摘要
目的研究桥本甲状腺炎(HT)对孕鼠额叶谷氨酸(Glu)浓度的影响及Glu转运体的变化。方法适应性饲养50只NOD雌性小鼠1周后,随机分为正常对照组(CON组)20只、桥本甲状腺炎组(HT组)30只,将猪甲状腺球蛋白(pTg)于HT组小鼠尾根部皮下注射制备HT小鼠模型。造模成功后,各组雌鼠与健康NOD雄鼠合笼使其受孕。通过ELISA测定血清促甲状腺激素(TSH)水平;电化学发光免疫分析法测定血清甲状腺过氧化物酶抗体(TPOAb)、甲状腺球蛋白抗体(TgAb)水平和血清甲状腺激素三碘甲状腺原氨酸(T3)、甲状腺素(T4)水平;ELISA法检测孕鼠额叶Glu浓度;Western blot法检测额叶中囊泡谷氨酸转运体(VGLUT)1和兴奋性谷氨酸转运体(EAAT)2的蛋白表达水平。RTqPCR观察孕鼠额叶VGLUT 1、EAAT 2 mRNA的表达水平。结果①HT组孕鼠血清甲状腺抗体水平(TPOAb、TgAb)较CON组升高(P<0.05),2组血清T3、T4、TSH差异无统计学意义;②ELISA法显示:与CON组比较,HT组额叶中Glu含量升高(P<0.05);③Western blot法:HT组VGLUT 1蛋白表达较CON组降低(P<0.05),2组EAAT 2的蛋白表达水平差异无统计学意义;④RT-qPCR法显示:HT组额叶VGLUT 1 mRNA表达相比CON组降低(P<0.05),2组EAAT 2 mRNA表达水平差异无统计学意义。结论HT可导致孕鼠额叶Glu含量增高,Glu转运体VGLUT 1蛋白表达下降,提示HT可导致Glu代谢紊乱。
Objective To investigate the effects of Hashimoto's thyroiditis(HT)on the concentration of glutamate(Glu)in the frontal lobe of pregnant rats and the changes of glutamate transporters.Methods After adaptively feeding for a week,50 NOD female micewere randomly divided into normal control group(CON group,20 mice)and Hashimoto thyroiditis group(HT group,30 mice).HT mouse model was prepared by subcutaneous injection of porcine thyroglobulin in the tail root of HT group.After successful modeling,female mice in each group were caged with healthy NOD males to get pregnant.Serum thyroid stimulating hormone(TSH)levels were determined by ELISA.The levels of serum thyroid peroxidase antibody(TPOAb),thyroglobulin antibody(TgAb)and serum thyroid hormone triiodothyronine(T3)and thyroxine(T4)were determined by electrochemiluminescence immunoassay.The concentration of Glu in the frontal lobe of pregnant rats was detected by ELISA.Western blot assay was used to detect protein expression levels of vesicular glutamate transporter VGLUT 1 and excitatory glutamate transporter EAAT 2 in frontal lobe.RT-qPCR was used to observe the expression of VGLUT1 and EAAT2 mRNA in the frontal lobe of pregnant rats.Results①Serum thyroid antibody levels(TPOAb,TgAb)in HT group were higher than those in CON group(P<0.05).There was no significant difference in serum T3,T4 and TSH between the two groups;②The ELISA showed that the glutamate content in the frontal lobe of the HT group was higher than that in the CON group(P<0.05);③Western blot method showed that the expression of VGLUT 1 protein in HT group was lower than that in CON group(P<0.05),and the protein expression level of EAAT 2 in the two groups was not statistically significant;④RT-qPCR showed that the mRNA expression of VGLUT 1 in the frontal lobe of HT group was lower than that in CON group(P<0.05).There was no significant difference in the expression level of EAAT 2 mRNA between the two groups.Conclusion HT can increase the content of Glu in the frontal lobe of pregnant mice,and decrease the expression of the glutamate transporter VGLUT 1 protein,suggesting that HT can cause glutamate metabolism disorders.
作者
柳田田
杨昊
夏琴
程锦
吴章碧
王囡
朱德发
Liu Tiantian;Yang Hao;Xia Qin(Dept of Elderly Endocrinology,The First Affiliated Hospital of Anhui Medical University,Hefei 230022)
出处
《安徽医科大学学报》
CAS
北大核心
2020年第5期693-697,共5页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金(编号:81272152)。
关键词
桥本甲状腺炎
额叶
谷氨酸
谷氨酸转运体
怀孕
Hashimoto's thyroiditis
frontal lobe
glutamic acid
glutamate transporter
pregnancy
