AMPK腺病毒载体在小鼠骨骼肌中的表达和作用

摘要

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.%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。