Mechanism of Huatan Sanjie Fang in improving goiter in Graves' disease mice based on the Hippo signaling pathway

Abstract

To explore the mechanism of Huatan Sanjie Fang (HTSJ) in regulating goiter in Graves’ disease (GD) mice by detecting key factors of the hippo signaling pathway. A mouse model of GD was established by injecting Ad-TSHR289 adenovirus into the bilateral quadriceps femoris of female mice. Successful mouse models were then randomly divided into a model group, methimazole (MMI) group, and HTSJ group, and fed with deionized water, MMI (4.5 mg/kg per day), and HTSJ (35.10 g/kg per day), respectively, for 10 weeks. Histopathological changes of the thyroid gland were subsequently observed by hematoxylin-eosin staining. Radioimmunoassay was used to detect serum total thyroxine (T4) and thyrotrophin-receptor antibody (TRAb) levels. The relative expression of mRNA of Mst1, YAP, and TAZ was detected by quantitative real-time polymerase chain reaction, while the protein expression of Mst1, YAP, TAZ, pMst1, and pYAP was detected by western blot. After 10 weeks of drug intervention, goiter and other pathological changes in the HTSJ group significantly improved compared with the model group, and the levels of serum T4 and TRAb significantly decreased (P = .002, P < .001, respectively). Decreased mRNA expression of Mst1, YAP, and TAZ, the key factors of the hippo signaling transduction pathway, was also observed (P = .002, P = .022, P < .001, respectively). In contrast, protein expression of Mst1 (P = .046), pMst1 (P = .026), and pYAP (P = .004) increased, while protein expression of YAP and TAZ decreased (P = .041, P < .001, respectively). HTSJ can effectively improve goiter in GD mice through the hippo signaling pathway.