Background Cigarette smoking has an influence on both arterial-type and venous-type thrombosis. However, little is known about the direct effect of cigarette smoke extract （CSE） on fibrinolytic activity of human umb...Background Cigarette smoking has an influence on both arterial-type and venous-type thrombosis. However, little is known about the direct effect of cigarette smoke extract （CSE） on fibrinolytic activity of human umbilical vein endothelial cells （HUVECs）. Most recently, simvastatin has been marked in its effect on endothelial cells protection and anticoagulation. In this study, the effect of CSE on the expression of tissue-type plasminogen activator （t-PA） and plasminogen activator inhibitor-l（PAl-1） in HUVECs was addressed. The role of simvastatin in CSE-induced fibrinolytic activity changes was investigated as well. Methods The fourth to fifth generation of HUVECs were incubated respectively with 0, 5%, 10% and 20% CSE for 6 hours or exposed to 5% CSE for 0, 4, 6, 8, 12, 24 hours to determine the expression changes of t-PA and PAl-1 protein. Meanwhile, cells were also accordingly exposed either to 5% CSE alone or simvastatin pre-treated and 5% CSE for 24 hours to assess the role of simvastatin in CSE-induced t-PA and PAl-1 protein and mRNA expression in HUVECs. RT-PCR and ELISA techniques were used for detecting the t-PA or PAl-1 mRNA and protein. Results After 6-hour exposure to CSE, the expression levels of t-PA protein in 10% and 20% CSE-treated groups reduced significantly （（0.0365±0.0083） ng/ml, （0.0255±0.0087） ng/ml） when compared with that of control group （（0.0660±0.0120） ng/ml） （P 〈0.05）. In contrast, the levels of PAl-1 protein in 5%, 10% and 20% CSE-treated groups increased remarkably （（13.3225±0.5680） ng/ml, （14.2675±1.5380） ng/ml, （14.4292±1.6230） ng/ml） when compared with that of control group （（8.5193_±0.7537）ng/ml） （P〈0.05）. After stimulation with 5% CSE for 0, 4, 6, 8, 12, 24 hours, the levels of PAl-1 protein increased over time and reached the peak at 24 hours （（14.6400±1.0651） ng/ml）, which was significantly higher than that of control group （（12.0656±0.6148） ng/ml） （P 〈0.05）. Additionally, CSE could up-regulate PAl-1 expression at both the mRNA and the protein levels. The levels of PAl-1 mRNA and protein increased significantly in 5% CSE-treated group （（8.8030±0.4745） ng/ml, （1.8155±0.0412） ng/ml） compared with those of control groups （（5.0588±0.2315） ng/ml, （1.3030±0.0647） ng/ml） （P 〈0.01）, and decreased after 2-hour simvastatin pre-treatment （（5.4875±0.3166） ng/ml, （1.3975-±0.0297） ng/ml） （P 〈0.01）. No significant difference was found at the levels of t-PA protein and mRNA （P 〉0.05）. Conclusions CSE inhibits the fibrinolytic activity of HUVECs in vitro. Simvastatin plays a protective role in CSE-induced fibrinolytic malfunction.展开更多
