H_(2)S对Graves甲亢小鼠甲状腺功能与甲状腺滤泡上皮细胞损伤的保护作用及机制  

Protective effect and mechanism of H2S on thyroid function and thyroid follicular epithelial cell damage in Graves hyperthyroid mice

作　　者：欧慧婷[1] 汪新宇[1] 余怀新[1] 陈泽龙[1] 李明政[1] OU Huiting;WANG Xinyu;YU Huaixin;CHEN Zelong;LI Mingzheng(Department of Endocrinology, Shenzhen Second People's Hospital, Shenzhen 518035, Guangdong, China)

机构地区：[1]深圳市第二人民医院内分泌科,广州深圳518035

出　　处：《西部医学》2021年第8期1138-1144,共7页Medical Journal of West China

基　　金：深圳市科技计划项目(JCYJ20150330102720141)。

摘　　要：目的探讨气体信号分子硫化氢(H_(2)S)对Graves甲亢(GD)小鼠甲状腺功能的改善情况,及其对甲状腺滤泡上皮细胞损伤的保护作用。方法45只BALB/c小鼠随机分为正常对照组、空载体病毒组和重组腺病毒组,每组15只。通过注射Ad-TSHR289重组腺病毒建立GD小鼠模型,GD造模成功的15只小鼠随机分为模型组、H_(2)S组及甲疏咪唑组,每组5只。H_(2)S组小鼠予以硫氢化钠(NaHS)溶液(56μmol/kg/d)腹腔注射,甲疏咪唑组小鼠予以甲疏咪唑溶液(3.9 mg/kg/d)腹腔注射,正常对照组和空载体病毒组小鼠予以等体积0.9%Nacl溶液腹腔注射,共干预4周。4周后,测定小鼠血清中促甲状腺激素受体抗体(TRAb)、甲状腺素(T4)和促甲状腺激素(TSH)水平,HE染色观察各组小鼠甲状腺组织的病理变化,免疫组织化学染色检测甲状腺组织增殖细胞核抗原(PCNA)与细胞周期蛋白D1(CyclinD1)表达,Western blot检测钠/碘转运体(NIS)、甲状腺过氧化物酶(TPO)、甲状腺球蛋白(Tg)表达水平。分离培养小鼠甲状腺滤泡上皮细胞,将细胞随机分为正常对照组、H_(2)O_(2)组、NaHS+H_(2)O_(2)组,正常对照组细胞不进行任何处理,H_(2)O_(2)组使用H_(2)O_(2)溶液处理甲状腺滤泡上皮细胞24 h,NaHS+H_(2)O_(2)组先用NaHS预处理甲状腺滤泡上皮细胞30 min再用H_(2)O_(2)溶液处理细胞。通过CCK-8法检测各组细胞活性,流式细胞术检测各组细胞凋亡情况。结果与正常对照组比较,重组腺病毒组小鼠血清中TRAb和T4水平显著上升,TSH水平显著下降(均P<0.05)。NaHS溶液干预后,GD小鼠血清中TRAb和T4的水平显著下降,TSH水平显著升高(均P<0.05);甲状腺组织病变得到明显的改善,PCNA与CyclinD1阳性表达显著降低,NIS、TPO与Tg蛋白表达量显著下降(均P<0.05)。与正常对照组比较,H_(2)O_(2)组甲状腺滤泡上皮细胞活性在培养24、48、72 h显著下降,细胞凋亡率显著升高(均P<0.05);与H_(2)O_(2)组比较,NaHS+H_(2)O_(2)组细�Objective To explore the improvement of thyroid function of gas signal molecule hydrogen sulfide(H_(2)S)in Graves hyperthyroidism(GD)mice and its protective effect on thyroid follicular cell damage.Methods 45 BALB/c mice were randomly divided into normal control group,empty vector virus group and recombinant adenovirus group.GD mouse model was established by injection of Ad-TSHR289 recombinant adenovirus.The mice successfully modeled by GD were randomly divided into model group,H_(2)S group and methabazole group.The mice in the imidazole group were given intraperitoneal injection of mesozole solution(3.9mg/kg/d),and the mice in the normal control group and the empty vector virus group were given intraperitoneal injection of an equal volume of 0.9%Nacl solution for 4 weeks.After 4 weeks,serum thyrotropin receptor antibody(TRAb),thyroxine(T4)and thyroid stimulating hormone(TSH)levels were measured.HE staining was used to observe the pathological changes of thyroid tissue in each group of mice.Immunohistochemical staining was used to detect the expression of proliferating cell nuclear antigen(PCNA)and cyclin D1(CyclinD1)in thyroid tissue.Western blot was used to detect the expression levels of sodium/iodine transporter(NIS),thyroid peroxidase(TPO)and thyroglobulin(Tg).Isolate and culture mouse thyroid follicular epithelial cells,and randomly divide the cells into normal control group,H_(2)O_(2) group and NaHS+H_(2)O_(2) group.The cells in the normal control group did not undergo any treatment.The H_(2)O_(2) group used H_(2)O_(2) solution to treat the thyroid follicular epithelial cells for 24 hours.The NaHS+H_(2)O_(2) group first treated the thyroid follicular epithelial cells with NaHS for 30 minutes and then treated the cells with H_(2)O_(2) solution.The cell activity of each group was detected by CCK-8 method,flow cytometry was used to detect the apoptosis of each group.Results Compared with the normal control group,the levels of TRAb and T4 in the serum of mice in the recombinant adenovirus group increased signi

关 键 词：硫化氢 GRAVES甲亢 甲状腺组织 甲状腺滤泡上皮细胞 

分 类 号：R581.1[医药卫生—内分泌]