摘要：目的 观察不同时期给予抵挡汤干预对糖尿病大鼠血管内皮细胞腺苷酸活化蛋白激酶（AMPK）信号通路相关因子的变化，探讨其调控AMPK信号通路介导的线粒体能量代谢对糖尿病大血管病变防御功能的作用机制。方法 采用尾静脉注射链脲佐菌素配合高脂饲料喂养制作糖尿病大鼠模型。实验大鼠随机分为正常组、模型组、二甲双胍组、辛伐他汀组和抵挡汤早、中、晚期组，Western blot检测胸主动脉内皮细胞AMPKα1、过氧化物酶体增殖活化受体γ共激活因子-1α（PGC-1α）的蛋白表达，ELISA检测细胞内一磷酸腺苷（AMP）、三磷酸腺苷（ATP）水平，实时荧光定量PCR检测胸主动脉组织Caspase-3、内皮型一氧化氮合酶（eNOS）、Bcl-2的基因表达。结果 与模型组比较，抵挡汤早、中期组和辛伐他汀组大鼠胸主动脉AMPKα1、PGC-1α蛋白表达明显升高（P<0.05）；抵挡汤早期组和辛伐他汀组Bcl-2、eNOS基因表达明显升高（P<0.05），Caspase-3基因表达明显降低（P<0.05）；抵挡汤早期组和辛伐他汀组ATP水平明显升高（P<0.05），AMP水平明显降低（P<0.05），其中以抵挡汤早期组效果最显著。结论 抵挡汤早期干预可通过调控AMPK信号通路，增强血管内皮细胞线粒体的能量代谢，从而加强血管内皮的防御功能。
关键词：抵挡汤；腺苷酸活化蛋白激酶；Caspase-3；能量代谢；早期干预；大鼠
DOI：10.3969/j.issn.1005-5304.2016.10.017
中图分类号：R285.5 文献标识码：A 文章编号：1005-5304（2016）10-0072-06
Abstract： Objective To observe the effects of the intervention of Didang Decoction at different times on changes of AMPK signaling pathway related factors in macrovascular endotheliocytes of diabetic rats； To discuss the mechanism of mitochondria energy metabolism regulating the AMPK signaling pathway for macrovascular endothelial defense function. Methods Injection of STZ into the caudal vein and administration of high fat diet wer used to generate diabetic rat model. All rats were randomly divided into the following 7 groups： control， model， metformin， simvastatin， early-， middle-， and late-stage Didang Decoction group. Western blot was used to detect the expressions of APMKα1 and PGC-1α in rat aortic endothelial cells. Changes in the intracellular AMP and ATP levels were detected by ELISA. Real time fluorescence quantitative PCR was used to detected mRNA expressions of Caspase-3， eNOS， and Bcl-2 in tissue of thoracic aorta. Results Compared with the model group， the expressions of AMPKα1 and PGC-1α in the early-stage and middle-stage Didang Decoction group and simvastatin group increased （P<0.05）； the gene expressions of Bcl-2， and eNOS significantly increased in the early-stage Didang Decoction group and simvastatin group （P<0.05）， while the expressions of Caspase-3 decreased significantly （P<0.05）. The expression of ATP increased significantly and the expression of AMP decreased significantly in the early-stage Didang Decoction group and simvastatin group （P<0.05）， and the best effects were shown in the early-stage Didang Decoction group. Conclusion Early intervention of Didang Decoction can enhance energy metabolism in the mitochondria of macrovascular endothelial cells by regulating the AMPK signaling pathway， and then plays a role in strengthening the defense function of macrovascular endothelial cells.