血管紧张素Ⅱ和坎地沙坦对小鼠骨骼肌细胞C2C12胰岛素敏感性的影响  

Effects of Angiotensin Ⅱ and Candesartan on insulin sensitivity of mouse C2C12 myotubes

作　　者：李凡[1] 谢静[1] 英明中[1] 曾强[1] 

机构地区：[1]解放军总医院国际医学中心,北京100853

出　　处：《中国医药导报》2015年第7期14-18,共5页China Medical Herald

基　　金：北京市自然科学基金项目(编号7122171)

摘　　要：目的研究血管紧张素Ⅱ和坎地沙坦对小鼠骨骼肌细胞(C2C12)胰岛素敏感性的影响及其机制。方法 C2C12诱导分化成熟后,分为对照组(C组)、模型组(M组,AngⅡ0.1μmol/L)、坎地沙坦低剂量组(Can1组,坎地沙坦0.1μmol/L+AngⅡ0.1μmol/L)、坎地沙坦中剂量组(Can2组,坎地沙坦1μmol/L+AngⅡ0.1μmol/L)、坎地沙坦高剂量组(Can3组,坎地沙坦10μmol/L+AngⅡ0.1μmol/L)。各组细胞给予相应药物及胰岛素处理后,使用二氯荧光素二乙酸酯(DCFH-DA)检测细胞内活性氧(ROS)的生成水平,使用2-脱氧葡萄糖(2-NBDG)检测细胞对胰岛素的敏感性,并用RT-PCR法和Western印迹法分别检测各组细胞内的核转录因子-2(Nrf2)的m RNA及蛋白表达水平。结果与C组比较,M组ROS生成水平显著升高(P<0.01),摄取2-NBDG量显著降低(P<0.01);与M组比较,Can3组ROS生成水平显著减少(P<0.05),摄取2-NBDG量则显著增加(P<0.05)。M组Nrf2的m RNA及蛋白表达较C组显著降低(P<0.01),Can2组及Can3组Nrf2的m RNA及蛋白表达较M组显著升高(P<0.01或P<0.05)。结论 AngⅡ通过下调C2C12中Nrf2的m RNA及蛋白表达,抑制Nrf2的抗氧化效应,增加ROS生成,引起骨骼肌细胞胰岛素抵抗;而坎地沙坦通过抑制AngⅡ的这种作用改善骨骼肌胰岛素抵抗。Objective To investigate the effects of Candesartan and Angiotensin Ⅱ（Ang Ⅱ） on insulin sensitivity of mouse C2C12 myotubes, and discuss the possible mechanism. Methods Mature C2C12 myotubes were divided into 4groups： control group（group C）, model group（group M, Ang Ⅱ 0.1 μmol/L）, low dose Candesartan group（Can1 group,Candesartan 0.1 μmol/L＋AngⅡ 0.1 μmol/L）, mild dose Candesartan group（Can2 group, Candesartan 1 μmol/L＋AngⅡ0.1 μmol/L） and high dose Candesartan group（Can3 group, Candesartan 10 μmol/L＋AngⅡ 0.1 μmol/L）. After the intervention of angiotensin Ⅱ and insulin, the uptake of 2-deoxyglucose（2-NBDG） by C2C12 myotubes was measured in each group as well as the level of reactive oxygen species（ROS）. The m RNA and protein expression of Nrf2 were measured by RT-PCR and Western blotting assay. Results Compared with the group C, the uptake of 2-NBDG in the group M was significantly decreased（P〈0.01）, the m RNA and protein expression of Nrf2 in group M was significantly decreased（P〈0.01）, at the same time, the level of ROS was increased significantly（P〈0.01）. Compared with group M, the uptake of 2-NBGD was increased obviously in Can3 group（P〈0.05）. The m RNA and protein expression of Nrf2 in group M were decreased than those in group C（P〈0.01）, the m RNA and protein expression of Nrf2 in the Can2 group and Can3 group were significantly increased than those in group M（P〈0.01 or P〈0.05）. Conclusion Angiotensin Ⅱ can cause the insulin resistance of the skeletal muscle cells by reducing the m RNA and protein expression of Nrf2, controling Nrf2 oxidation effect, increasing ROS produce; Candesartan can improve skeletal muscle cells insulin resistance by reverse the effect of AngⅡ on Nrf2 and improve the overload of ROS in the cells.

关 键 词：坎地沙坦 血管紧张素Ⅱ 胰岛素抵抗 核转录因子-2 活性氧 

分 类 号：R544.1[医药卫生—心血管疾病]