海藻、马尾藻广泛用于治疗甲状腺肿。甘草，即甘草根及其根茎，在传统中药中以减毒或增效而闻名。海藻甘草（HG）作为经典药对，是治疗甲状腺肿的常用且有效的联合疗法。 HG对甲状腺肿的潜在生物学机制尚不清楚。为探讨HG对甲状腺肿的影响，采用分子对接结合实验验证来阐明其分子机制。通过灌胃丙硫氧嘧啶（PTU）一段时间建立大鼠甲状腺肿模型14 天。将大鼠分为六组：对照组、模型组、优甲乐组、HG高剂量组（HG-H）、中剂量组（HG-M）和低剂量组（HG-L）。根据一般观察（如大鼠的生活状态、体重、直肠温度）、甲状腺相对重量、甲状腺功能、苏木精-伊红（HE）染色、透射电镜（TEM）观察病理变化。甲状腺的变化，评估了 HG 的效果。通过UPLC-MS/MS发现了HG的化学成分，并通过多个数据库预测了可能的目标。接下来，我们利用分子对接探索了它们的药理机制，并使用蛋白质印迹 (WB) 和免疫共沉淀 (Co-IP) 验证了关键靶点。HG 显着提高了三碘甲状腺原氨酸 (T3)、游离三碘甲状腺原氨酸 (FT3)、游离甲状腺素 (FT3) 的水平。 FT4)，PTU 诱导的大鼠体重增加并减少肿胀的甲状腺。模型组出现滤泡上皮细胞大小不均匀、形状不规则、排列紊乱等病理变化。然而，HG组的甲状腺滤泡和上皮细胞显得明显正常。与模型组相比，HG组中出现了多种自噬囊泡特征性变化。综上所述，HG-L显示出最佳的治疗效果。通过UPLC-MS/MS鉴定了HG的主要化学成分。结果表明，HG含有黄酮类、生物碱、有机酸、酚酸和萜类化合物等成分。芒柄花素和柚皮素与Beclin1蛋白的分子对接结果显示出-5.38 kcal/mol和-5.25 kcal/mol的高度相互作用。这意味着芒柄花素和柚皮素可能通过控制 Beclin1 介导的自噬对甲状腺肿大鼠产生治疗作用。 Western Blot (WB) 和免疫共沉淀 (Co-IP) 结果显示 HG 可以破坏 Beclin1/III 类磷脂酰肌醇 3-激酶 (PI3KC3) 结合并促进 Beclin1/B 细胞白血病/淋巴瘤-2 (Bcl-2) 复合物形成。综上所述，结果表明，通过减少 Beclin1/PI3KC3 形成和增加 Beclin1/Bcl-2 结合活性来抑制自噬。HG 通过调节 Beclin1 介导的自噬来改善丙硫氧嘧啶诱导的甲状腺肿，从而促进甲状腺细胞的自噬。版权所有 © 2024。由 Elsevier B.V. 出版

Haizao, Sargassum, is widely used to treat goiter. Gancao, Glycyrrhizae radix et rhizome, is renowned for reducing toxicity or increasing effects in traditional Chinese medicine. As a classic herb pair, Haizao-Gancao (HG) is a commonly used and effective combined therapy for goiter. The underlying biological mechanisms of HG on goiter is still unclear.To explore the effect of HG on goiter, employing molecular docking combined with experimental validation to elucidate the molecular mechanism.The rat goiter model was created by gavageing propylthiouracil (PTU) intragastrically for a duration of 14 days. The rats had been separated into six groups: control, model, euthyrox, HG high-dose (HG-H), medium-dose (HG-M) and low-dose (HG-L) group. Based on general observations (such as the rats' living status, body weight, and rectal temperature), the relative weight of the thyroid, thyroid function, hematoxylin-eosin (HE) staining, and transmission electron microscopy (TEM) to view the pathological variations of the thyroid glands, the effect of HG was evaluated. Discovered the chemical composition of HG by UPLC-MS/MS and the possible targets were predicted adopting several databases. Next, we explored their pharmacological mechanisms using molecular docking and validated key targets using western blotting (WB) and co-immunoprecipitation (Co-IP).HG significantly increased the levels of triiodothyronine(T3), free triiodothyronine (FT3), free thyroxine (FT4), gained body weight and reduced tumescent thyroid glands in PTU-induced rats. The model group pathological changes such as uneven size, irregular shape and disordered arrangement of follicular epithelial cells occurred. However, HG groups thyroid follicles and epithelial cells appeared apparently normal. A variety of characteristic changes of autophagic vesicles appeared in the HG groups as opposed to the model group. In conclusion, the HG-L showed the best therapeutic effect. By UPLC-MS/MS, the major chemical components of HG were identified. The result revealed that HG contained flavonoids, alkaloids, organic acids, phenolic acidsand and terpenoids, etc. The molecular docking results of formononetin and naringenin and Beclin1 protein showed a high interaction of -5.38 kcal/mol and -5.25 kcal/mol. This implies that formononetin and naringenin may have a therapeutic effect in goiter rats by controlling the Beclin1-mediated autophagy. Western Blot (WB) and co-immunoprecipitation (Co-IP) results showed that HG can disrupt Beclin1/class III phosphatidylinositol 3-kinase(PI3KC3) binding and promote Beclin1/B-cell leukemia/lymphoma-2(Bcl-2) complex formation. Taken together, results demonstrate that autophagy inhibition via reducing Beclin1/PI3KC3 formation and increasing Beclin1/Bcl-2 binding activity.HG ameliorates propylthiouracil-induced goiter by regulating the Beclin1-mediated autophagy, thus promoting the autophagy of thyroid cells.Copyright © 2024. Published by Elsevier B.V.