尼古丁对内皮细胞纤溶损害的机制及辛伐他汀的干预作用
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摘要
研究背景
吸烟是心血管疾病发生发展的一个独立而重要的危险因素。吸烟可以损伤血管内皮细胞,导致内皮细胞功能障碍,引起体内纤溶系统的异常变化,从而增加血栓形成的风险。众所周知,血液的纤溶能力主要取决于组织型纤溶酶原激活剂(t-PA)及其快速抑制剂纤溶酶原激活物抑制物-1(PAI-1)的活性。大量文献表明,糖尿病、冠心病可伴发纤溶功能异常,而有关吸烟引起的纤溶变化研究尚少。尼古丁是烟草的主要致病成分之一,其通过损伤内皮细胞导致血液纤溶功能下降,但该作用的具体机制尚不十分清楚。蛋白激酶C(PKC)为细胞内信号转导的重要递质,广泛参与细胞的分泌、增殖、分化等功能,其作用日益受到关注。目前,尼古丁所致内皮细胞纤溶功能下降与PKC通道的关系研究较少,且结论不一。辛伐他汀为临床常用的血脂调节剂,研究表明该药还具有内皮保护、抗炎、促进纤溶等作用。辛伐他汀是否对尼古丁介导的人脐静脉内皮细胞纤溶功能紊乱具有保护作用及其可能机制目前尚无报道。
目的
研究尼古丁及不同浓度的PKC阻断剂Staurosporine(STS)作用于人脐静脉内皮细胞(HUVECs)后其t-PA、PAI-1蛋白及mRNA表达的变化,探讨尼古丁对HUVECs纤溶影响的可能机制;采用不同浓度的辛伐他汀预处理内皮细胞,了解该药对尼古丁介导的HUVECs表达t-PA、PAI-1蛋白及mRNA的影响。
方法
1.实验分组HUVECs株培养后接种于25cm×25cm的培养瓶中,随机分为①对照组:培养液中加入与尼古丁相同体积的无菌PBS液;②尼古丁组:培养液中加入终浓度为100μmol/L的尼古丁;③STS干预组:培养液中分别加入终浓度为20、50、100μmol/L的STS预处理细胞30min,再加入100μmol/L的尼古丁共同培养;④辛伐他汀干预组:培养液中分别加入终浓度为1、10、100μmol/L的辛伐他汀预处理细胞2h,再加入100μmol/L的尼古丁共同培养。
2.检测方法同步培养24h后收集各组的细胞及上清液,采用酶联免疫吸附双抗体夹心法(ELISA)测定各组细胞上清液中t-PA、PAI-1蛋白浓度,逆转录-聚合酶链反应法(RT-PCR)检测各组细胞中t-PA和PAI-1 mRNA的表达水平。
结果
1.尼古丁组PAI-1mRNA及蛋白含量〔0.969±0.112,(66.051±2.890)ng/ml〕与对照组〔0.099±0.013,(38.660±1.749)ng/ml〕比较显著升高,差异均有统计学意义(均P﹤0.01)。尼古丁组t-PA mRNA的表达(0.099±0.011)与对照组(1.011±0.132)比较明显下降,差异有统计学意义(P﹤0.01);尼古丁组t-PA蛋白(0.725±0.211)ng/ml表达与对照组(1.082±0.131)ng/ml比较,差异无统计学意义(P>0.05)。
2.STS干预结果各浓度STS干预组PAI-1mRNA及蛋白〔0.588±0.077,(61.922±5.821)ng/ml〕、〔0.530±0.070,(57.976±8.009)ng/ml〕、〔0.222±0.029,(43.291±1.878)ng/ml〕均较尼古丁组降低,差异均有统计学意义(均P﹤0.01);各STS干预组PAI-1mRNA及蛋白高于对照组,差异均有统计学意义(均P﹤0.01)。20μmol/LSTS组t-PA mRNA(0.184±0.023)与尼古丁组比较,差异无统计学意义(P>0.05),50、100μmol /L STS组t-PA mRNA(0.293±0.037,0.412±0.053)较尼古丁组升高,但仍低于对照组,差异均有统计学意义(均P﹤0.01);各组t-PA蛋白与对照组及尼古丁组比较,差异均无统计学意义(均P>0.05)。
3.辛伐他汀干预结果各浓度辛伐他汀干预组PAI-1mRNA及蛋白〔0.581±0.095,(60.788±2.500)ng/ml〕、〔0.511±0.084,(54.955±3.971)ng/ml〕、〔0.0219±0.037,(41.924±4.472)ng/ml〕均较尼古丁组降低,差异均有统计学意义(均P﹤0.01),以100μmol/L辛伐他汀组降低最为显著,与对照组比较差异无统计学意义(P>0.05)。1μmol /L辛伐他汀组t-PA mRNA(0.187±0.025)与尼古丁组比较,差异无统计学意义(P>0.05);10、100μmol/L辛伐他汀组t-PA mRNA(0.278±0.038)、(0.332±0.046)较尼古丁组升高,但仍低于对照组,差异均有统计学意义(均P﹤0.01);各组t-PA蛋白与对照组及尼古丁组比较,差异均无统计学意义(均P>0.05)。
结论
1.尼古丁通过上调HUVECs PAI-1蛋白和mRNA的表达,对内皮细胞的纤溶活性具有抑制作用。
2.尼古丁介导的内皮细胞纤溶紊乱部分通过P KC通道起作用。
3.辛伐他汀对尼古丁所致的内皮细胞纤溶损害具有保护作用。
Backgroud
Cigarette smoking was considered as a dependent danger factor to the development of cardio- vascular disease .It could damage the vascular endothelial cells, lead to endothelial cells dysfunction, as well as trigger fibrinolysis imbalance, then increasing the risk of thrombosis. As we all know, fibrinolytic system relies heavily on the activity of tissue-type plasminogen activator ( t-PA )and tissue-type plasminogen inhibitor-1(PAI-1) released by vascular endothelial cells. There has many reports about the disfunction of fibrinolysis studies associated with diabetes , coronary heart disease .But studies about disfunction of fibrinolysis associated with cigarette smoking was reported few. Nicotine, one of the main harmful substances of smoking, can damage the fibrinolytic function