四逆散调控Hippo信号通路改善Graves病甲状腺肿大的机制研究OA

Objective To investigate the effects of Sinisan on thyroid goiter in Graves'disease(GD)and explore its underlying mechanism involving the regulation of the Hippo signaling pathway and proliferative factors.Methods Forty-eight female BALB/c mice were randomly divided into six groups(n=8).GD models were established in five groups by intramuscular injection of Ad-TSHR289 adenovirus,with the non-injected group serving as the normal control.After successful model establishment,the modeled mice were randomly divided into the following groups:model group,methimazole group(4.55 mg/kg),and low-,medium-,and high-dose Sinisan groups(2.73,5.46,10.92 g/kg).Interventions were administered for 6 weeks.General thyroid morphology was observed;the thyroid coefficient was calculated;pathological changes in thyroid follicular epithelial cells were assessed by hematoxylin-eosin(HE)staining;serum levels of thyrotropin receptor antibody(TRAb)and thyroxine(T4)were measured by ELISA;The mRNA and protein expression levels of proliferation markers(Ki-67 and PCNA)and key factors of the Hippo signaling pathway(pMST1,MST1,YAP,and TAZ)were detected using quantitative real-time PCR and Western blot analysis.Results Compared with the normal group,the model group exhibited significantly enlarged thyroids,hyperplastic thyroid follicular epithelial cells with irregular morphology,and markedly increased thyroid coefficient and serum TRAb and T4 levels(all P<0.01).Compared with the model group,the methimazole group and all Sinisan dose groups significantly alleviated thyroid goiter,improved follicular epithelial cell hyperplasia,and reduced thyroid coefficient(P<0.01,P<0.05)and serum TRAb and T4 levels(all P<0.01).Among these groups,the medium-dose Sinisan group showing the most pronounced improvement in histology of the thyroid gland,thyroid coefficient and TRAb levels.Consequently,this group was selected for subsequent mechanistic studies.Compared with the normal group,the model group showed significantly upregulated mRNA expression of proliferation-related factors PCNA and Ki-67,as well as upregulated PCNA protein expression(P<0.01,P<0.05).Compared with the model group,the medium-dose Sinisan group significantly downregulated the mRNA expression of PCNA and Ki-67(P<0.01)and PCNA protein expression(P<0.05).Compared with the normal group,the model group exhibited significantly downregulated mRNA expression of the key Hippo pathway factor MST1(P<0.05),significantly upregulated mRNA expression of YAP and TAZ(P<0.01,P<0.05),significantly upregulated protein expression of YAP and TAZ(P<0.01),and significantly downregulated expression of MST1 and pMST1(P<0.01,P<0.05).Compared with the model group,the medium-dose Sinisan group significantly downregulated YAP and TAZ mRNA and protein expression(P<0.01,P<0.05,P<0.05),upregulated MST1 mRNA and protein expression(P<0.05,P<0.01),and showed an upward trend in pMST1 protein expression,although this difference was not statistically significant(P>0.05).Conclusion Sinisan effectively ameliorates thyroid goiter in GD.Its mechanism is associated with the inhibition of thyroid cell proliferation through the regulation of the Hippo signaling pathway.