益气温阳方对变应性鼻炎Treg细胞FOXP3基因甲基化的影响  

Effect of Yiqi Wenyang Decoction on FOXP3 Gene Methylation in Treg Cells of Allergic Rhinitis

作　　者：刘玉[1] 史军[2] LIU Yu;SHI Jun(Nanjing Integrated Traditional Chinese and Western Medicine Hospital Affiliated of Nanjing University of Chinese Medicine,Nanjing 210014,Jiangsu,China;JiangsuProvincial Hospital of Traditional Chinese Medicine,Nanjing 210029,Jiangsu,China)

机构地区：[1]南京市中西医结合医院,江苏南京210014 [2]江苏省中医院,江苏南京210029

出　　处：《辽宁中医药大学学报》2020年第12期70-74,共5页Journal of Liaoning University of Traditional Chinese Medicine

基　　金：国家自然科学基金(81704126,81674035);江苏省自然科学基金(BK20171515);江苏省中医药科技项目(YB2017043);南京市科技发展计划项目(201803038)。

摘　　要：目的观察益气温阳方治疗变应性鼻炎(allergic rhinitis,AR)近期疗效及叉状头转录因子p3(forkhead boxp3,FOXP3)甲基化对益气温阳方治疗AR的免疫耐受机制的调控作用。方法60例AR患者在知情同意原则基础上按照随机数字表法分为中药组(益气温阳方)和西药组(氯雷他定)各30例,分别于治疗前后观察病情变化情况,采集血标本,用Western blotting检测FOXP3蛋白和DNA甲基转移酶(DNA methyltransferases,DNMT),用亚硫酸氢钠测序法(BSP)检测FOXP3基因甲基化水平及位点。结果中药组与西药组疗效比较,中药组优于西药组(P<0.05);AR患者血中的DNMT1、DNMT3a和DNMT3b浓度明显高于正常人水平(P<0.0001),而FOXP3值明显低于正常人水平(P<0.0001)。益气温阳方治疗后DNMT1、DNMT3a和DNMT3b浓度明显下降(P<0.0001)至正常组水平,而FOXP3值明显回升(P<0.0001),差异具有显著的统计学意义。用BSP法经FOXP3基因12个CpG位点的筛选表明:AR患者FOXP3基因GCP位点甲基化率明显高于正常人水平,二者差异有统计学意义(P<0.0001),益气温阳方治疗后FOXP3基因GCP位点甲基化率下降(P<0.05)。结论益气温阳方短期疗效优于氯雷他定;益气温阳方通过抑制DNMT,减少FOXP3基因GCP位点甲基化率,从而增加FOXP3基因表达,增加FOXP3蛋白表达,从而增加Treg细胞数量,进而抑制其后的一系列免疫反应,缓解AR症状。Objective To observe the short-term therapeutic effect of Yiqi Wenyang Decoction on allergic rhinitis(AR),the regulatory effect of FOXP3 methylation on immune tolerance mechanism of Yiqi Wenyang Decoction in the treatment of AR. Methods Based on the principle of informed consent,60 patients were divided into Chinese medicine group(Yiqi Wenyang Decoction)and Western medicine group(loratadine)according to the random number table method,30 cases each. Before and after the treatment,the changes of disease conditions were observed,blood samples were collected,Western blotting was used to detect FOXP3 protein and DNMT,and sodium bisulfite sequencing(BSP)was used to detect the methylation level and site of FOXP3 gene. Results(1)Compared with the Western medicine group,the traditional Chinese medicine group was superior to the western medicine group(P<0.05);(2)The concentration of DNMT1,Dnmt3 a and DNMT3 b in the blood of AR patients was significantly higher than that of normal people(P<0.000 1),while the value of FOXP3 was significantly lower than that of normal people(P<0.000 1). After treatment of Yiqi Wenyang Decoction,the concentrations of DNMT1,DNMT3 a and DNMT3 b decreased significantly(P<0.000 1)to the normal group,while the FOXP3 value increased significantly(P<0.000 1),which was statistically significant. The BSP method was used to screen 12 CpG sites of FOXP3 gene. The results showed that the methylation rate of GCP site of FOXP3 gene in AR patients was significantly higher than that in normal people(P<0.000 1). After treatment with Yiqi Wenyang Decoction,the methylation rate of GCP site of FOXP3 gene decreased(P<0.05). Conclusion(1)The short-term effect of Yiqi Wenyang Decoction is better than that of loratadine;(2)Yiqi Wenyang Decoction can reduce the methylation rate of FOXP3 gene GCP site by inhibiting DNMT,thus increasing FOXP3 gene expression and FOXP3 protein expression,so as to increase the number of Treg cells,thus inhibiting a series of subsequent immune responses and alleviating AR symptoms.

关 键 词：益气温阳方 变应性鼻炎 FOXP3基因甲基化 TREG细胞 中医药治疗 

分 类 号：R765.21[医药卫生—耳鼻咽喉科]