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正五聚蛋白3对血管紧张素Ⅱ刺激后血管内皮细胞凋亡及血管活性物质分泌的影响 预览

Effect of PTX3 on apoptosis and vasoactive substance secretion of vascular endothelial cells stimulated by Ang Ⅱ
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摘要 目的探讨正五聚蛋白(PTX)3对血管紧张素(Ang)Ⅱ刺激后的血管内皮细胞(HUVECs)凋亡及血管活性物质分泌的影响。方法以10-7mol/LAngⅡ刺激体外培养的HUVECs0、12、24和48h后,Western印迹检测HUVECs中PTX3蛋白的表达。利用Lipofectamine2000将PTX3siRNA转染至HUVECs,Western印迹检测其转染效果。将HUVECs随机分为空白组(未做任何处理)、AngⅡ组(10-7mol/LAngⅡ刺激)、AngⅡ+siNC组(转染阴性对照序列后,给予10-7mol/LAngⅡ刺激)和AngⅡ+siPTX3组(转染PTX3siRNA后,给予10-7mol/LAngⅡ刺激),噻唑蓝(MTT)法检测各组细胞的增殖情况,流式细胞仪检测各组细胞的凋亡,实时聚合酶链反应(RT-PCR)检测各组细胞中血管活性物质内皮素(ET)-1和诱导型一氧化氮合酶(iNOS)mRNA的表达。结果随着AngⅡ刺激时间的延长,PTX3蛋白表达水平显著升高。转染PTX3siRNA后成功下调了AngⅡ引起的PTX3表达。AngⅡ刺激可引起HUVECs增殖减弱,凋亡增强,ET-1和iNOSmRNA表达升高;下调PTX3表达能够减弱AngⅡ引起的HUVECs损伤,促进HUVECs增殖,抑制凋亡,并下调ET-1和iNOSmRNA表达。结论PTX3在AngⅡ刺激后HUVECs中表达升高,下调其表达可抑制AngⅡ诱导的细胞凋亡及血管活性物质的分泌。 Objective To investigate the effects of pentamer (PTX)3 on apoptosis and secretion of vasoactive substances in human umbilical vascular endothelial cells (HUVECs) stimulated by AngⅡ. Methods After stimulating HUVECs with 10 -7 mol/L AngⅡ for 0, 12, 24 and 48 hours in vitro, the expression of PTX3 protein in HUVECs was detected by Western blot. PTX3 siRNA was transfected into HUVECs by Lipofectamine 2000, and its transfection effect was detected by Western blot. HUVECs were randomly divided into blank group (without any treatment), AngⅡ group (10 -7 mol/L AngⅡ stimulation), AngⅡ+ siNC group (10 -7 mol/L AngⅡ stimulation after transfection of negative control sequence) and AngⅡ+ siPTX3 group 10 -7 mol/L AngⅡ stimulation after transfection of PTX3 siRNA). Cell proliferation was detected by MTT method, cell apoptosis was detected by flow cytometry, and the expression of endothelin (ET)-1 and inducible nitric oxide synthase (iNOS) mRNA in cells of each group were detected by RT-PCR. Results The expression of PTX3 protein was increased significantly with the prolongation of AngⅡ stimulation time. PTX3 siRNA was successfully transfected to down-regulate PTX3 expression induced by AngⅡ. AngⅡ stimulation induced HUVECs to decrease proliferation, increase apoptosis, and increase the expressions of ET-1 and iNOS. Down-regulation of PTX3 expression attenuated HUVECs injury induced by AngⅡ, promoted HUVECs proliferation, inhibited apoptosis, and down-regulated the expressions of ET-1 and iNOS. Conclusions The expression of PTX3 is increased in HUVECs stimulated by AngⅡ, and its down regulation can inhibit the apoptosis induced by AngⅡ and the secretion of vasoactive substances.
作者 杨雁华 汤建民 来桂棵 王丰云 YANG Yan-Hua;TANG Jian-Min;LAI Gui-Ke(Department of Cardiology, the Second Affiliated Hospital of Zhengzhou University, Zhengzhou 450000, Henan, China)
出处 《中国老年学杂志》 CAS 北大核心 2019年第9期2193-2197,共5页 Chinese Journal of Gerontology
基金 河南省科技发展计划项目(182102310521).
关键词 血管内皮细胞 血管紧张素Ⅱ 正五聚蛋白(PTX)3 细胞凋亡 内皮素-1 诱导型一氧化氮合酶 Vascular endothelial cells Ang Ⅱ PTX3 Apoptosis ET-1 iNOS
作者简介 第一作者:杨雁华(1984-),女,硕士,主治医师,主要从事冠心病、心力衰竭研究。
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