基于DNA甲基化探究祛风消癜方治疗过敏性紫癜风热伤络证的作用机制

Based on DNA methylation to explore the mechanism of QuFengXiaoDian Formula in the treatment of Henoch-Schonlein Purpura with the Syndrome of wind-heat injuring col⁃laterals

目的探究祛风消癜方在DNA甲基化层面治疗过敏性紫癜(Henoch-Schonlein purpura,HSP)风热伤络证的作用机制。方法选取2018年5月至2019年12月河南中医药大学第一附属医院初发HSP风热伤络证患儿5例、健康志愿者5例,分别采集患儿治疗前后和健康儿的空腹静脉血,应用Illumina850K基因芯片对其进行检测,筛选甲基化差异位点、差异基因并对其进行分析。结果风热伤络证患儿经祛风消癜方治疗前后共注释到90个差异基因,其中63个基因发生甲基化水平上调、27个基因甲基化水平下调。KEGG Pathway富集分析:差异基因富集于EGFR酪氨酸激酶抑制剂耐药性、药物代谢-细胞色素P450、RNA转运、5-羟色胺能突触、FoxO信号通路等,其中FoxO信号通路富集的差异基因为BRAF、IL7R。结论祛风消癜方可能通过调节FoxO信号通路,使IL7R基因表达上调、BRAF表达下调,双重调控异常的高甲基化和低甲基化状态,影响机体细胞分化/凋亡、白细胞活化与免疫应答等过程,抑制Th17/Treg细胞免疫失衡以发挥其对HSP风热伤络证的干预作用。

Objective To explore the mechanism of QuTengXiaoDian Formula(QFXDF)on DNA methylation in the treatment of Henoch-Schönlein Purpura(HSP)with wind-heat injuring collaterals.Methods From May 2018 to December 2019,5 chil⁃dren with HSP wind-heat collateral injury syndrome and 5 healthy volunteers were selected from the first affiliated Hospital of Henan University of Chinese Medicine.Fasting venous blood was collected before and after treatment and healthy children were detected by Illumina 850K gene chip.Methylation difference sites and differential genes were screened and analyzed.Results A total of 90 differential genes were annotated in children with wind-heat injury syndrome before and after treatment,of which 63 genes were up-regulated and 27 genes were down-regulated.KEGG Pathway enrichment analysis:the differential genes were enriched in drug resistance of EGFR tyrosine kinase inhibitors,drug metabolism-cytochrome P450,RNA transport,serotonergic synapses,FoxO signal pathway and so on.The differential genes enriched in FoxO signal pathway were BRAF and IL7R.Conclu⁃sion QFXDF may regulate the FoxO signal pathway,up-regulate the expression of IL7R gene and down-regulate the expression of BRAF,double regulate the abnormal hypermethylation and hypomethylation,affect the process of cell differentiation/apopto⁃sis,leukocyte activation and immune response,and inhibit the immune imbalance of Th17/Treg cells to exert its intervention effect on the syndrome of wind-heat damaging collaterals in HSP.