摘要
目的 通过网络药理学与动物实验探究芪玄抑甲宁治疗格雷夫斯病(GD)的作用机制。方法 应用中药系统药理学数据库与分析平台和化学专业数据库查筛芪玄抑甲宁的活性成分及其靶点,从Genecards、在线人类孟德尔遗传数据库等中查筛GD相关靶点;将药物与疾病共有靶点导入STRING平台构建蛋白质-蛋白质相互作用(PPI)网络并筛选核心靶点;使用Cytoscape 3.8.2软件绘制“药物-活性成分-靶点”网络图并筛选核心成分;应用Metascape平台对核心靶点进行富集分析。选取6周龄SPF级体重(20±2)g的雌性BALB/c小鼠40只,其中10只设为正常对照组,其余30只均用重组腺病毒Ad-TSHR-289免疫3次构建GD模型。将造模成功的小鼠采用随机数字表法分为GD模型组、甲巯咪唑组、芪玄抑甲宁组,每组8只。芪玄抑甲宁组灌胃芪玄抑甲宁50 g/kg,甲巯咪唑组灌胃甲巯咪唑片3.75 mg/kg,正常对照组、GD模型组灌胃同体积的饮用水,1次/d。连续给药4周后,采用酶联免疫吸附试验检测四组血清血管内皮生长因子(VEGF)含量,免疫组织化学染色法检测四组甲状腺组织血小板内皮细胞黏附分子-1(PECAM-1)的表达量。结果 共得到芪玄抑甲宁活性成分63种,靶点281个,GD靶点1 489个,药物与疾病共有靶点124个。PPI获得信号转导和转录激活因子3、丝氨酸/苏氨酸蛋白激酶1、VEGFA等21个核心靶点。药物-靶点分析得到木犀草素、山柰酚、槲皮素、熊果酸、β-谷甾醇等18种核心成分。GO分析获得炎症反应、血管新生、血管形态发生等954项,KEGG分析获得MAPK、PI3K/AKT、VEGF等92条通路。动物实验显示,GD模型组血清VEGF和甲状腺组织PECAM-1表达量高于正常对照组,且芪玄抑甲宁组低于GD模型组,差异有高度统计学意义(P<0.01)。结论 芪玄抑甲宁可能通过调控MAPK、PI3K/AKT、VEGF等信号通路产生抑制血管新生的作用治疗GD。
Objective To explore the mechanism of Qixuan Yijianing in the treatment of Graves disease(GD) based on network pharmacology and animal experiments. Methods The active constituents and targets of Qixuan Yijianing were detected by traditional Chinese medicine systems pharmacology database and analysis platform, and chemical database, and GD related targets were detected from Genecards, online human Mendelian genetic database, etc. The common targets of drugs and diseases were introduced into STRING platform to construct protein-protein interaction(PPI) network and screen core targets. Cytoscape 3.8.2 software was used to map the drug-active ingredient-target network and screen core components. The core target was enriched by Metascape platform. Forty 6-week-old female BALB/c mice with SPF grade and body weight of(20±2) g were selected, ten of which were set as normal control group, and the remaining 30 were immunized with recombinant adenovirus Ad-TSHR-289 for three times to construct GD model. The mice that modeled successfully were divided into GD model group, Methimazole group,and Qixuan Yijianing group by random number table method, with eight mice in each group. Qixuan Yijianing group was intragastric administration of Qixuan Yijianing 50 g/kg, Methimazole group was intragastric administration of Methimazole Tablets 3.75 mg/kg, normal control group and GD model group were intragastric administration of drinking water of the same volume, once a day. After four weeks of continuous administration, serum vascular endothelial growth factor(VEGF) content was detected by enzyme-linked immunosorbent assay. The expression of platelet endothelial cell adhesion molecule-1(PECAM-1) in thyroid tissues of the four groups was detected by immunohistochemistry. Results A total of 63 active ingredients with 281 targets and 1 489 targets of GD were obtained. There were 124 targets of drugs and diseases. PPI obtained 21 core targets of signal transduction and transcription activator 3, serine/threonine protein kinase 1,VEGFA, etc. Eighteen core components including luteolin, kaempferol, quercetin, ursolic acid, and β-sitosterol were obtained by drug-target analysis. 954 items of inflammatory response, angiogenesis, and vascular morphogenesis were obtained GO by analysis, and 92 pathways including MAPK, PI3K/AKT, and VEGF were obtained by KEGG analysis.Animal experiments showed that the expression levels of serum VEGF and thyroid PECAM-1 in GD model group were higher than those in normal control group, and those in Qixuan Yijianing group were lower than those in GD model group,the differences were highly statistically significant(P<0.01). Conclusion Qixuan Yijianing is able to treat GD by regulating MAPK, PI3K/AKT, VEGF, and other signaling pathways to inhibit angiogenesis.
作者
高长久
丁崧
卢芳
柳长凤
于栋华
刘树民
GAO Changjiu;DING Song;LU Fang;LIU Changfeng;YU Donghua;LIU Shumin(Graduate School,Heilongjiang University of Chinese Medicine,Heilongjiang Province,Harbin150040,China;Institute of Chinese Medicine,Heilongjiang University of Chinese Medicine,Heilongjiang Province,Harbin150040,China)
出处
《中国医药导报》
CAS
2023年第3期9-14,共6页
China Medical Herald
基金
国家自然科学基金青年科学基金项目(81302898)
国家自然科学基金面上项目(82074149)。
关键词
芪玄抑甲宁
格雷夫斯病
网络药理学
血管新生
作用机制
Qixuan Yijianing
Graves disease
Network pharmacology
Angiogenesis
Mechanism of action
