别嘌呤醇对同型半胱氨酸和低密度脂蛋白协同损伤血管内皮细胞的保护作用  

Protective Effect of Allopurinol on Endothelial Cell Injury in Human Umbilical Vein Induced by Homocysteine and Low Density Lipoprotein

作　　者：郑平[1] 刘学文[2] 祁哲[1] 丛祥凤[2] 

机构地区：[1]中国医学科学院中国协和医科大学心血管病研究所阜外心血管病医院冠心病诊治中心,北京市100037 [2]中国医学科学院中国协和医科大学心血管病研究所阜外心血管病医院生化研究室,北京市100037

出　　处：《中国循环杂志》2005年第3期172-175,共4页Chinese Circulation Journal

摘　　要：目的:探讨别嘌呤醇(Allo)对同型半胱氨酸(Hcy)和低密度脂蛋白(LDL)协同损伤人脐静脉血管内皮细胞的保护作用。方法:①将人脐静脉血管内皮细胞与一定浓度的Hcy、LDL及Allo共同培养24小时,分为Hcy(H)组、Hcy+Allo(H+A)组、LDL(L)组、Allo(A)组、LDL+Allo(L+A)组、Hcy+LDL(H+L)组、Hcy+LDL+Allo(H+L+A组,其中Allo在MTT试验中的浓度分别为0.1、0.2、0.3mmol/l)及正常对照组(各n=6孔),应用MTT试验观察Allo是否对其有保护作用。②测定细胞培养上清液乳酸脱氢酶、丙二醛和一氧化氮含量,分别观察内皮细胞损伤、脂质过氧化程度及血管舒张因子产生情况(H+L+A组中Allo为0.2mmol/L)。③同时,利用半定量逆转录聚合酶链式反应方法,检测内皮型一氧化氮合酶转录水平,以观察Allo是否影响其的转录。结果:H+L组、H+A组、H组、H+L+A组(Allo0.1～0.3mmol/L)细胞与正常对照组相比,吸光度减小,分泌一氧化氮减少,乳酸脱氢酶和丙二醛增加,有显著性差异(P<0.05)。除H+L+A组(Allo0.1mmol/L)外,其余各组细胞吸光度较H+L组均明显升高,有显著性差异(P<0.05)。H+L+A组(Allo0.1～0.3mmol/L)细胞吸光度随着Allo浓度增加而增加。除H组外,余各组与H+L组比较一氧化氮生成量均增加,有显著性差异(P<0.05)。与H+L组比较,其余各组细胞损伤、脂质过氧化程度均减轻,有显著性差异(P<0.05)。内皮型一氧化氮合酶的半定量逆转录聚合酶链式反应结果显示各组间的产物电泳带无显著差异。结论:Allo对Hcy和LDL协同损伤人脐静脉血管内皮细胞有保护作用,提示其抗动脉粥样硬化的作用。Objective:To investigate the influence of allopurinol(Allo) on dysfunction induced by homocysteine(Hcy) and low density lipoprotein(LDL)by culturing human umbilical veinendothelium cells(HUVEC). Methods:HUVEC cultured in vitro were used as target cells and were cultured with certain concentrations of Hcy,LDL and Allo.Microculture tetrozolium(MTT)was used to observe the amount of living cells.The content of malondialdehyde and the activity of lactate dehydrogenase were determined to observe the degree of lipid peroxidation and the damage of HUVECs.The amount of NO 2-/NO 3- was determined to observe the capacity of secreting relaxing factor.eNOS mRNA expression was observed by reverse transcriptase-polymerase chain reaction(RT-PCR) to determine whether allopurinol influence transcription of eNOS. Results:The contents of malondialdehyde and lactate dehydrogenase in Hcy(H) group and Hcy+LDL(H+L)group were significantly increased(p<0.05)when compared with the control group,while the MTT optical density(OD),the content of NO 2-/NO 3-were significantly decreased(p<0.05).Malondialdehyde and lactate dehydrogenase in H+L group were significantly increased compared with H group(P<0.05),while the MTT OD was decreased(p<0.05).Malondialdehyde and lactate dehydrogenase in Hcy+LDL+Allo(0.2mmol/L)(H+L+A)group were significantly lower than H+L group(p<0.05),while the MTT OD and NO 2-/NO 3-were significantly higher in the group of H+L+A than in the H+L group(p<0.05).Meanwhile,the MTT OD was significantly increased in an allopurinol-dose-dependent manner.There were no differences in the detections between LDL group and the control group.Allopurinol had no effect on transcription of eNOS. Conclusion:Allopurinol can protect HUVECs against synergistic impairment by homocysteine and LDL,suggesting that allopurinol can inhibit atherosclerosis.

关 键 词：别嘌呤醇 同型半胱氨酸 低密度脂蛋白 

分 类 号：R541[医药卫生—心血管疾病]