of endothelial cells, and its related mechanism was entirely unclear .Protein kinase C (PKC) is an important biological intracellular signal transduction pathway in the body, it widespread participate the secretion ,proliferation and differentiation of cells, its get to pay close attention increasingly .Now, the studies about nicotine decrease the fibrinolytic of endothelial cells associate with PKC is few reported. Simvastatin as a represent of liqid- lowering statin drug, it has many new function ,for example ,the protection of endothelial cells, inhibit inflammation, promote fibrolysis ,etc. Simvastatin can wether or not improved fibrinolytic activity induced by nicotine and its related mechanism has not been reported.
Objective
To study the effect of nicotine and different concentrations of staurosporine on the expression of t-PA and PAI-1 protein and mRNA in human umbilical vein endothelial cell (HUVECs), To explore the mechanism of fibrinolytic function stimulated by nicotine . Using different concentration of simvastatin to pretreatment the HUVECs, and observed the influence of simvastatin to the expression of t-PA,PAI-1 protein and mRNA stimulated by nicotine.
Methods
Experimental Group:HUVECs were cultured in 25cm2 culture flasks and randomly divided into①the control group②100μmol/L nicotine group③STS intervention group. The cells were treated with STS at concentration of 20、50、100μmol/L for 30min,and then exposed to nicotine at concentration of 100μmol/L.④Simvastatin intervention group .The cells were treated with simvastatin at concentration of 1、10、100μmol/L for 2h,and then exposed to nicotine at concentration of 100μmol/L.The cells and supernatants of each group were collected after 24 hours cultured .The expression of t-PA and PAI-1 protein were measured by ELISA and the RNA expression of t-PA and PAI-1 was determined by RT-PCR.
Results
1.Compared with control group〔0.099±0.013(,38.660±1.749)ng/ml〕, the PAI-1 mRNA and protein expression of nicotine group〔0.969±0.112,(66.051±2.890)ng/ml〕were increased significantly(P<0.01, respectively).Compared with control group(1.011±0.132),the t-PA mRNA(0.099±0.011)expression of nicotine group were decreased significantly(P<0.01).However, the expression of t-PA protein was no significantly different(P>0.05).
2.Compared with nicotine group , the PAI-1 mRNA and protein expression of different concentrations of STS intervention group〔0.588±0.077,(61.922±5.821)ng/ml〕、〔0.530±0.070,(57.976±8.009)ng/ml〕、〔0.222±0.029,(43.291±1.878)ng/ml〕were decreased significantly ,but still higher than control group(P<0.01, respectively). Compared with nicotine group, the t-PA mRNA expression of 20μmol/L STS group(0.184±0.023)were increased not significantly(P>0.05);The t-PA mRNA expression of 50、100μmol/L STS group(0.293±0.037)、(0.412±0.053)were increased significantly ,but still lower than control group(P<0.01, respectively).The expression of t-PA protein was no significantly different (P>0.05).
