγ亚基基因调节序列甲基化水平,进而抑制FcERⅠγ亚基与FcERⅠ在细胞表面过度表达,促进了Th1分化,抑制了Th2型免疫反应。Objective To investigate the effect of self-designed formula Longmu Decoction on the methylation level of high-affinity IgE receptor(FcERI)γsubunit promoter and Th2 immune response in mice with atopic dermatitis(AD).Methods Animal grouping and modeling:30 BALB/C mice were divided into model group,traditional Chinese medicine group and blank group,with10 mice in each group.Among them,20 mice in model group and traditional Chinese medicine group were used to make AD model.Traditional Chinese medicine group was given Longmu Decoction by intragastric administration,and the model group and the blank group were given 0.9%normal saline by intragastric administration.All mice were given intragastric administration once a day for 3weeks.First,the mice in each group were routinely sacrificed by cervical dislocation,femur and tibia of them were obtained,and bone marrow dendritic cells(BMDC)were isolated and cultured;then,the blood of Chinese medicine group and model group mice were immediately collected through the abdominal aorta,the serum was separated,and 10%serum containing Longmu decoction and10%non-medicated serum were prepared respectively.BMDC was randomly divided into the control group and medicine group.The control group was added with 10%non-medicated serum and the medicine group was added with 10%serum containing Longmu Decoction both continuously cultured for 48 hours.CD11c positive rate of BMDC was detected by flow cytometry.The expression of FCER1γprotein in BMDC cells was detected by Western blotting.Methylation specific PCR(MSP)detection;the levels of McP-1and IL-12 in the supernatant of BMDC cells were determined by ELISA.The double positive rate of CD4 and IL-4 in BMDC was detected by flow cytometry.Results Compared with the control group,the expression of FCER1γprotein in BMDC was lower in Chinese medicine group(P<0.05),and the methylation level of FCER1γsubunit promoter in the drug-containing serum group was higher than that in the control group(P<0.05).Compared with control group,the level of MPC-1

关 键 词：特应性皮炎 龙牡汤 小鼠 树突状细胞 IgE高亲和力受体γ亚基 甲基化 Th2型免疫反应 

分 类 号：R285.5[医药卫生—中药学]