GEO数据库联合网络药理学及实验验证探讨薏竹温胆汤治疗湿疹作用机制  

Exploration on the Mechanism of Yizhu Wendan Decoction in Treating Eczema Based on GEO Database Combined with Network Pharmacology and Experimental Verification

作　　者：王钇杰 郭婷婷 李勇军 李孜怡 张萌[1,2] 史梦迪 谷胜男 王有鹏 李志军[1,2] WANG Yijie;GUO Tingting;LI Yongjun;LI Ziyi;ZHANG Meng;SHI Mengdi;GU Shengnan;WANG Youpeng;LI Zhijun(Heilongjiang University of Chinese Medicine,Harbin 150040,China;The Second Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150001,China;The First Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150040,China;Affiliated Hospital of Shanxi University of Chinese Medicine,Taiyuan 030024,China;Shenzhen Hospital,Beijing University of Chinese Medicine,Shenzhen 518116,China)

机构地区：[1]黑龙江中医药大学,黑龙江哈尔滨150040 [2]黑龙江中医药大学附属第二医院,黑龙江哈尔滨150001 [3]黑龙江中医药大学附属第一医院,黑龙江哈尔滨150040 [4]山西中医药大学附属医院,山西太原030024 [5]北京中医药大学深圳医院,广东深圳518116

出　　处：《中国中医药信息杂志》2025年第4期32-41,共10页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：黑龙江省中医药科研项目(GY2022-24);黑龙江省博士后创新科研项目(LBH-Z20098);黑龙江中医药大学研究生创新科研项目(2023yjscx010);深圳市龙岗区科技发展专项-医疗卫生科技计划项目(LGKCYLWS2020079)。

摘　　要：目的通过GEO数据库联合网络药理学方法探讨薏竹温胆汤治疗湿疹的作用机制,并进行实验验证。方法利用TCMSP、BATMAN-TCM和ETCM数据库筛选薏竹温胆汤的活性成分,通过GEO数据库收集湿疹相关的疾病靶点信息,将药物活性成分靶点与疾病靶点取交集,构建药物-成分-靶点网络和蛋白相互作用网络,使用DAVID数据库进行GO和KEGG通路富集分析。采用CCK-8法筛选薏竹温胆汤冻干粉最佳干预浓度。将HaCaT细胞分为对照组、模型组、薏竹温胆汤低浓度组、薏竹温胆汤高浓度组、si-IL-17RA组、si-IL-17RA+薏竹温胆汤低浓度组、si-IL-17RA+薏竹温胆汤高浓度组、地塞米松组、si-IL-17RA+地塞米松组,并予相应干预。qPCR检测湿疹相关趋化因子和炎症因子表达,EdU和AnnexinⅤ-FITC/PI双染色法分别检测细胞增殖和凋亡水平,Western blot检测与凋亡、皮肤屏障及IL-17信号通路相关的蛋白表达。结果通过数据库检索得到薏竹温胆汤活性成分180个,结合湿疹相关的GEO数据库微阵列(GSE6012、GSE57225),得到薏竹温胆汤治疗湿疹的潜在作用靶点8个,KEGG通路富集分析结果主要涉及IL-17信号通路、脂质与动脉粥样硬化、TNF信号通路、流体剪切应力与动脉粥样硬化等。薏竹温胆汤冻干粉浓度为100μg/mL时细胞活力最强,薏竹温胆汤可显著抑制湿疹相关趋化因子及炎症因子CCL17、CCL22、IL-1β、TNF-α、IL-6、IFN-γmRNA表达,升高IL-4 mRNA表达;促进HaCaT细胞增殖,升高Bcl-2蛋白表达,降低Bad、Cleaved Caspase-3蛋白表达,从而抑制HaCaT细胞凋亡;促进FLG、LOR蛋白表达,降低IL-17信号通路MMP9、MMP1、CCL2、FOSL1、IL-17RA蛋白表达。结论薏竹温胆汤通过多成分、多途径、多靶点治疗湿疹,可促进HaCaT细胞增殖并抑制其凋亡,恢复皮肤屏障,其作用机制可能与抑制IL-17信号通路激活有关。Objective To explore the mechanism of Yizhu Wendan Decoction in treating eczema through GEO database combined with network pharmacology and experimental verification.Methods TCMSP,BATMAN-TCM and ETCM databases were used to screen the active components of Yizhu Wendan Decoction.Disease target information related to eczema was collected through GEO database.The drug-component-target network and PPI network were constructed by intersections of active component targets and disease targets.GO and KEGG pathway enrichment analyses were performed using DAVID database.CCK-8 method was used to screen out the optimal intervention concentration of freeze-dried powder of Yizhu Wendan Decoction.HaCaT cells were divided into control group,model group,Yizhu Wendan Decoction low concentration group,Yizhu Wendan Decoction high concentration group,si-IL-17RA group,si-IL-17RA+Yizhu Wendan Decoction low concentration group,si-IL-17RA+Yizhu Wendan Decoction high concentration group,Dexamethasone group,si-IL-17RA+Dexamethasone group.Each group was given relevant intervention.The expressions of chemokines and inflammatory factors were detected by qPCR.EdU and Annexin V-FITC/PI double staining were used to detect cell proliferation and apoptosis.Western blot was performed to detect the expressions of proteins related to apoptosis,skin barrier and IL-17 signaling pathway.Results By using databases,180 active components of Yizhu Wendan Decoction were obtained.Combined with GEO database microarrays related to eczema(GSE6012 and GSE57225),8 potential targets of Yizhu Wendan Decoction in the treatment of eczema were obtained.KEGG enrichment pathway mainly involved IL-17 signaling pathway,lipid and atherosclerotic,TNF signaling pathway,fluid shear stress and atherosclerotic,etc.When Yizhu Wendan Decoction freeze-dried powder concentration was 100μg/mL,cell viability was the strongest.Yizhu Wendan Decoction could significantly inhibit the mRNA expressions of chemokines and inflammatory factors CCL17,CCL22,IL-1β,TNF-α,IL-6,IFNγ,and increase

关 键 词：湿疹 薏竹温胆汤 分消走泄 网络药理学 作用机制 细胞实验 

分 类 号：R285[医药卫生—中药学]