刘　倩，胡　芳，牛文彦（天津医科大学免疫学系，天津　300070）



AMPK腺病毒载体在小鼠骨骼肌中的表达和作用

刘倩，胡芳，牛文彦
（天津医科大学免疫学系，天津300070）

摘要目的：探讨注射AMPK腺病毒在小鼠骨骼肌中的表达和作用。方法：C57BL/6小鼠随机分为4组:（1）无任何处理的空白对照组。（2）肌肉注射腺病毒空载体（Ad-GFP）组。（3）肌肉注射Ad-GFP，48 h后腹腔注射AMPK激活剂AICAR组。（4）肌肉注射GFP标记的激活型AMPK腺病毒（Ad-AMPK-CA）组。注射腺病毒72 h后，处死小鼠，通过小动物成像系统测定骨骼肌中的荧光强度，检测腺病毒的表达，用Western blot方法检测ACC的磷酸化。结果：与空白对照组比较，注射腺病毒组的平均荧光强度显著增高。与病毒空载体组比较，注射AICAR组和Ad-AMPK-CA组的ACC磷酸化水平显著升高。结论：AMPK腺病毒能在小鼠骨骼肌中表达目的蛋白，调节AMPK的作用。

关键词AMPK；腺病毒；小鼠；骨骼肌

Effect and expression of AMPK adenovirus on mouse skeletal muscle

LIU Qian,HU Fang,NIU Wen-yan
（Department of Immunology,Tianjin Medical University,Tianjin 300070,China）

Abstract Objective:To explore the effect and expression of injected AMPK adenovirus on mouse skeletal muscle.Methods:Ten weeks old C57BL/6 male mice were randomly divided into four groups.Mice in group1 were normal control group without treatment.Mice in group 2 were intramuscularly injected green fluorescence protein adenovirus(Ad-GFP).Mice in group 3 were intramuscularly injected Ad-GFP,after which were intraperitoneally injected AMPK activator AICAR in 48 h.Mice in group 4 were intramuscularly injected Ad-AMPK-CA.Seventy two hours after adenovirus treatment,all mice were executed and the expression of adenovirus was analysed in skeletal muscle.Fluorescence intensity and ACC phosphorylation in skeletal muscle were analysed by vivo imaging system and western blot,respectively.Results:Compared with the control group,the mean fluorescence intensity of the experimental groups which were injected adenovirus was significantly higher.Compared with the Ad-GFP group,ACC phosphorylation of AICAR and Ad-AMPK-CA groups were significantly increased.Conclusion:AMPK adenovirus can express the target protein in mouse skeletal muscle,and regulate the activity of AMPK.

Key words AMPK；adenovirus；mouse；skeletal muscle

2型糖尿病以葡萄糖代谢异常为特征，主要发病机制是胰岛素抵抗[1-2]，肥胖是其主要诱因。通过非胰岛素依赖机制促进外周组织对葡萄糖的摄取和脂肪酸的氧化是2型糖尿病治疗的关键。单磷酸腺苷（AMP）激活的蛋白激酶（AMPK）是能量代谢和响应外部刺激的一个关键的调节器[3]。它是由一个催化亚基（α）和两个调节亚基（β，γ）组成的异三聚体酶[4]。在营养缺乏和病毒感染时，升高胞内AMP / ATP比值和活化一些上游激酶来激活AMPK，导致α亚基中的苏氨酸172的磷酸化增加，激活AMPK[5]。当AMPK被激活时，通过调节下游的信号通路，例如，AMPK直接磷酸化乙酰-CoA羧化酶（ACC）和HMG-CoA还原酶（HMGCR），能抑制合成代谢，并促进分解代谢，增加骨骼肌对葡萄糖的摄取，增强脂肪酸的氧化作用和胰岛素的敏感性，因此AMPK及其信号通路已成为2型糖尿病的预防和治疗的热点。……