化痰散结方对Graves病模型小鼠甲状腺血管新生及VEGFA/VEGFR2信号通路的影响  

Effect of Huatan Sanjie Formula(化痰散结方)on Thyroid Angiogenesis and VEGFA/VEGFR2 Signaling Pathway in Graves'Disease Model Mice

作　　者：马文欣 朱晓云[2] 王程娜 徐婧 刘喜明[2] 汤阳 MA Wenxin;ZHU Xiaoyun;WANG Chengna;XU Jing;LIU Ximing;TANG Yang(Beijing University of Chinese Medicine,Beijing,100029;Guang,anmen Hospital,China Academy of Chinese Medical Sciences;Dongzhimen Hospital,Beijing University of Chinese Medicine)

机构地区：[1]北京中医药大学,北京市100029 [2]中国中医科学院广安门医院 [3]北京中医药大学东直门医院

出　　处：《中医杂志》2024年第19期2025-2031,共7页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(82305182,81774293);中国博士后科学基金资助项目(2023M740347)。

摘　　要：目的 从甲状腺血管新生角度探讨化痰散结方治疗Graves病的可能作用机制。方法 36只BALB/c雌性小鼠随机分为正常对照组9只和造模组27只,造模组小鼠以Ad-TSHR289腺病毒2.0×10^(9) PFU/ml胫前肌肉注射100μl构建Graves病小鼠模型。免疫后第9周将造模成功小鼠随机分为模型组、甲巯咪唑组和化痰散结组,每组9只。甲巯咪唑组小鼠给予甲巯咪唑片5.2 mg/(kg·d)灌胃,化痰散结组小鼠给予化痰散结方相对生药量7.02 g/(kg·d)灌胃,正常对照组与模型组给予纯水0.1 ml/10 g灌胃。均每日灌胃1次,共计4周。放射免疫法检测各组小鼠血清中甲状腺素(T4)和促甲状腺素受体自身抗体(TRAb)含量,HE染色观察甲状腺病理改变情况,CD34免疫组化染色观察甲状腺组织血管形态并对微血管密度(MVD)计数,Westernblot法检测小鼠甲状腺中血管内皮生长因子A (VEGFA)和血管内皮生长因子受体2 (VEGFR2)蛋白表达。结果 与正常对照组比较,模型组小鼠甲状腺体积明显增大,存在过度充血,病理显示甲状腺滤泡增生显著,上皮细胞呈柱状且增殖明显,滤泡体积增大;血清中T4、TRAb水平上升,甲状腺MVD计数增多,甲状腺VEGFA、VEGFR2蛋白表达上调(P<0.01)。与模型组比较,甲巯咪唑组和化痰散结组小鼠的甲状腺明显减小,充血情况有所改善,病理显示甲状腺滤泡增生和上皮细胞增殖减轻,滤泡边缘光滑、内凸明显减少;血清T4、TRAb水平降低,甲状腺MVD计数减少,甲状腺VEGFA、VEGFR2蛋白表达量显著下调(P<0.05或P<0.01)。甲巯咪唑组和化痰散结组各指标差异均无统计学意义(P>0.05)。结论 化痰散结方可能通过下调甲状腺VEGFA/VEGFR2信号通路从而抑制甲状腺血管新生,改善Graves病甲状腺肿大和功能亢进。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 2.0×10^(9)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 immunohistochemical staining,and the microvessel density(MVD)was counted.The protein expression of vascular endothelial 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 hyperplasia,columnar and proliferated epithelial cells,and enlarged follicle size;serum T4 and TRAb significantly increased,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

关 键 词：GRAVES病 甲状腺 血管新生 血管内皮生长因子 化痰散结方 

分 类 号：R285.5[医药卫生—中药学]