可溶性endoglin对人脐静脉内皮细胞产生一氧化氮及其合成酶磷酸化水平的影响  

Effects of soluble endoglin on nitric oxide production and nitric oxide synthase phosphorylation in cultured human umbilical vein endothelial cells

作　　者：董微[1] 许群星[1] 韩玉环[1] 尹利荣[1] 张立军[1] 华绍芳[1] 

机构地区：[1]天津医科大学第二医院产科,300211

出　　处：《中华围产医学杂志》2011年第11期682-687,共6页Chinese Journal of Perinatal Medicine

基　　金：天津市自然科学基金（09JCYBJC13000）

摘　　要：目的观察可溶性endoglin（soluble endoglin，sEng）对体外培养的人脐静脉内皮细胞产生一氧化氮（nitric oxide，NO）、内皮型一氧化氮合酶（endothelial nitric oxide synthase，eNOS）及其1177位点丝氨酸磷酸化程度的影响。方法将3代以内的人脐静脉内皮细胞接种于96孔培养板，分别加入完全细胞培养液（对照组）和不同浓度的sEng（1、10、100μg／L），作用6、12和24h后收取细胞培养液及细胞，硝酸酶还原法测定细胞培养液中NO代谢产物亚硝酸盐浓度反映NO含量，Western印记法检测各组细胞eNOS的相对表达量及其1177位点丝氨酸磷酸化情况，实时荧光聚合酶链反应技术检测各组细胞eNOSmRNA的相对表达量。采用方差分析、LSD法及Pearson相关分析法对各组进行比较。结果（1）1、10、100μg／LsEng作用6h，细胞培养液中NO浓度分别为（59．25±1．63）、（41．08±2．71）和（30．38±1．63）μmol／L；作用12h，细胞培养液中NO浓度分别为（54．98±3．34）、（35．00±8．60）和（19．82±3．75）μmol／L；作用24h，细胞培养液中NO浓度分别为（46．14±4．93）、（30．24±2．08）和（12．78±5．01）μml／L，而对照组NO浓度随着时间的推移无明显改变（F=2．30，P=0．14）。与sEng共培养后，细胞培养液中NO浓度明显降低，并与sEng作用时间（r=-0．98，P〈0．05）及浓度（=-0．88，P〈0．05）呈明显的负相关。（2）1、10、100μg／LsEng作用6heNOS相对表达量分别为0．71±0．00、0．47±0．00和0．32±0．00；作用12h，eNOS相对表达量分别为0．58±0．00、0．42±0．00和0．25±0．00；作用24h，eNOS相对表达量分别为0．49±0．00、0．33±0．00和0．18±0．00。而对照组eNOS相对表达量及1177位点丝氨酸磷酸化eNOS吸光度／总eNOS吸光度随时间推移无明显改变（F分别为3．59和0．37，P分别为0．09和0．80）。与sEng共培养后，细胞中eNOS蛋白表�Objective To investigate the effects of soluble endoglin （sEng） on nitric oxide （NO） production and endothelial nitric oxide synthase （eNOS） phosphorylation in cultured human umbilical vein endothelial cells. Methods Human umbilical vein endothelial cells within 3 passages seeded in culture plates of 96 wells, were stimulated by total culture medium （control group） or sEng （1, 10 and 100 g/L） respectively. Cells and medium were collected after cells were cultured for 6, 12 and 24 hours respectively. The concentration of the metabolites of NO in each group was measured by nitrate reductase method. The expression of eNOS and eNOS-Set（p）1177 were detected by Western blot. The expression of eNOS mRNA in each group was detected by real-time fluorescence reverse transcription-polymerase chain reaction. Analysis of variance, LSD method and pearson correlation were used to compare the difference between groups. Results （1）The concentration of the metabolites of NO in 1, 10 and 100μg/L sEng groups was （59.25±1.63）, （41.08±2.71） and （30.38±1.63） μmol/L respectively after cultured for 6 hours; （54.98±3.34）, （35.00±8.60） and （19.82±3.75） μmol/L for 12 hours; and （46.14±4.93）, （30.24±2.08） and （12.78±5.01） μmol/L for 24 hours. There was no significant changes in control group with time going by （F=2.30, P= 0.14）. The concentration of the metabolites of NO was significantly lower in sEng group, and which had negative correlation with culture time （r= -0.98, P〈0.05） and dose （r= -0.88, P〈0.05）. （2） The expression of eNOS in 1, 10, 100 μg/L sEng groups was 0.71±0.00, 0.47 ± 0.00 and 0.32!0.00 after cultured for 6 hours; 0.58±0.00, 0.42±0.00 and 0.25±0.00 for 12 hours; and 0.49±0.00, 0.33±0.00 and 0.18±0. 00 for 24 hours. While the expression of eNOS and eNOS-Ser （p）1177/eNOS had no significant changes in control group with time going by （F=3.59 and 0.37, P 0.09 and 0.80）. The expression of eNOS protein and eNOS-Ser�

关 键 词：受体 细胞表面 抗原 CD 脐静脉 内皮细胞 一氧化氮 一氧化氮合酶 

分 类 号：R363[医药卫生—病理学]