3.Compared with nicotine group , the PAI-1 mRNA and protein expression of different concentrations of simvastatin intervention group〔0.581±0.095,(60.788±2.500)ng/ml〕、〔0.511±0.084,(54.955±3.971)ng/ml〕、〔1.074±0.099,(16.190±2.149)ng/ml〕were decreased significantly ,and the most notably effect of simvastatin was at the concentrations of 100μmol/L ,almost the same as that in the control group(P>0.05). Compared with nicotine group, the t-PA mRNA expression of 1μmol/L simvastatin group(0.187±0.025)were increased not significant(P>0.05);The t-PA mRNA expression of 10、100μmol/L simvastatin group(0.278±0.038)、(0.332±0.046)were increased significantly ,but still lower than control group(P<0.01, respectively). The expression of t-PA protein was no significantly different (P>0.05).
Conclusions
1.Nicotine through increased the expression of PAI-1 protein and mRNA in HUVECs,inhibits the fibrinolytic function of endothelial cells.
2.Activation the PKC can partly disorder the fibrinolytic function of endothelial cells that induced by nicotine.
3.Simvastatin can improves the fibrinolytic function of endothelial cells that induced by nicotine
吸烟是心血管疾病发生发展的一个独立而重要的危险因素。吸烟可以损伤血管内皮细胞,导致内皮细胞功能障碍,引起体内纤溶系统的异常变化,从而增加血栓形成的风险。众所周知,血液的纤溶能力主要取决于组织型纤溶酶原激活剂(t-PA)及其快速抑制剂纤溶酶原激活物抑制物-1(PAI-1)的活性。大量文献表明,糖尿病、冠心病可伴发纤溶功能异常,而有关吸烟引起的纤溶变化研究尚少。尼古丁是烟草的主要致病成分之一,其通过损伤内皮细胞导致血液纤溶功能下降,但该作用的具体机制尚不十分清楚。蛋白激酶C(PKC)为细胞内信号转导的重要递质,广泛参与细胞的分泌、增殖、分化等功能,其作用日益受到关注。目前,尼古丁所致内皮细胞纤溶功能下降与PKC通道的关系研究较少,且结论不一。辛伐他汀为临床常用的血脂调节剂,研究表明该药还具有内皮保护、抗炎、促进纤溶等作用。辛伐他汀是否对尼古丁介导的人脐静脉内皮细胞纤溶功能紊乱具有保护作用及其可能机制目前尚无报道。
目的
研究尼古丁及不同浓度的PKC阻断剂Staurosporine(STS)作用于人脐静脉内皮细胞(HUVECs)后其t-PA、PAI-1蛋白及mRNA表达的变化,探讨尼古丁对HUVECs纤溶影响的可能机制;采用不同浓度的辛伐他汀预处理内皮细胞,了解该药对尼古丁介导的HUVECs表达t-PA、PAI-1蛋白及mRNA的影响。
方法
1.实验分组HUVECs株培养后接种于25cm×25cm的培养瓶中,随机分为①对照组:培养液中加入与尼古丁相同体积的无菌PBS液;②尼古丁组:培养液中加入终浓度为100μmol/L的尼古丁;③STS干预组:培养液中分别加入终浓度为20、50、100μmol/L的STS预处理细胞30min,再加入100μmol/L的尼古丁共同培养;④辛伐他汀干预组:培养液中分别加入终浓度为1、10、100μmol/L的辛伐他汀预处理细胞2h,再加入100μmol/L的尼古丁共同培养。
2.检测方法同步培养24h后收集各组的细胞及上清液,采用酶联免疫吸附双抗体夹心法(ELISA)测定各组细胞上清液中t-PA、PAI-1蛋白浓度,逆转录-聚合酶链反应法(RT-PCR)检测各组细胞中t-PA和PAI-1 mRNA的表达水平。
结果
1.尼古丁组PAI-1mRNA及蛋白含量〔0.969±0.112,(66.051±2.890)ng/ml〕与对照组〔0.099±0.013,(38.660±1.749)ng/ml〕比较显著升高,差异均有统计学意义(均P﹤0.01)。尼古丁组t-PA mRNA的表达(0.099±0.011)与对照组(1.011±0.132)比较明显下降,差异有统计学意义(P﹤0.01);尼古丁组t-PA蛋白(0.725±0.211)ng/ml表达与对照组(1.082±0.131)ng/ml比较,差异无统计学意义(P>0.05)。
2.STS干预结果各浓度STS干预组PAI-1mRNA及蛋白〔0.588±0.077,(61.922±5.821)ng/ml〕、〔0.530±0.070,(57.976±8.009)ng/ml〕、〔0.222±0.029,(43.291±1.878)ng/ml〕均较尼古丁组降低,差异均有统计学意义(均P﹤0.01);各STS干预组PAI-1mRNA及蛋白高于对照组,差异均有统计学意义(均P﹤0.01)。20μmol/LSTS组t-PA mRNA(0.184±0.023)与尼古丁组比较,差异无统计学意义(P>0.05),50、100μmol /L STS组t-PA mRNA(0.293±0.037,0.412±0.053)较尼古丁组升高,但仍低于对照组,差异均有统计学意义(均P﹤0.01);各组t-PA蛋白与对照组及尼古丁组比较,差异均无统计学意义(均P>0.05)。
3.辛伐他汀干预结果各浓度辛伐他汀干预组PAI-1mRNA及蛋白〔0.581±0.095,(60.788±2.500)ng/ml〕、〔0.511±0.084,(54.955±3.971)ng/ml〕、〔0.0219±0.037,(41.924±4.472)ng/ml〕均较尼古丁组降低,差异均有统计学意义(均P﹤0.01),以100μmol/L辛伐他汀组降低最为显著,与对照组比较差异无统计学意义(P>0.05)。1μmol /L辛伐他汀组t-PA mRNA(0.187±0.025)与尼古丁组比较,差异无统计学意义(P>0.05);10、100μmol/L辛伐他汀组t-PA mRNA(0.278±0.038)、(0.332±0.046)较尼古丁组升高,但仍低于对照组,差异均有统计学意义(均P﹤0.01);各组t-PA蛋白与对照组及尼古丁组比较,差异均无统计学意义(均P>0.05)。
结论
