摘要
目的:观察具有调脂作用的阿托伐他汀钙在不同浓度下对肿瘤坏死因子诱导体外培养的人脐静脉内皮细胞表达E-选择素和细胞间黏附分子1的影响。方法:①实验于2002-05/2003-12在大连医科大学附属第二医院实验室完成。将健康婴儿脐带进行无菌处理,用D-Hank’s液冲洗脐静脉,1g/L胶原酶消化、离心,置于37℃、体积分数0.05CO2培养箱中培养,选择第2,3代细胞作为实验用。②用肿瘤坏死因子40μg/L、不同浓度(0.005～10μmol/L)的阿托伐他汀钙与体外培养的人脐静脉内皮细胞共同孵育24h后,提取细胞的总RNA,采用半定量反转录聚合酶链反应在mRNA水平进行分析。③用曲线图表示E-选择素和细胞间黏附分子1表达在不同浓度阿托伐他汀钙影响下的变化趋势。结果:①在mRNA水平,阿托伐他汀钙对肿瘤坏死因子诱导的人脐静脉内皮细胞所表达的E-选择素与细胞黏附分子1均具有浓度依赖性。②阿托伐他汀钙对肿瘤坏死因子诱导人脐静脉内皮细胞表达E-选择素mRNA呈促进作用,即阿托伐他汀钙在0～10μmol/L的浓度区间内,随着其浓度由0,0.005,0.01μmol/L渐升至10μmol/L,E-选择素mRNA的表达呈逐渐升高的趋势。③阿托伐他汀钙对肿瘤坏死因子诱导人脐静脉内皮细胞表达细胞间黏附分子1的影响呈双向性,具有浓度差异性。即随着阿托伐他汀钙浓度由0,0.005,0.01μmol/L升至0.05μmol/L,细胞间黏附分子1mRNA表达呈逐渐下降趋势,而在高浓度(0.05～10μmol/L)区间,随着阿托伐他汀钙浓度0.05,0.1μmol/L渐升至10μmol/L,细胞间黏附分子1mRNA的量呈逐渐升高的趋势。结论:在基因水平,阿托伐他汀钙对肿瘤坏死因子诱导血管内皮细胞黏附分子的表达有着确切的调节作用,且存在着浓度依赖性,并在不同的浓度区间,对不同的黏附分子作用不同。
AIM： To observe the influence of atorvastatin calcium, a kind of 3-Hydroxy-3-methylglutaryl coenzyme A （HMG-CoA） reductase inhibitors （statins）, at different concentrations on the expression of E-selectin and intercellular adhesion molecule-1 in cultured human umbilical vein endothelial cells activated by tumor necrosis factor-alpha in vitro.
METHODS： ① The experiment was completed in laboratories of Second Affiliated Hospital, Dalian Medial University from May 2002 to December 2003. Human umbilical veins in healthy neonatal umbilical cords were washed by D-Hank＇s solution, digested by collagenase （1 g/L）. Human umbilcal vein endothelial cells isolated by centrifugation were maintained at 37 ℃ and 5% COy The second and third generations of the above were used in experiments. ② After 24 hours co-incubation of 40 μg/L tumor necrosis factor-alpha, 0.005-10 μmol/L atorvastatin calcium and cultured human umbilical vein endothelial cells in vitro. The total RNA was extracted from the cultured human umbilical vein endothelial cells. The level of mRNA was analyzed by semi-quantitative reverse transcription-pelymerase chain reaction （RT-PCR）. ③ The changes of E-selectin and intercellular adhesion molecule-1 affected by atorvastatin calcium at the different concentration were presented by curve graphs.
RESULTS： ① Atorvastatin calcium had a concentration-dependent effect on the mRNA expression of E- selectin in human umbilcal vein endothelial cells induced by tumor necrosis factor and intercellular adhesion molecule- 1.② The atorvastatin calcium had stimulative effects on the mRNA expression of E- selectin in human umbilcal vein endothelial cells induced by tumor necrosis factor. That was, with the increasing of atorvastatin calcium at 0-10μmol/L concentration level, the concentration from 0,0.005, 0.01 μmol/L decreased to 10 μmol/L. The mRNA expression of E-selectin was enhanced gradually. ③ Effect of atorvastatin calcium on intercellular adhesion molecule-1 in human umbilcal vein endothelial cells induced by tumor necrosis factor was bi-directional with concentration difference. That was, with the increase of atorvastatin calcium from 0, 0.005, 0.01 μmol/L increased to 0.05 μmol/L. The mRNA expression of intercellular adhesion molecule-1 was reduced gradually. While with the increase of atorvastatin calcium from 0.05,0.1 μmol/L increased to 10 μmol/L, the mRNA expression of intercellular adhesion molecule-1 were enhanced at higher concentrations （0.05 to 10 μmol/L）.
CONCLUSION： The atorvastatin calcium has exactly adjustable effects on the expression of adhesion molecules in cultured human umbilical vein endothelial cells induced by tumor necrosis factor with concentration-dependent effects, and the effects are different between the different adhesion molecules at different concentrations.
出处
《中国临床康复》
CSCD
北大核心
2005年第46期56-59,共4页
Chinese Journal of Clinical Rehabilitation
基金
国家自然科学基金资助项目(30271433)~~
