雌激素对人脐静脉血管内皮细胞凋亡的影响  

Effects of estrogen on the apoptosis of human umbilical vein endothelial cells

作　　者：何小静[1,2] 王彦洁[3] 王宇[4] 杨欣[1] 李晓冬[4] 

机构地区：[1]北京大学人民医院妇科,北京100044 [2]河北医科大学第二医院 [3]北京大学第三医院妇科,北京100000 [4]河北医科大学第二医院妇科,石家庄050000

出　　处：《医学研究生学报》2015年第9期915-919,共5页Journal of Medical Postgraduates

基　　金：国家自然科学基金(81270667)

摘　　要：目的目前认为内质网是心血管疾病治疗的新靶点,其应激反应在高血压心血管重塑中具有重要作用。文中通过建立衣霉素及二硫苏糖醇诱导人脐静脉血管内皮细胞(human umbilical vein endothelial cells,HUVECs)内质网应激(endoplasmic reticulum stress,ERS)凋亡模型,探讨17-β雌二醇对衣霉素/二硫苏糖醇诱导HUVECs ERS凋亡的影响。方法实验前期已确定10μmol/L衣霉素处理细胞10 h或2 mmol/L二硫苏糖醇处理细胞8 h为诱导细胞ERS凋亡最佳浓度和时间。浓度为10-8mol/L的17-β雌二醇对ERS的保护作用最明显。根据是否用衣霉素/二硫苏糖醇,17-β雌二醇,雌激素受体拮抗剂(ICI182,780和G15)处理进行如下分组:1空白对照A、B组;2衣霉素/二硫苏糖醇组;3衣霉素/二硫苏糖醇+17-β雌二醇组;4衣霉素/二硫苏糖醇+17-β雌二醇+ICI组;5衣霉素/二硫苏糖醇+17-β雌二醇+G15组;6衣霉素/二硫苏糖醇+17-β雌二醇+ICI+G15组。Western blot检测ERS诱导凋亡的标志分子GRP78、剪切的caspase 12及CHOP的表达,及Hochest染色检测凋亡细胞数的变化。结果与空白对照A、B组比较,衣霉素/二硫苏糖醇组GRP78、caspase-12和CHOP的表达升高(P<0.05);与衣霉素组比较,衣霉素+17-β雌二醇组GRP78、caspase-12和CHOP的表达均下降(6.80±1.07 vs4.01±0.46,1.54±0.32 vs 0.88±0.10,1.91±0.37 vs 0.91±0.02,P<0.05);与衣霉素+17-β雌二醇组比较,衣霉素+17-β雌二醇+ICI组CHOP表达升高(0.91±0.02 vs 0.96±0.02,P<0.05),衣霉素+17-β雌二醇+G15组GRP78、caspase-12表达升高(4.01±0.46 vs 5.25±0.80,0.88±0.10 vs 1.02±0.07,P<0.05),衣霉素+17-β雌二醇+ICI+G15组GRP78、caspase-12和CHOP的表达升高(P<0.05)。与二硫苏糖醇组比较,二硫苏糖醇+17-β雌二醇组GRP78和CHOP表达均下降(1.81±0.08 vs 1.30±0.14,1.00±0.13 vs 0.51±0.01,P<0.05);与二硫苏糖醇+17-β雌二醇组比较,二硫苏糖醇+17-β雌二醇+ICI组GRP78和CHOP的表达下降(P<0.05),二硫苏糖醇+17-β雌二醇+G15组GRObjective Recent researches find that endoplasmic reticulum is a new target for the treatment of cardiovascular disease and its stress response plays an important role in the hypertension-induced cardiovascular remodeling. The study built up the apoptosis model of endoplasmic reticulum stress （ERS） in human umbilical vein endothelial cells （HUVECs） and investigated the impact of 1713-estradiol （ E2 ） on the ERS apoptosis of HUVECs induced by tunicamycinand （TM）/ dithiothreitol （DTT）. Methods HUVECs incubated for 10 h in 10 μmol/L TM or 8 h in 2 mmol/L UIT were the best concentration and time for ERS apoptosis. E2 of 10^-8 mol/L was prominent in protecting ERS. According to the application of TM/DTT, E2, estrogen antagonists （ICI182, 780 and G15）, there were 6 groups： control group, TM/DTT group, TM/DTT ＋ E2 group, TM/ DTT E2 ＋G15 group, TM/DTT ＋E2 ＋ICI ＋G15 group. Western blot was applied to detect the expressions of biomarkers for ERS induced apoptosis （ GRP78, cleaved caspase-12 and CHOP）. Hochest staining was used to observe the changes of the apoptosis cell number. Results Compared with control group, the expressions of GRP78, caspase-12 and CHOP significantly increased（P 〈0.05）. In comparison with TM group, the expressions of GRP78, caspase-12 and CHOP in TM ＋ E2 group significantly decreased （6.80 ± 1.07 vs 4.01±0.46, 1.54 ±0.32 vs 0. 88 ±0.10, 1.91±0.37 vs 0. 91 ± 0.02, P〈0.05）. In comparison with TM ＋ E2group, the CHOP expression in TM ＋ E2 ＋ ICI group increased（0.91 ±0.02 vs 0.96 ± 0.02, P 〈 0.05 ）, and the expressions of GRP78 and caspase-12 in TM ＋ E2 ＋ G15 group increased（4.01 ± 0.46 vs 5.25 ± 0.80, 0.88 ±0.10 vs 1.02 ±0.07, P 〈0.05）, and the expressions of GRP78, caspase-12 and CHOP in TM ＋ E2 ＋ ICI ＋ G15 group increased （P 〈 0.05）. Compared with DTI＂ group, the expressions of GRP78 and CHOP in DTr ＋ E2group decreased （1.81 ± 0.08 vs 1.30± 0.14, 1.00 ± 0.13 vs 0.51 ± 0.01, P 〈 0.05 ）.

关 键 词：雌激素 内质网应激 人脐静脉血管内皮细胞 细胞凋亡 动脉粥样硬化 

分 类 号：R54[医药卫生—心血管疾病]