同型半胱氨酸诱导人脐静脉血管内皮细胞活性氧升高的机制研究  

The Mechanisim Underlying Enhancement of ROS Production in Vascular Endothelial Cells Induced by Homocysteine

作　　者：杨红玲 张敏[2] 甘萍[3] 田兴亚[3] 单妍[3] 李树德[4] 

机构地区：[1]云南省第三人民医院心内科,云南昆明650011 [2]昆明医学院第一附属医院心内科,云南昆明650032 [3]昆明医学院海源学院生物化学教研室,云南昆明650031 [4]昆明医学院生物化学教研室,云南昆明650031

出　　处：《昆明医学院学报》2009年第6期19-22,26,共5页Journal of Kunming Medical College

基　　金：国家自然科学基金资助项目(30560036);昆明医学院海源学院科研基金资助项目(JYT0901H)

摘　　要：目的探讨同型半胱氨酸诱导血管内皮细胞活性氧(ROS)升高的机制.方法用DCF-DA荧光探针测定人脐静脉血管内皮细胞(HUEVC)内ROS;用[3H]-胸腺嘧啶脱氧核苷掺入法测定细胞DNA合成率;采用实时定量PCR测定基因的表达.结果同型半胱氨酸诱导HUEVC的ROS水平升高并抑制细胞增殖.抗氧化剂TEMPOL降低同型半胱氨酸对HUEVC增殖的抑制作用.同型半胱氨酸抑制HUEVC的过氧化氢酶和谷胱甘肽氧化还原酶的基因表达.在高同型半胱氨酸血症小鼠胸主动脉中,过氧化氢酶、锰超氧化歧化酶和谷胱甘肽氧化还原酶基因表达均明显降低.结论同型半胱氨酸诱导的ROS产生导致HUEVC增殖抑制.同型半胱氨酸诱导的ROS升高与抗氧化酶基因表达下调有关.Objective To investigate the mechanism underlying the enhancement of homocysteine-indueed production of reactive oxygen species （ROS） in vascular endothelial cells. Methods The levels of ROS in cells were measured by using DCF-DA as a fluorescence probe; DNA synthesis in cells was determined by using [3H] -thymidine incorporation. The expression of tested genes was detected by real-time PCR. Results Homocysteine promoted the levels of ROS and inhibited proliferation in HUEVC. Antioxidant TEMPOL suppressed the inhibitory effect of homoeysteine on proliferation in HUEVC. Homocysteine down-regulated the expression of catalase and glutathione peroxidase. The expression of catalase, glutathione peroxidase, and Mn-SOD decreased in aorta of mice with hyperhomocysteinemia. Conclusion ROS induced by homocysteine inhibit the proliferation of HUEVC. The increase in ROS production induced by homocysteine is due to down-regulation of the expression of antioxidant genes.

关 键 词：同型半胱氨酸 血管内皮细胞 活性氧物质 抗氧化酶 增殖 

分 类 号：R57[医药卫生—消化系统]