缺氧/复氧损伤人脐静脉内皮细胞黏附分子表达与新型植物雌激素α-玉米赤霉醇的影响  

Influence of a new type of phytoestrogen alpha-zearalanol on the expression of cell adhesion molecule in human umbilical vein endothelial cells following hypoxia/reoxygenation injury

作　　者：王雯[1] 蒋东桥[1] 邱笑违[1] 朱盈芬[1] 张立克[1] 

机构地区：[1]首都医科大学基础医学院病理生理学教研室,北京市100069

出　　处：《中国组织工程研究与临床康复》2007年第6期1038-1040,共3页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：北京市优秀人才培养专项经费(20042D0501817);首都医科大学科研基金(2004ZR02)~~

摘　　要：目的:探讨新型植物雌激素α-玉米赤霉醇对缺氧/复氧损伤后人脐静脉内皮细胞黏附分子表达的干预,并与内源性动物雌激素雌二醇的作用进行比较。方法:实验于2005-10/2006-01在首都医科大学完成。①健康产妇的婴儿脐带由首都医科大学附属宣武医院妇产科提供,产妇及其家属签署捐赠同意书。α-玉米赤霉醇(中国农业大学提纯,纯度99%以上,以乙醇作为溶剂),雌二醇(Sigma公司)。②脐静脉内皮细胞原代培养后,用质量浓度为1.25g/L的胰蛋白酶+0.1g/L的乙二胺四乙酸混合消化液进行传代,传至2～3代后用于实验。细胞以3×105/孔接种于24孔培养板,待生长至80%～90%融合后,设立10组:正常对照组、模型对照组、α-玉米赤霉醇1,10,100,1000nmol/L组、雌二醇1,10,100,1000nmol/L组,6孔/组。③除正常对照组外,其余各组细胞均复制缺氧/复氧损伤模型,置于体积分数为0.93的N2+0.05的CO2+0.02的O2缺氧环境中3h,然后恢复正常氧供应1h。α-玉米赤霉醇各组于造模前20min分别加入α-玉米赤霉醇,使细胞培养液中α-玉米赤霉醇终浓度分别达到1,10,100,1000nmol/L;雌二醇各组于造模前20min分别加入雌二醇,使细胞培养液中雌二醇终浓度分别达到1,10,100,1000nmol/L;正常对照组、模型对照组不予任何药物处理。④吸取各组上清液,按酶联免疫吸附试剂盒说明操作分别测定各孔可溶性E-选择素、细胞间黏附分子1、血管细胞黏附分子1的含量。结果:①缺氧/复氧后各组细胞E-选择素含量的比较:与正常对照组比较,缺氧3h/复氧1h后模型对照组脐静脉内皮细胞上清液中的E-选择素含量明显升高[(1.77±0.36),(5.62±0.74)pg/L,P<0.01]。与模型对照组比较,α-玉米赤霉醇1,10,100,1000nmol/L组E-选择素含量均明显降低[(5.62±0.74),(3.53±1.21),(3.16±0.94),(2.79±1.78),(2.18±0.75)pg/L,P<0.05或0.01],雌二醇1,10,100,1000nmol/L组E-选择素含量亦均明显降低[(5.62±0.74),(3.72AIM： To explore the interventional effect of new phytoestrogen α-zearalanol on hypoxia/reoxygenation （H/R） induced cell adhesion molecule expression in human umbilical vein endothelial cells （HUVECs） and compare this effect with endogenous estrogen. METHODS： The experiment was done in Capital Medical University from October 2005 to January 2006. （1） Infant umbilical cores of healthy parturients were supplied by Department of Gynaecology and Obstetrics, Xuanwu Hospital of Capital Medical University, with consent of puerperant and their families. α-zearalanol was purified by Chinese Agriculture University, which was solved in ethanol, purity was over 99%. Estradiol was supplied by Sigma Corporation. （2）After primary culture, HUVECs were passaged with 1.25 g/L trypsin and 0.1 g/L ethylenediamine tetraacetic acid （EDTA）. Cells after 2-3 generations were used in subsequent experiments. HUVECs were inoculated in 24-well plates with 3×10^5/well. After 80%-90% confluence, HUVECs were divided into ten groups： normal control group, model control group, 1,10,100,1 000 nmol/L α-zearalanol groups, 1,10,100,1 000 nmol/L estradiol groups. There were six wells in every group. （3）Except normal control group, other HUVECs were all treated with H/R injury. That was, cultured in hypoxia environment （0.93 N2＋0.05 CO2＋0.02 O2） for 3 hours, and then recovered to normal environment for 1 hour, Twenty minutes before hypoxia, the α-zearalanol groups received α-zearalanol in culture fluid in order to their last concentrations of 1,10,100,1 000 nmol/L; the estradiol groups received estradiol in culture fluid in order to their last concentrations of 1,10,100,1 000 nmol/L. There was no drug treatment either in normal control group or model control group. （4）The levels of E-selectin, intercellular adhesion molecule-1 （ICAM -1 ）and vascular cell adhesion molecule-1 （VCAM-1） in supematant of HUVECs was detected by enzyme-linked immunosorbent assay （ELISA）. RESULTS： （1）Comparis

关 键 词：植物雌激素类 雌二醇 内皮 血管 脐静脉 缺氧 细胞粘附分子 

分 类 号：R285.5[医药卫生—中药学]