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目的:本实验以Fischer大鼠甲状腺细胞系(FRTL)为研究对象,在体外从细胞和分子水平研究高糖毒性及AGEs对甲状腺细胞甲状腺球蛋白(TG)、甲状腺过氧化物酶(TPO)mRNA表达的影响,初步探索除免疫机制外,糖毒性致甲状腺组织损伤的机制。方法:在体外用D-glucose与BSA共同孵育AGEs-BSA。将体外培养成功的FRTL细胞分为+TSH组(完全培养基中加TSH)和-TSH组(完全培养基中未加TSH),每组分别给予不同浓度的D-glucose、AGEs及AGEs抗体刺激,在糖刺激组设置渗透组以排除渗透压因素对细胞的影响,于24h、48h、72h分别收集细胞,使用实时荧光定量PCR测定TG、TPO mRNA的表达情况。结果:①12周后得到棕褐色产物AGEs-BSA。AGEs-BSA荧光光谱扫描:其激发波长为360nm,发射波长为450nm,AGEs-BSA的荧光值高于BSA,其浓度为52.446g/ml。②实验中FRTL细胞无污染,圆形较亮,成堆生长,密度适中,贴壁良好。③实时荧光定量PCR检测TG、TPOmRNA的表达水平:在+TSH组与-TSH组:D-glucose刺激组及AGEs刺激组中,每个刺激时间中组间比较均有统计学意义(P＜0.001);且在D-glucose刺激组,对照组与渗透组比较差别无统计学意义(P＞0.05);+TSH组不同浓度D-glucose刺激24h、48h、72h后;15mmol/L组低于对照组、渗透组(TG、TPO-24h:P＞0.05:48h:P＜0.001;72h:P＜0.001);20mmol/L组低于15mmol/L组、对照组、渗透组(TG、TPO:24h:P＞0.05、P＜0.001;48h:P＜0.001、P=0.908,0.000,0.000;72h:P=0.910,0.000,0.000、P=0.513,0.000,0.000);30mmol/L组明显低于15mmol/L组、20mmol/L组、对照组、渗透组(TG、TPO:24h:P＜0.001;48h:P=0.289,0.314,0.000,0.000、P＜0.001;72h:P=0.723,0.818,0.000,0.000、P=0.000,0.000,0.484,0.962);-TSH组不同浓度D-glucose刺激细胞24h、48h、72h后:15mmol/L组低于TSH刺激组、对照组、渗透组(TG,TPO:24h、48h、72h:P＜0.001);20mmol/L组低于15mmol/L组、TSH刺激组、对照组、渗透组(TG、TPO:24h:P=0.022,0.000,0.000,0.000、P=0.871,0.000,0000,0000;48h:P=0.855,0.000,0.000,0.000、P=0.383,0.000,0.000,0.000;72h:P=0.657,0.000,0.000,0.000、P=0.878,0.000,0.000,0.000);30mmol/L组明显低于15mmol/L组、20mmol/L组、TSH刺激组、对照组、渗透组(TG、TPO:24h:P=0.001,0.100,0.000,0.000,0.000、P＜0.05;48h:P=0.859,0.9960.000,0.000,0.000、P=0.131,0.027,0.000,0.000,0.000;72h:P=0.002,0.005,0.000,0.000,0.000、P=0.059,0.045,0.000,0.000,0.000);+TSH组不同浓度AGEs分别刺激细胞24h、48h、72h后:0.25mg/dl组低于AGEs抗体组、对照组(TG、TPO:24h:P＜0.001、P＜0.001;48h:P＜0.001、P＜0.001;72h:P=0.685,0.000、P＜0.001);0.5mg/dl组低于0.25mg/dl组、对照组(TG、TPO:24h:P=0.254,0.000、P＜0.001;48h:P=0.669,0.000、P=0.229,0.000;72h:P=0.658,0.000、P=0.539,0.000);1.0mg/dl组低于0.25mg/dl组、0.5mg/dl组、对照组(TG、TPO:24h:P=0.435,0.700,0.000、P=0.000,0.993,0.000;48h:P=0.002,0.003,0.003、P＜0.001;72h:P=0.364,0.631,0.000、P＜0.001);-TSH组不同浓度AGEs刺激细胞24h、48h、72h后:0.25mg/dl组低于AGEs抗体组、TSH刺激组、对照组(TG、TPO:24h、48h、72h:P＜0.001);0.5mg/dl组低于0.25mg/dl组、TSH刺激组、对照组(TG、TPO:24h:P=0.945,0.000、0.000、P＜0.001;48h:P＜0.001、P=0.001,0.000,0.000;72h:P=0.973,0.000,0.000、P=0.970,0.000,0.000):1.0mg/dl组低于0.25mg/dl组、0.5mg/dl组、TSH刺激组、对照组(TG、TPO:24h:P=0.887,0.945,0.000,0.000、P=0.000,0.535,0.000,0.000;48h:P=0.887,0.594,0.000,0.000、P=0.001,0.893,0.000,0.000;72h:P＜0.001、P＜0.001)。结论:不同浓度的D-glnoose组及AGEs组的TG、TPOmRNA的表达较对照组有下降的趋势,且随着D-glucose、AGEs浓度的升高,二者的表达下降趋势更明显,从体外进一步证明高糖和AGEs对甲状腺细胞有一定的损害,AGEs与高糖可产生相似的作用,且提示高糖对甲状腺的损伤可能独立于免疫机制;在TSH刺激组,二者的表达明显高于对照组及其它各实验组,提示甲状腺功能变化受TSH的影响;在AGEs抗体组,二者的表达较单纯AGEs组有所升高,提示AGEs可能是高糖引起甲状腺损伤的重要因素之一。