缬沙坦抑制血管紧张素Ⅱ诱导的血管平滑肌细胞凋亡  

Valsartan inhibits angiotensin Ⅱ-induced vascular smooth muscle cell apoptosis

作　　者：顾建钦[1] 秦历杰[1] 

机构地区：[1]河南省人民医院急诊科,郑州450003

出　　处：《中华急诊医学杂志》2017年第8期895-900,共6页Chinese Journal of Emergency Medicine

基　　金：河南省科技攻关计划项目（122102310589）

摘　　要：目的 探讨缬沙坦对血管紧张素II诱导的血管平滑肌细胞凋亡的影响及其机制。方法 将大鼠胸主动脉平滑肌细胞（A7r5）分为五组：①对照（DMSO）组、②血管紧张素II（angiotensin II, Ang II）100μM组、③血管紧张素II 100 μM ＋缬沙坦（valsartan, Val）10μM组、④血管紧张素II 100μM ＋缬沙坦 10μM＋复合物C（Compoud C, CompC）1μM组、⑤血管紧张素II 100μM＋5-氨基咪哇-4-甲酞胺核糖核普酸（5-Aminoimidazole-4earboxamide-ribo-nucle-oside, AICAR）100μM组，各组细胞予以相应药物处理24后，用分光光度计法检测细胞内Caspase 3活性；流式细胞术检测细胞凋亡率；蛋白印迹法检测细胞内AMPK总蛋白及磷酸化水平的变化；免疫荧光法检测细胞内活性氧（ROS）的含量；WST-1法检测细胞内总超氧化物歧化酶（SOD）活性；TBA法检测细胞内丙二醛（MDA）活性。结果 与对照组相比，AngII组细胞凋亡率增加（P〈0.01），细胞内ROS的生成和Caspase 3、MDA活性增加（P〈0.001），细胞内AMPK的磷酸化水平、SOD活性降低（P〈0.01）；与AngII组相比，AngII＋Val组和AngII＋AICAR组细胞凋亡率降低，（P〈0.05），细胞内ROS的生成和Caspase 3、MDA活性降低（P〈0.001），细胞内AMPK的磷酸化水平、SOD活性增加（P〈0.05）；与AngII＋Val组相比，AngII＋Val＋CompC组细胞凋亡率增加（P〈0.05），细胞内ROS的生成和Caspase 3、MDA活性增加（P〈0.05），细胞内AMPK的磷酸化水平、SOD活性下降（P〈0.01）。结论 缬沙坦可以抑制血管紧张素II诱导的血管平滑肌细胞凋亡，其机制可能与调节细胞内AMPK的磷酸化水平和ROS、SOD、MDA的活性相关。Objective To investigate the impacts of valsartan on cell apoptosis induced by angiotensin Ⅱ in vascular smooth muscle cells, and discuss whether the mechanism is relevant to AMP- Activated Protein Kinases. Methods Vascular smooth muscle cells （ATr5） were designated to 5 groups： （1）control （DMSO） group, （2）Angiotensin Ⅱ （Ang Ⅱ ） 100 μmol/L group, （3）Angiotensin Ⅱ 100 μmol/L ± valsartan 10 μmol/L group, @Angiotensin Ⅱ 100 μmol/L ± valsartan 10 μmol/L ± compound C 1μmol/L group, @ Angiotensin Ⅱ 100 μmol/L ± 5-Aminoimidazole-4earboxamide-ribo-nucle-oside （AICAR） 100 Ixmol/L group, after 24h incubation, the intracellular activity of Caspase 3 was measured by spectrophotometry, the cell apoptosis were enumerated by low cytometry, the intracellular AMP-Activated Protein Kinases （AMPK） phosphorylation and total expression quantity were examined by western blot, the intracellular reactive oxygen species （ROS） was measured by fluorescent probe DCFH-DA, the intracellular activity of total superoxide dismutase （SOD） was measured by WST-1 method, the intracellular activity of Malondialdehyde （MDA） was measured by TBA method. Two groups were compared by using Student t test. Differences among multiple groups were evaluated by ANOVA. Results Compared with control group, the cell apoptosis of Angiotensin Ⅱ group was increased [ （45.46 ± 15.40）% vs. （ 1.88 ± 3.28）%, P = 0.002], the synthesis of ROS was increased [ （9.24 ±0.46） vs. （1.00 ±0.00）, P〈0.01], the activity of Caspase3 was increased [ （35.03±3.54） vs. （13.33±1.79）, P〈0.01], the activity of M DA was increased [ （4. 32 ± 0.73 ） vs. （ 2. 05 ± 0. 18 ） , P 〈 0. 01 ） ] , the phosphorylation of AMPK was decreased, the activity of SOD was decreased [ （90. 29 ± 14. 73） vs. （ 136. 02 ± 18. 82）, P =0. 001 ] ; compared with Angiotensin 1I group, the cell apoptosis of Angiotensin 1[ ± valsartan group and Angiotensin lI ± AICAR group were decreased [ （2

关 键 词：缬沙坦 血管紧张素Ⅱ 血管平滑肌细胞 凋亡 腺苷酸活化蛋白激酶 活性氧 总超氧化物歧化酶 丙二醛 

分 类 号：R96[医药卫生—药理学]