A series of t-PA cDNA mutants containing different parts of 3’-UTR sequences have beenconstructed.In vitro translation of t-PA transcripts in rabbit reticulocyte lysates and its expression in COS-7cells show that th...A series of t-PA cDNA mutants containing different parts of 3’-UTR sequences have beenconstructed.In vitro translation of t-PA transcripts in rabbit reticulocyte lysates and its expression in COS-7cells show that the 3’-UTR sequence has a very strong inhibitory effect on t-PA translation.The deletion of3’-UTR results in 3—8-fold increase of t-PA expression.Further study shows that an AU-rich sequence of some200 nt at 3’ end of 3’-UTR is responsible for the translational inhibition.RNA stability experiment reveals thatthe AU-rich segment leads to a 3-fold decrease of t-PA mRNA stability.The insertion of this segment into the3’-UTR of luciferase gene results in an obvious inhibition of Luc expression.A model is proposed for theregulation of t-PA expression.展开更多
目的探讨血栓抽吸联合冠状动脉内注射重组人TNK组织型纤溶酶原激活剂对老年ST段抬高型心肌梗死患者急诊经皮冠状动脉介入治疗术中冠状动脉微循环及心功能的影响。方法回顾性选取2021年1月至2023年10月于天津市第三中心医院就诊的并行急...目的探讨血栓抽吸联合冠状动脉内注射重组人TNK组织型纤溶酶原激活剂对老年ST段抬高型心肌梗死患者急诊经皮冠状动脉介入治疗术中冠状动脉微循环及心功能的影响。方法回顾性选取2021年1月至2023年10月于天津市第三中心医院就诊的并行急诊经皮冠状动脉介入治疗的老年ST段抬高型心肌梗死患者90例,根据经皮冠状动脉介入治疗术中介入策略不同分为单纯血栓抽吸组(抽吸组)46例和血栓抽吸联合冠状动脉内注射重组人TNK组织型纤溶酶原激活剂组(联合组)44例。比较2组一般临床资料,术后90 min ST段回落指数≥70%比例,术后即刻心肌梗死溶栓试验(thrombolysis in myocardial infarction,TIMI)血流分级,术后TIMI心肌灌注分级,校正的TIMI血流帧数,心脏超声指标以及住院期间主要不良心血管事件及出血事件。结果联合组术后ST段回落≥70%、术后即刻TIMI血流分级3级、术后TIMI心肌灌注分级3级比例显著高于抽吸组,校正的TIMI血流帧数显著低于抽吸组(P<0.05);抽吸组术后1周的左心室射血分数显著低于联合组[(52.5±6.2)%vs(58.3±6.4)%,P<0.05],联合组术后1周左心室舒张末期内径显著低于抽吸组[(44.1±3.9)mm vs(51.9±2.5)mm,P<0.05];联合组住院期间主要不良心血管事件发生率显著低于抽吸组(20.5%vs 37.0%,P<0.05)。结论在应用抽吸导管的基础上配合冠状动脉内注射重组人TNK组织型纤溶酶原激活剂可有效降低老年ST段抬高型心肌梗死患者冠状动脉内血栓负荷,改善心肌微循环灌注,降低住院期间主要不良心血管事件发生率且不增加出血风险。展开更多
文摘Background Cigarette smoking has an influence on both arterial-type and venous-type thrombosis. However, little is known about the direct effect of cigarette smoke extract （CSE） on fibrinolytic activity of human umbilical vein endothelial cells （HUVECs）. Most recently, simvastatin has been marked in its effect on endothelial cells protection and anticoagulation. In this study, the effect of CSE on the expression of tissue-type plasminogen activator （t-PA） and plasminogen activator inhibitor-l（PAl-1） in HUVECs was addressed. The role of simvastatin in CSE-induced fibrinolytic activity changes was investigated as well. Methods The fourth to fifth generation of HUVECs were incubated respectively with 0, 5%, 10% and 20% CSE for 6 hours or exposed to 5% CSE for 0, 4, 6, 8, 12, 24 hours to determine the expression changes of t-PA and PAl-1 protein. Meanwhile, cells were also accordingly exposed either to 5% CSE alone or simvastatin pre-treated and 5% CSE for 24 hours to assess the role of simvastatin in CSE-induced t-PA and PAl-1 protein and mRNA expression in HUVECs. RT-PCR and ELISA techniques were used for detecting the t-PA or PAl-1 mRNA and protein. Results