Effect of Huatan Sanjie Formula(化痰散结方)on Thyroid Angiogenesis and VEGFA/VEGFR2 Signaling Path-way in Graves'Disease Model Mice
Objective To investigate the possible mechanism of Huatan Sanjie Formula(化痰散结方,HSF)in treating Graves'disease(GD)from the perspective of thyroid angiogenesis.Methods Thirty-six BALB/c female mice were randomly divided into a normal control group(n=9)and a modeling group(n=27).Mice in the modeling group were injected with 2.0×109 PFU/ml of Ad-TSHR289 adenovirus into the tibialis anterior muscle to build GD model.Nine weeks after immunization,the successfully modeled mice were randomly divided into model group,methimazole(MMI)group and HSF group,with 9 mice in each group.The MMI group was given 5.2 mg/(kg·d)of methimazole tablets by gavage,while the HSF group was given HSF at a relative crude drug dosage of 7.02 g/(kg·d)by gavage.The normal control group and the model group were given 0.1 ml/10 g of pure water by gavage.All groups were administered intragastrically once a day for a total of 4 weeks.The levels of thyroxine(T4)and thyrotropin receptor autoantibodies(TRAb)in serum were detected by radioimmunoassay,while the pathological changes of the thyroid gland were assessed by HE staining.The vascular morphology of thyroid tissue was observed by CD34 immuno-histochemical staining,and the microvessel density(MVD)was counted.The protein expression of vascular endothe-lial growth factor A(VEGFA)and vascular endothelial growth factor receptor 2(VEGFR2)in thyroid was detected by Western-blot.Results Compared to those in the normal control group,the thyroid volume of the mice in the model group significantly increased with excessive congestion,and the pathology showed significant thyroid follicular hyper-plasia,columnar and proliferated epithelial cells,and enlarged follicle size;serum T4 and TRAb significantly in-creased,as well as the count of thyroid MVD,and the protein expressions of thyroid VEGFA and VEGFR2(P＜0.01).Compared to those in the model group,the thyroid glands of the mice in the MMI group and the HSF group were significantly reduced,and the congestion was improved;pathology showed that thyroid follicular hyperplasia and epithelial cell proliferation were reduced,with smooth edges of the follicles and the significantly reduced inward pro-trusion;serum T4 and TRAb significantly decreased,as well as the thyroid MVD,thyroid VEGFA and VEGFR2 pro-tein expressions(P＜0.05 or P＜0.01).There was no significant difference in all indicators between the MMI group and the HSF group(P＞0.05).Conclusion HSF may inhibit thyroid angiogenesis by down-regulating thyroid VEGFA/VEGFR2 signaling pathway,thereby improving goitre and hyperfunction in GD mice.
万方数据
维普
