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作 者:陈爱华[1] 肖华[1] 李志梁[1] 吴金家[1] 季爱民[1]
出 处:《中草药》2006年第7期1045-1048,共4页Chinese Traditional and Herbal Drugs
基 金:广东省科技计划项目(粤科计字[2004]115号)
摘 要:目的 观察前荷叶碱对血管紧张素Ⅱ(AngⅡ)诱导人脐静脉内皮细胞(HUVECs)凋亡的影响,探讨其对HUVECs的保护作用机制。方法 体外培养HUVECs细胞系ECV304,以10μmol/L卡托普利或10、1、0.1、0.01μmol/L前荷叶碱作用于HUVECs,30min后再加入AngⅡ1μmol/L,光镜下观察细胞形态,MTT法观察前荷叶碱对HUVECs活性的影响,比色法测定NO、总一氧化氮合酶(tNOS)、诱导型一氧化氮合酶(iNOS)水平,流式细胞仪测定细胞凋亡率。结果 与对照组比较,AngⅡ能明显诱导HUVECs的凋亡(P〈0.01),10μmol/L卡托普利及0.01~10μmol/L前荷叶碱可明显改善内皮细胞形态,组织活性明显升高(P〈0.05),能增加HUVECs释放NO及生成tNOS(P〈0.05),但对iNOS影响不大,使AngⅡ诱导的HUVECs凋亡细胞数明显减少(P〈0.05)。结论 前荷叶碱通过增加NO生成而抑制AngⅡ诱导的HUVECs凋亡,从而发挥可能的内皮保护功能。Objective To investigate the effect of pronuciferine on apoptosis of cultured human umbilical vein endothelium cells (HUVECs) induced by angiotensin Ⅱ(Ang Ⅱ ). Methods HUVECs cell line ECV304 was cultured in vitro, pretreated with Captopril (10μmol/L) or pronuciferine 10, 1, 0. 1, 0. 01μmol/L for 30min, respectively, then treated with AngⅡ (1μmol/L). Cell-morphosis was observed by light microscope. Cells viability was assessed by MTT assay. Production of nitric oxide (NO), activities of total nitric oxide synthase (tNOS), and inducible nitric oxide synthase (iNOS) were measured by colorimetry. Apoptosis rate was measured by Flow Cytometer (FCM). Results Ang Ⅱ induced typical endothelial cell apoptosis and the apoptosis rates were significantly higher than those of the control group (P〈0.01). The cell-morphosis was improved by Captopril and pronuciferine. Captopril and pronuciferine could significantly increased the production of NO and the activity of tNOS, but pronuciferine had no effect on activity of iNOS. Captopril and pronuciferine significantly inhibited the apoptosis rate of ECV304 cells induced by AngⅡ with reducing the apoptotic cell number. Conclusion Pronuciferine could decrease the apoptosis of ECV304 cells induced by AngⅡ through increasing the level of NO so that it is potential to protect endothelium function.
关 键 词:前荷叶碱 人脐静脉内皮细胞(HUVECs) 血管紧张素Ⅱ(AngⅡ) 细胞凋亡
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