After 6-hour exposure to CSE, the expression levels of t-PA protein in 10% and 20% CSE-treated groups reduced significantly （（0.0365±0.0083） ng/ml, （0.0255±0.0087） ng/ml） when compared with that of control group （（0.0660±0.0120） ng/ml） （P 〈0.05）. In contrast, the levels of PAl-1 protein in 5%, 10% and 20% CSE-treated groups increased remarkably （（13.3225±0.5680） ng/ml, （14.2675±1.5380） ng/ml, （14.4292±1.6230） ng/ml） when compared with that of control group （（8.5193_±0.7537）ng/ml） （P〈0.05）. After stimulation with 5% CSE for 0, 4, 6, 8, 12, 24 hours, the levels of PAl-1 protein increased over time and reached the peak at 24 hours （（14.6400±1.0651） ng/ml）, which was significantly higher than that of control group （（12.0656±0.6148） ng/ml） （P 〈0.05）. Additionally, CSE could up-regulate PAl-1 expression at both the mRNA and the protein levels. The levels of PAl-1 mRNA and protein increased significantly in 5% CSE-treated group （（8.8030±0.4745） ng/ml, （1.8155±0.0412） ng/ml） compared with those of control groups （（5.0588±0.2315） ng/ml, （1.3030±0.0647） ng/ml） （P 〈0.01）, and decreased after 2-hour simvastatin pre-treatment （（5.4875±0.3166） ng/ml, （1.3975-±0.0297） ng/ml） （P 〈0.01）. No significant difference was found at the levels of t-PA protein and mRNA （P 〉0.05）. Conclusions CSE inhibits the fibrinolytic activity of HUVECs in vitro. Simvastatin plays a protective role in CSE-induced fibrinolytic malfunction.
文摘A series of t-PA cDNA mutants containing different parts of 3’-UTR sequences have beenconstructed.In vitro translation of t-PA transcripts in rabbit reticulocyte lysates and its expression in COS-7cells show that the 3’-UTR sequence has a very strong inhibitory effect on t-PA translation.The deletion of3’-UTR results in 3—8-fold increase of t-PA expression.Further study shows that an AU-rich sequence of some200 nt at 3’ end of 3’-UTR is responsible for the translational inhibition.RNA stability experiment reveals thatthe AU-rich segment leads to a 3-fold decrease of t-PA mRNA stability.The insertion of this segment into the3’-UTR of luciferase gene results in an obvious inhibition of Luc expression.A model is proposed for theregulation of t-PA expression.
文摘目的探讨血栓抽吸联合冠状动脉内注射重组人TNK组织型纤溶酶原激活剂对老年ST段抬高型心肌梗死患者急诊经皮冠状动脉介入治疗术中冠状动脉微循环及心功能的影响。方法回顾性选取2021年1月至2023年10月于天津市第三中心医院就诊的并行急诊经皮冠状动脉介入治疗的老年ST段抬高型心肌梗死患者90例,根据经皮冠状动脉介入治疗术中介入策略不同分为单纯血栓抽吸组(抽吸组)46例和血栓抽吸联合冠状动脉内注射重组人TNK组织型纤溶酶原激活剂组(联合组)44例。比较2组一般临床资料,术后90 min ST段回落指数≥70%比例,术后即刻心肌梗死溶栓试验(thrombolysis in myocardial infarction,TIMI)血流分级,术后TIMI心肌灌注分级,校正的TIMI血流帧数,心脏超声指标以及住院期间主要不良心血管事件及出血事件。结果联合组术后ST段回落≥70%、术后即刻TIMI血流分级3级、术后TIMI心肌灌注分级3级比例显著高于抽吸组,校正的TIMI血流帧数显著低于抽吸组(P<0.05);抽吸组术后1周的左心室射血分数显著低于联合组[(52.5±6.2)%vs(58.3±6.4)%,P<0.05],联合组术后1周左心室舒张末期内径显著低于抽吸组[(44.1±3.9)mm vs(51.9±2.5)mm,P<0.05];联合组住院期间主要不良心血管事件发生率显著低于抽吸组(20.5%vs 37.0%,P<0.05)。结论在应用抽吸导管的基础上配合冠状动脉内注射重组人TNK组织型纤溶酶原激活剂可有效降低老年ST段抬高型心肌梗死患者冠状动脉内血栓负荷,改善心肌微循环灌注,降低住院期间主要不良心血管事件发生率且不增加出血风险。
