中文摘要：

目的：探讨促甲状腺激素（ TSH）调节肝脏AMP激活的蛋白激酶（ AMPK）α亚基的多磷酸化位点，揭示TSH调节AMPKα活性可能存在的机制。方法①在TSH刺激HepG2细胞组和对照组、TSH受体敲除（ Tshr-ko）和同窝配对野生型（ WT）小鼠中，用实时定量 PCR和 Western blotting 检测AMPKαmRNA、总蛋白、苏氨酸（ Thr）172和丝氨酸（ Ser）173、乙酰辅酶A羧化酶（ ACC） Ser 79磷酸化水平；②AMPK激活剂AICAR、PKA抑制剂H89或TSH刺激肝细胞，检测AMPK、ACC磷酸化水平。结果相对于对照组，TSH刺激HepG2细胞后， AMPKα总蛋白水平无统计学差异，AMPKαThr 172磷酸化减少（P＜0．05），Ser 173表达无明显变化（P＞0．05）。与WT小鼠比较，Tshr-ko小鼠肝脏AMPKαmRNA和总蛋白水平表达不变，AMPKαThr 172（P＜0．05）和ACC Ser 79（P＜0．01）磷酸化升高，Ser 173磷酸化无改变（P＞0．05）。 AICAR和H89增加AMPKαThr 172和ACC Ser 79的磷酸化。 H89与TSH共刺激HepG2细胞时， TSH减少AMPK αThr 172蛋白表达的作用被部分反转。结论 TSH通过TSH受体调节肝脏AMPKαThr 172而非Ser 173的磷酸化。

英文摘要：

Objective To explore the effect of thyroid-stimulating hormone （ TSH） on the multisite phosphorylation of hepatic AMP-activated protein kinase （ AMPK）αsubunit, and to reveal the possible mechanism of TSH-regulated AMPKαactivity in the liver.Methods ①In the HepG2 cells stimulated with or without TSH, and the TSH receptor knockout （ Tshr-ko） mice and their littermate mice, the expressions of hepatic AMPKαmRNA, total AMPKαprotein, phosphorylation of AMPKαThr 172, AMPKαSer 173 and acetyl-CoA carboxylase （ ACC） Ser 79 were detected by re-al-time PCR and Western blotting.② With AICAR （AMPK activator）, H89 （PKA inhibitor） or TSH stimulation, AMPKαand ACC phosphorylation were tested in liver cells.Results Compared to the control group, TSH decreased＆nbsp;phosphorylation of AMPKαThr 172 （ P0.05）.Compared to WT mice, the expressions of AMPKαmRNA and total protein remained unchanged in Tshr-ko mouse liver.The phosphorylation of AMPKαThr 172 （P0.05）.AICAR and H89 increased phosphorylation of AMPKαThr 172 and ACC Ser 79.When H89 was employed, the TSH-induced decrease of AMPKαThr 172 phosphorylation was partly reversed.Conclusion TSH reg-ulates the phosphorylation of hepatic AMPKαThr 172 instead of Ser 173 by acting on thyrotropin receptor.