人脐静脉内皮细胞分泌组织型纤溶酶原激活物及其抑制剂1对醒脑静干预的反应  

Effects of Xingnaojing on recombinant human tumor necrosis factor-mediated tissue-type plasminogen activator and plasminogen activator inhibitor-1 expression in human umbilical cord vein endothelial cells

作　　者：欧阳海春 吴沃栋[2] 钟冬梅[3] 吴焱贤 李文杰[2] 陈晞明[2] 

机构地区：[1]广东省佛山市顺德区第一人民医院心血管内科,广东省佛山市528300 [2]广州医科大学附属第三医院心血管内科,广东省广州市510150 [3]广州医科大学附属第三医院中医科,广东省广州市510150

出　　处：《中国组织工程研究》2015年第11期1717-1721,共5页Chinese Journal of Tissue Engineering Research

基　　金：广州市中医药局中西医结合科研课题(2004A014);广东省中医药局建设中医药强省科研课题(2060063)~~

摘　　要：背景:前期研究发现醒脑静能有效抑制兔心肌缺血再灌注模型的炎症递质及促纤溶作用,但是影响纤溶系统活性的作用机制未完全明确。目的:观察醒脑静对重组人肿瘤坏死因子α介导人脐静脉内皮细胞分泌组织型纤溶酶原激活物和纤溶酶原激活物抑制剂1及其基因表达的影响。方法:取3-5代人脐静脉内皮细胞,在培养基中添加10μg/L重组人肿瘤坏死因子α介导人脐静脉内皮细胞分泌组织型纤溶酶原激活物和纤溶酶原激活物抑制剂1及其基因表达,醒脑静组加入不同浓度(5,10,20 m L/L)的醒脑静干预,阳性对照组添加氟伐他汀(1μmol/L),并设立单纯人脐静脉内皮细胞培养的空白对照组。培育24 h后采用酶联免疫吸附双抗体夹心法(ELISA)检测细胞上清液组织型纤溶酶原激活物和纤溶酶原激活物抑制剂1水平;采用反转录聚合酶链反应检测人脐静脉内皮细胞的组织型纤溶酶原激活物和纤溶酶原激活物抑制剂1的m RNA表达。结果与结论:重组人肿瘤坏死因子α组纤溶酶原激活物抑制剂1分泌和m RNA表达较空白对照组显著升高(P<0.05),组织型纤溶酶原激活物分泌和m RNA表达较空白对照组显著降低(P<0.05)。不同浓度醒脑静组纤溶酶原激活物抑制剂1分泌和m RNA表达均较重组人肿瘤坏死因子α组显著降低(P<0.05),而组织型纤溶酶原激活物分泌和m RNA表达均较重组人肿瘤坏死因子α组显著升高(P<0.05),且呈剂量依赖关系。结果证实,醒脑静作用可逆转重组人肿瘤坏死因子α所致的人脐静脉内皮细胞的纤溶活性。BACKGROUND：We have found thatXingnaojing can effectively inhibit mediators of inflammatory mediators and promote fibrinolytic activity in a rabbit model of myocardial ischemia and reperfusion. However, it is not completely clear what is the action of mechanism of fibrinolytic system activity in human umbilical vein endothelial cels. OBJECTIVE：To investigate the effects ofXingnaojingon recombinant human tumor necrosis factor α-mediated tissue-type plasminogen activator （t-PA） and plasminogen activator inhibitor-1 （PAI-1） expression in human umbilical cord vein endothelial cels. METHODS： The passages 3-5 of human umbilical cord vein endothelial cels were obtained, and were induced by adding 10 μg/L recombinant human tumor necrosis factor α in the culture medium.Xingnaojing in different concentrations （5, 10, 20 mL/L） were injected in theXingnaojing group, and fluvastatin （1 μmol/L） was added as positive control group. A blank control group of simple human umbilical cord vein endothelial cels culture was set up. After 24 hours of cultivation, the production of t-PA and PAI-1 in the cultured medium was determined by ELISA. The mRNA expression of t-PA and PAI-1 was measured by reverse transcript-PCR. RESULTS AND CONCLUSION：Compared with the blank control group, the secretion and mRNA expression of PAI-1 in the group treated with recombinant human tumor necrosis factor α（10μg/L） were significantly increased （P 〈 0.05）; however, the secretion and mRNA expression of t-PA in the group treated with recombinant human tumor necrosis factorα （10μg/L） were significantly decreased （P 〈 0.05）. The secretion and mRNA expression of t-PA were significantly higher in theXingnaojinggroup at different concentrations than in the recombinant human tumor necrosis factor α group （P 〈 0.05）; however, the secretion and mRNA expression of PAI-1 were significantly lower in the former group than the latter group （P〈 0.05）, which was in a dose-dependent manner.Xingnaojing can ef

关 键 词：组织构建 血管内皮细胞 醒脑静 安宫牛黄丸 肿瘤坏死因子 内皮细胞 组织型纤溶酶原激活物 纤溶酶原激活物抑制剂1 

分 类 号：R318[医药卫生—生物医学工程]