Xiao-Luo-Wan treats propylthiouracil-induced goiter with hypothyroidism in rats through the PI3K-AKT/RAS pathways based on UPLC/MS and network pharmacology
Xiao-Luo-Wan treats propylthiouracil-induced goiter with hypothyroidism in rats through the PI3K-AKT/RAS pathways based on UPLC/MS and network pharmacology
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DOI：
10.1016/j.jep.2022.115045
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发表时间：
2022-02-03
影响因子：
5.4
通讯作者：
Shi, Yuanyuan
中科院分区：
文献类型：
--
作者：
Dong, Shi;Liu, Qunying;Shi, Yuanyuan
民族药理学相关性：甲状腺肿合并甲状腺功能减退症发生在多种甲状腺疾病中。小螺丸（XLW），含有玄参、贝母。牡蛎（Ostrea gigas Thunberg）在中国作为治疗甲状腺肿的有效中药已有数百年历史。根据临床观察和实验研究，XLW对甲状腺功能减退症也有一定的疗效。然而，XLW的治疗机制仍不清楚。研究目的：本研究旨在探讨XLW对丙硫氧嘧啶（PTU）诱发的甲状腺肿伴甲状腺功能减退大鼠的治疗作用，并利用超高效液相色谱-质谱（UPLC/MS）、网络药理学和分子对接模拟揭示其潜在的分子机制。材料与方法：造模成功后，将其余大鼠随机分为模型组、优甲乐组、XLW组和对照组。连续灌胃相应药物4周。监测生长状况，计算甲状腺相对重量，检测血清总T3、T4、TSH含量。苏木精-伊红(H&E)染色观察甲状腺病理变化。通过 UPLC/MS 鉴定了 XLW 的化学成分，并使用多个数据库预测了 XLW 的假定目标。我们基于甲状腺肿/甲状腺功能减退症相关目标和 XLW 目标的交叉点进行了网络药理学。然后，我们进行了KEGG通路富集分析，并利用蛋白质-蛋白质相互作用（PPI）网络进一步筛选关键靶标。最后，利用分子对接来预测XLW识别的成分与关键靶标的结合能力。结果：XLW 显着提高 PTU 诱导大鼠的 T3 和 T4 水平，降低 TSH，增加体重，并减少肿胀的甲状腺。 XLW促进甲状腺滤泡和上皮细胞的形态恢复。使用 UPLC/MS 鉴定了 XLW 的 21 种主要化学成分。通过检索传统中药系统药理学数据库（TCMSP）、Swiss Target Prediction和UniProt数据库，获得了270个XLW的潜在基因靶点和717个甲状腺肿/甲状腺功能减退症的已知靶点。总共 83 条 KEGG 通路富含磷脂酰肌醇 3-激酶-蛋白激酶 B (PI3K-AKT) 和 RAS 信号通路。 PPI 分析揭示了激酶蛋白激酶 B (AKT) 1、白介素 (IL) 6、血管内皮生长因子 A (VEGFA)、肿瘤坏死因子 (TNF)、甘油醛 3-磷酸脱氢酶 (GAPDH)、表皮生长因子受体 (EGFR)、GTPase HRas (HRAS)、基质金属蛋白酶 (MMP) 9 和热休克蛋白 90 α 家族 A 类成员 1 的九个关键靶点(HSP90AA1)。分子对接验证了哪些药物成分对关键靶点具有良好的结合能力（全部
Ethnopharmacological relevance: Goiter with hypothyroidism occurs in several thyroid diseases. Xiao-Luo-Wan (XLW), which contains Scrophularia ningpoensis Hemsl., Fritillaria thunbergii Miq. and Ostrea gigas Thunberg, has been used as an effective Chinese medicine for the treatment of goiters in China for hundreds of years. Based on clinical observations and experimental studies, XLW also exerts a certain effect on hypothyroidism. However, the therapeutic mechanism of XLW remains unclear. Aim of the study: The present study aimed to investigate the therapeutic effect of XLW on propylthiouracil (PTU)induced goiter with hypothyroidism in rats and to uncover the underlying molecular mechanism using ultra high-performance liquid chromatography-mass spectrometry (UPLC/MS), network pharmacology, and molecular docking simulations. Materials and methods: After successful modeling, the remaining rats were randomly divided into a model group, an Euthyrox group, an XLW group, and a control group. The corresponding drugs were given by gavage for four consecutive weeks. The growth status was monitored, the relative thyroid weight was calculated, and the total serum T3, T4, and TSH content were detected. Hematoxylin-eosin (H & E) staining was used to observe the pathological changes in the thyroid glands. The chemical components of the XLW were identified by UPLC/MS and the putative targets of XLW were predicted using multiple databases. We performed network pharmacology based on the intersection of goiter/hypothyroidism-related targets and XLW targets. Then, we performed KEGG pathway enrichment analysis, and key targets were further screened using protein-protein interaction (PPI) networks. Finally, molecular docking was used to predict the binding ability of XLW identified components and the key targets. Results: XLW significantly increased the levels of T3 and T4, and reduced TSH, increased body weight, and decreased swollen thyroid glands in PTU-induced rats. XLW promoted the morphological recovery of thyroid follicles and epithelial cells. Twenty-one main chemical components of XLW were identified using UPLC/MS. 270 potential gene targets of XLW and 717 known targets of goiter/hypothyroidism disease were obtained by searching the Traditional Chinese Medicine Systems Pharmacology Database (TCMSP), Swiss Target Prediction, and UniProt databases. A total of 83 KEGG pathways were enriched with phosphatidylinositol 3-kinase-protein kinase B (PI3K-AKT) and RAS signaling pathways. PPI analysis revealed nine key targets of kinase-protein kinase B (AKT) 1, interleukin (IL) 6, vascular endothelial growth factor A (VEGFA), tumor necrosis factor (TNF), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), epidermal growth factor receptor (EGFR), GTPase HRas (HRAS), matrix metalloproteinase (MMP) 9, and heat shock protein 90 alpha family class A member 1 (HSP90AA1). Molecular docking verified which drug components had good binding ability to key targets (all