Haizao-gancao草药对通过调节beclin1介导的自噬来改善丙酸硫酸甲酸酯诱导的甲状腺肿

Sargassum Haizao广泛用于治疗甲状腺肿。 Gancao，Glycyrrhizae Radix et Rhizome，以降低传统中药的毒性或增加影响而闻名。作为经典的草药对，Haizao-Gancao（HG）是一种常用且有效的联合疗法。 HG在甲状腺肿上的基本生物学机制仍然不清楚。要探索Hg对甲状腺肿的影响，采用分子对接结合了实验验证来阐明分子机制。大鼠甲状腺癌模型是通过内部内部的propylthiouracil（ptu）在内部内部进行14天的持续时间创建的。将大鼠分为六组：对照，模型，Euthyrox，Hg高剂量（HG-H），中剂量（HG-M）和低剂量（HG-L）组。基于一般观察（例如大鼠的生活状况，体重和直肠温度），甲状腺功能，甲状腺素蛋白蛋白（HE）染色和透射电子显微镜（TEM）的相对重量，以查看甲状腺的病理变化，评估了HG的效应。通过UPLC-MS/MS发现了HG的化学成分，并且预计可能采用了几个数据库。接下来，我们使用分子对接和使用蛋白质印迹（WB）和共免疫沉淀（CO-IP）探索了他们的药理机制。 PTU引起的大鼠。模型组的病理变化，例如不均匀的大小，不规则形状和卵泡上皮细胞的无序排列。然而，HG组甲状腺卵泡和上皮细胞显然显然是正常的。自噬囊泡的各种特征变化出现在HG组中，而不是模型组。总之，HG-L显示出最佳的治疗作用。通过UPLC-MS/MS，确定了HG的主要化学成分。结果表明，HG含有类黄酮，生物碱，有机酸，酚类酸和萜类化合物等。Formononetin和naringenin和beclin1蛋白的分子对接结果表现出-5.38 kcal/mol和-5.25 kcal/mol的高相互作用。这意味着formononetin和naringenin可能通过控制beclin1介导的自噬来对甲状腺肿大鼠具有治疗作用。 Western印迹（WB）和共免疫沉淀（CO-IP）结果表明，HG可以破坏Beclin1/III III磷脂酰肌醇3-激酶（PI3KC3）结合，并促进Beclin1/B-细胞白血病/lymphoma/Lymphoma/Lymphoma/Lymphoma-2（Bcl-2）复合物。综上所述，结果表明，通过减少beclin1/pi3kc3的形成并增加beclin1/bcl-2结合活性来自噬抑制作用。Hg通过调节beclin1介导的自噬来缓解丙曲霉诱导的甲状腺癌，从而促进甲状腺细胞的自噬菌。

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.