摘要： 目的 探讨逍遥补肾方治疗桥本甲状腺炎（HT）的潜在作用机制。方法 通过网络药理学对逍遥补肾方的成分靶点及HT的相关疾病靶点取交集，构建交集靶点的蛋白质-蛋白质相互作用（PPI）网络、药物-成分-交集靶点网络，并筛选出核心成分及靶点。随机将65只大鼠分为正常组（10只），造模组（55只）。高碘水喂养大鼠，联合多次甲状腺球蛋白注射建立实验性自身免疫性甲状腺炎（EAT）大鼠模型，造模成功后随机分为模型组、硒酵母组和逍遥补肾组。硒酵母组给予硒酵母片水溶液［25.2mg/（kg·d）］ ，逍遥补肾组给予逍遥补肾方溶液［11.79 mg/（kg·d）］ ，正常组、模型组给予等体积的蒸馏水。连续灌胃8周后收集血清，采用酶联免疫吸附试验（ELISA）检测各组大鼠血清中白蛋白（ALB）、蛋白激酶B1 （AKT1）和肿瘤坏死因子-α （TNF-α）的含量。收集粪便，采用超高液相色谱-串联质谱法（UPLC-MS/MS）进行代谢组学检测。结果 共获得逍遥补肾方治疗HT的6个核心成分，包括槲皮素、山奈酚、β-谷甾醇、豆甾醇、异鼠李素和木犀草素；5个核心靶点，包括ALB、AKT1、TNF、白细胞介素-6 （IL-6）和TP53。动物实验结果显示，与硒酵母组比较，逍遥补肾组ALB、AKT1、TNF-α的表达明显下调（P < 0.01）。代谢组学分析结果显示共鉴定出5个共同差异代谢物，包括（4Z，7Z，10Z，13Z，16Z，19Z）-二十二碳六烯酸乙酯、（S）-（+）-柠苹酸、2-酮-6-乙酰氨基己酸盐、烟酰胺和黄氧酸；涉及苯丙氨酸代谢、味觉传导、花生四烯酸代谢等20条代谢通路。结论 逍遥补肾方可以从多靶点、多成分、多代谢物、多代谢通路对HT产生调节作用。

基于网络药理学及代谢组学的逍遥补肾方治疗桥本甲状腺炎的作用机制

刘婧¹, 魏东升¹, 安杨¹, 孙欣², 伊桐凝³, 张兰³

1. 辽宁中医药大学研究生学院, 沈阳 110847;
2. 沈阳市第二中医医院内分泌科, 沈阳 110101;
3. 辽宁中医药大学附属医院内分泌科, 沈阳 110032

收稿日期:2023-04-28出版日期:2023-09-30发布日期:2023-09-02
通讯作者:张兰E-mail:zhanglanzhu@163.com
作者简介:刘婧(1992-),女,博士研究生.
基金资助:全国名老中医药专家传承工作室建设项目（国中医药人教函（2022）75号）


关键词: 逍遥补肾方, 桥本甲状腺炎, 网络药理学, 代谢组学
Abstract: Objective To explore the potential mechanism of Xiaoyao tonifying kidney formula in treating Hashimoto thyroiditis (HT). Methods The component targets of Xiaoyao tonifying kidney formula and the related disease targets of HT were intersected, and the protein-protein interaction (PPI) network and drug-component-intersection target networks were constructed using network pharmacology. Sixty-five rats were randomly divided into a control group (n=10) and an experimental autoimmune thyroiditis (EAT) model group (n=55). The EAT rat model was prepared by providing high-iodine water and multiple injections of thyroglobulin. After successful modeling, the EAT model rats were randomly divided into model, selenium yeast, and Xiaoyao tonifying kidney groups. The selenium yeast group was given 25.2 mg/(kg·d) selenium yeast tablet aqueous solution, the Xiaoyao tonifying kidney group was given 11.79 mg/(kg·d) Xiaoyao tonifying kidney formula solution, and the control and model groups were given equal volumes of distilled water. After continuous intragastric administration for 8 weeks, the serum of rats in each group was collected to detect the levels of ALB, AKT1, and TNF-α through enzyme-linked immunosorbent assay (ELISA). The feces of rats in each group were collected and tested for metabolomics using the ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method. Results A total of six core components of Xiaoyao tonifying kidney formula in treating HT were collected, including quercetin, kaempferol, β-sitosterol, stigmasterol, isorhamnetin, and luteolin. Five core targets containing ALB, AKT1, TNF, IL6, and TP53 were also collected. The results of these experiments showed that, compared with the selenium yeast group, the Xiaoyao tonifying kidney group could significantly downregulate the expression of ALB, AKT1, and TNF-α (P < 0.01). Metabolomic analysis identified five common differential metabolites, including (4Z, 7Z, 10Z, 13Z, 16Z, 19Z)-docosahexaenoic acid ethyl ester, (S)-2-methylmalate, 2-keto-6-acetamidocaproate, nicotianamine, and xanthoxic acid. Twentydifferent metabolic pathways were involved, such as phenylalanine metabolism, taste transduction, and arachidonic acid metabolism. Conclusion Xiaoyao tonifying kidney formula can regulate HT through multiple targets, components, metabolites, and metabolic pathways.
Key words: Xiaoyao tonifying kidney formula, Hashimoto thyroiditis, network pharmacology, metabolomics
PDF全文下载地址:

https://journal.cmu.edu.cn/CN/article/downloadArticleFile.do?attachType=PDF&id=3277