论文部分内容阅读
目的 研究甲状腺刺激性抗体 (TSAb)对甲状腺细胞合成和分泌激素功能的影响以及通过的信号传导途径 ,初步探讨 Graves’病 (GD)发病的分子机制。 方法 应用原代培养人甲状腺细胞为研究对象 ,放射免疫法检测 TSAb刺激后培养介质 T3、T4 、甲状腺球蛋白 (TG)的生成 ,改良愈创木酚氧化法检测细胞的甲状腺过氧化物酶 (TPO)活性 ;同时应用信号传导抑制剂阻断各传导途径 ,观察蛋白激酶 (PKA)抑制剂、细胞内 Ca2 +鳌合剂 (BAPTA- AM)、酪氨酸蛋白激酶(TPK)抑制剂——木黄酮对 T3、T4 、TG生成及 TPO活性的影响。 结果 TSAb阳性组呈浓度和时间依赖性刺激 T3、T4 、TG生成及 TPO活性增加 ,T3、T4 、TG分泌高峰分别为2 mg/ m L TSAb 作用 4 8h,2 mg/ m L TSAb 作用 4 8h,3mg/ m L TSAb作用 72 h;TPO活性的高峰为 4 mg/ m LTSAb作用 4 8h。同时加 PKA抑制剂阻断环磷酸腺苷 /蛋白激酶 A (c AMP/ PKA )途径后 ,T3生成降低 6 3.3% ,T4 降低6 7.9% ,TG降低 5 1 .5 % ,TPO活性减少 79.2 % ;加 BAP-TA - AM鳌合细胞内 Ca2 + 阻断 PIP2 / Ca2 + 途径 ,能部分抑制T3、T4 、TG的生成 ,但对 TPO活性无明显作用 (P >0 .0 5 ) ;TPK通道抑制剂木黄酮对 T3、T4 、TG、TPO活性均无明显影响 (P>0 .0 5 )。 结论 (1 ) TSAb可
Objective To investigate the effects of thyroid stimulating antibody (TSAb) on thyroid cell synthesis and secretion of hormones and the signaling pathways involved in the pathogenesis of Graves’ disease (GD). Methods Primary cultured human thyrocytes were used as research objects. Radioimmunoassay was used to detect the production of T3, T4 and thyroglobulin (TG) after TSAb stimulation. The activity of thyroid peroxidase TPO). At the same time, the signal transduction inhibitors were used to block the various pathways. The effects of PKA inhibitors, BAPTA-AM and TPK inhibitors Effects of Flavonoids on T3, T4, TG Production and TPO Activity. Results The TSAb-positive group stimulated the production of T3, T4, TG and increased the activity of TPO in a concentration- and time-dependent manner. The peak of T3, T4 and TG secretion was 2 mg / mL and TSAb was 48 h and 2 mg / , 3 mg / m L TSAb for 72 h; the peak of TPO activity was 4 mg / m LTSAb for 48 h. At the same time, the inhibition of cyclic AMPK / PKA pathway by PKA inhibitors reduced the production of T3 by 6.33%, T4 by 6.79%, TG by 51.5%, and the TPO activity by 79.2% (P> 0.05). The intracellular Ca2 + blockade of PIP2 / Ca2 + pathway in BAP-TA-AM cells partially inhibited the generation of T3, T4 and TG, but had no effect on TPO activity The inhibitor of genistein had no significant effect on the activities of T3, T4, TG and TPO (P> 0.05). Conclusion (1) TSAb can