摘要
目的观察HMG-COA还原酶抑制剂—辛伐他汀对apoE基因缺陷小鼠早期氧化应激及主动脉内皮微膜蛋白-1的影响,探讨辛伐他汀在动脉粥样硬化的一级预防中保护内皮功能的机制。方法24只6周龄雄性apoE基因缺陷小鼠,随机等分为两组:对照组、辛伐他汀(5 mg/(kg·d))治疗组,4周后处死动物。分光光度法测定血总胆固醇、超氧化物歧化酶活性、丙二醛和一氧化氮含量;采用HE染色法观察小鼠主动脉内皮组织;免疫组织化学技术分析小鼠主动脉内皮的微膜蛋白-1的表达。结果治疗4周时治疗组与对照组比较,两组间血胆固醇水平无显著性差异(11.8963±0.2036 mmol/L比对照组:10.8014±2.0984 mmol/L,P＞0.05);治疗4周时与对照组相比,辛伐他汀治疗组血清超氧化物歧化酶活性(135.3897±5.5664 u/L比对照组:97.2866±7.6414 u/L,P＜0.01)和一氧化氮水平(28.4946±4.1529 umol/L比对照组:12.3656±2.1816 umol/L,P＜0.01)均明显升高;治疗4周时治疗组与对照组比较,丙二醛水平显著减低(10.5641±0.5941 nmol/L比对照组:17.3846±1.0562 nmol/L,P＜0.01);治疗4周时与对照组比较,辛伐他汀治疗组主动脉内皮组织的损伤减轻(损伤阳性率33.33%比对照组:75.00%,P＜0.05);治疗4周时与对照组比较,辛伐他汀治疗组主动脉内皮上微膜蛋白-1的表达明显降低(表达阳性率41.67%比对照组:83.33%,P＜0.05)。结论辛伐他汀在不影响血胆固醇水平情况下,抑制主动脉内皮的微膜蛋白-1的表达,减轻apoE基因缺陷小鼠氧化应激,增加NO水平,起到改善内皮功能、抗动脉粥样硬化作用,在动脉粥样硬化的一级预防中可能发挥重要作用。
Objective: To investigate the mechanisms by which Simvastatin, one kind of 3-hydroxy-3-methylglutaryl coenzyme A(HMG-CoA) reductase inhibitor, plays an important role in primary prevention of atherosclerosis independently of its lipid-lowering effect in Apolipoprotein E -deficient mice in the early stage of atherosclerosis. Methods : Twenty-four 6-week old male apoE-deficient mice were randomly divided into two groups: control group(normal saline) and treatment group(simvastatin (5 mg/(kg.d)). Simvastatin was administered to treatment group mice by gavage and the same volume of normal saline was administered to control group mice by the same method for 4 weeks. Total cholesterol (TC), super-oxide dismutase (SOD), malondialdehyde (MDA) and serum nitric oxide (NO) were measured by spectrophotometer analysis. Endothelium tissue was observed by HE dyeing; The expression of caveolin-1 in aortic wall was detected by immunohistochemistry. Results: There was no significant difference in serum TC (U.8963±0.2036 mmol/L vs control's :10.8014±2.0984 mmol/L,P >0.05, at 4-week) between control and treatment groups. Compared with the control's, the effects of simvastatin were more significant in decreasing serum MDA level(10.5641±0.5941 nmol/l vs control's: 17.3846±1.0562 nmol/l,P<0.01,at 4-week), increasing serum SOD level (135.3897±5.5664 u/l vs control's : 97.2866±7.6414 u/l,P<0.01 , at 4-week) and NO level (28.4946±4.1529 umol/l vs control's: 12.3656±2.1816 umol/1, P <0.01 , at 4-week)at 4 weeks. More damaged endothelium was discovered in control group than simvastatin treatment group(33.33%vs control's :75.00%, P <0.05 ,at 4-week). Expression of caveolin-1 was significantly inhibited in the simvastatin treatment group(41.67%vs control's: 83.33%, P <0.05, at 4-week) than in control group. Conclusions: Simvastatin attenuates oxidative stress and protects endothelial function by the mechanisms of downregulating of caveolin-1 expression , decreasing serum MDA level in aortic wall , increasing serum SOD level and NO level, which were inconsistent with its cholesterol-lowering effect. It may play an important role in primary prevention of atherosclerosis and might be independent of lipid-regulation mechanism , if not all.
引文
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[10] Dever G, Spickett CM, Kennedy S, et al. The nitric oxide-donating pravastatin derivative,NCX6550-1,2,6,7,8,8a-Hexahydro-beta,delta,6-trihydroxy-2-m ethyl-8-(2-methyl-1 -oxobutoxy)-1 -naphtalene-heptanoic ac(?)d 4-(nitrooxy) butyl ester)], reduces splenocyte adhesion and reactive oxygen species generation in normal and atherosclerotic mice[J]. J Pharmacol Exp Ther, 2007,320:419-426.
[11]Sethy-Coraci I,Crock LW,Silverstein SC.PAF-receptor antagonists,lovastatin,and the PTK inhibitor genistein inhibit H_2O_2 secretion by macrophages cultured on oxidized-LDL matrices[J].J Leukoc Biol,2005,78:1166-1174.
[12]Kowalski J,Pawlicki L,Grycewicz J,et al.Estimation of antioxidative effect of atorvastatin and fluvastatin used in primary prevention of coronary heart disease—effect on antioxidative enzymatic activity[J].Wiad Lek,2005,58:275-279.
[13]Lim EJ,Smart EJ,Toborek M,et al.The role of caveolin-1 in PCB77-induced eNOS phosphorylation in human-derived endothelial cells[J].Am J Physiol Heart Circ Physiol,2007,293:H3340-3347.
[14]Vera R,Sanchez M,Galisteo M,et al.Chronic administration of genistein improves endothelial dysfunction in spontaneously hypertensive rats:involvement of eNOS,caveolin and calmodulin expression and NADPH oxidase activity[J].Clin Sci,2007,112:183-191.
[15]Maniatis NA,Brovkovych V,Allen SE,et al.Novel mechanism of endothelial nitric oxide synthase activation mediated by caveolae internalization in endothelial cells[J].Circ Res,2006,99:870-877.
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