别嘌呤醇对血管内皮细胞舒张功能的保护作用  

Protective Effect of Allopurinol on Endothelial Cell Injury in Human Umbilical Vein

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阜外心血管病医院冠心病诊治中心,100037 [2]中国医学科学院中国协和医科大学阜外心血管病医院生化检验中心,100037

出　　处：《中国分子心脏病学杂志》2005年第1期405-409,共5页Molecular Cardiology of China

基　　金：北京人事部回国人员资金(38)

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(HUVEC)舒张功能的保护作用。方法将HUVEC与一定浓度的Hey、LDL及不同浓度的hllo共同培养24h,应用MTT试验(用甲欣光吸收OD值表示活细胞数)观察Allo是否对HUVEC有保护作用;测定细胞培养上清液一氧化氮(NO)含量,观察HUVEC血管舒张因子产生情况。同时,利用半定量RT-PCR方法测定内皮型一氧化氮合酶(eNOS)转录水平,以观察Allo是否影响eNOS的转录。结果HUVEC培养24h后,Hey(H)组、Hcy+LDL(H+L)组与Con组相比,活细胞数减少,NO产生减少(P值均<0.05)。H+L组与H组相比,活细胞数进一步减少(MTT试验OD值0.179±0.004.vs0.217±0.001)。Hcy+LDL+Allo(0.2mmol/L)(H+L+A)组,与H+L组相比,活细胞数增加(MTY试验OD值0.207±0.002),NO增加(15.58±0.193μmol/lvs14.21±0.340μmol/1)(P值均<0.05)。同时,活细胞数随Allo浓度增加而升高,至0.3mmol/L时趋于稳定。eNOS的半定量RT-PCR结果显示各组间的产物电泳带无明显差异。结论(1)Hcy可导致内皮细胞损伤、功能障碍;(2)Hcy和LDL对内皮细胞的损伤和功能障碍具有协同作用;(3)别嘌呤醇对Hcy和LDL协同损伤HUVE有保护作用。提示别嘌呤醇对体内存在高氧化状态的患者有对抗动脉粥样硬化的作用。Objective To study the impact of homocysteine （Hcy） and low density lipoprotein （LDL） on endothelial cell injury, and to observe whether allopurinol （Allo） can prevent the dysfunction induced by them. Methods Human umbilical vein endothelial cells were used as target cells and were cultured with certain concentration of Hcy, LDL and Allo. MTT was used to observe the amount of living cells. The amount of NO^2-/NO^3- were determined to observe the capacity of secreting relaxing factor, eNOS mRNA expression was observed by RT - PCR to determine whether allopurinol influenced transcription of eNOS. Results The MTT optical density （OD）, the content of NO^2-/NO^3- of group H and group H ＋ L significandy decreased （P 〈0.05）. The MTT OD （0.207 ±0. 002） and the content of NO^2-/NO^3- （15.58 ±0. 193 μmol/L vs 14.21 ＋ 0. 340μmol/L） of Group H ＋ L ＋ A （0.2 mmol/L） was significantly higher than Group H ＋ L （ P 〈 0.05 ）. Meanwhile, the MTT OD increased significantly in an allopurinol - dose - dependent manner. Allopurinol had no impact on transcription of eNOS. Conclusion （ 1 ） Hcy can induce HUVECs＇ injury; （2） Hcy and LDL had synergism impairment effect on HUVECs; （3）Allopurinol had protective effects against the synergism impairment impact of Hcy and LDL on HUVECs. It may suggest that allopurinol can inhibit atherosclerosis in those people whose body exists increasing oxidative stress.

关 键 词：别嘌呤醇 同型半胱氨酸 低密度脂蛋白 内皮细胞 动脉粥样硬化 

分 类 号：R543[医药卫生—心血管疾病]