1.尼古丁通过上调HUVECs PAI-1蛋白和mRNA的表达,对内皮细胞的纤溶活性具有抑制作用。
2.尼古丁介导的内皮细胞纤溶紊乱部分通过P KC通道起作用。
3.辛伐他汀对尼古丁所致的内皮细胞纤溶损害具有保护作用。
Backgroud
Cigarette smoking was considered as a dependent danger factor to the development of cardio- vascular disease .It could damage the vascular endothelial cells, lead to endothelial cells dysfunction, as well as trigger fibrinolysis imbalance, then increasing the risk of thrombosis. As we all know, fibrinolytic system relies heavily on the activity of tissue-type plasminogen activator ( t-PA )and tissue-type plasminogen inhibitor-1(PAI-1) released by vascular endothelial cells. There has many reports about the disfunction of fibrinolysis studies associated with diabetes , coronary heart disease .But studies about disfunction of fibrinolysis associated with cigarette smoking was reported few. Nicotine, one of the main harmful substances of smoking, can damage the fibrinolytic function of endothelial cells, and its related mechanism was entirely unclear .Protein kinase C (PKC) is an important biological intracellular signal transduction pathway in the body, it widespread participate the secretion ,proliferation and differentiation of cells, its get to pay close attention increasingly .Now, the studies about nicotine decrease the fibrinolytic of endothelial cells associate with PKC is few reported. Simvastatin as a represent of liqid- lowering statin drug, it has many new function ,for example ,the protection of endothelial cells, inhibit inflammation, promote fibrolysis ,etc. Simvastatin can wether or not improved fibrinolytic activity induced by nicotine and its related mechanism has not been reported.
Objective
To study the effect of nicotine and different concentrations of staurosporine on the expression of t-PA and PAI-1 protein and mRNA in human umbilical vein endothelial cell (HUVECs), To explore the mechanism of fibrinolytic function stimulated by nicotine . Using different concentration of simvastatin to pretreatment the HUVECs, and observed the influence of simvastatin to the expression of t-PA,PAI-1 protein and mRNA stimulated by nicotine.
Methods
Experimental Group:HUVECs were cultured in 25cm2 culture flasks and randomly divided into①the control group②100μmol/L nicotine group③STS intervention group. The cells were treated with STS at concentration of 20、50、100μmol/L for 30min,and then exposed to nicotine at concentration of 100μmol/L.④Simvastatin intervention group .The cells were treated with simvastatin at concentration of 1、10、100μmol/L for 2h,and then exposed to nicotine at concentration of 100μmol/L.The cells and supernatants of each group were collected after 24 hours cultured .The expression of t-PA and PAI-1 protein were measured by ELISA and the RNA expression of t-PA and PAI-1 was determined by RT-PCR.
Results
1.Compared with control group〔0.099±0.013(,38.660±1.749)ng/ml〕, the PAI-1 mRNA and protein expression of nicotine group〔0.969±0.112,(66.051±2.890)ng/ml〕were increased significantly(P<0.01, respectively).Compared with control group(1.011±0.132),the t-PA mRNA(0.099±0.011)expression of nicotine group were decreased significantly(P<0.01).However, the expression of t-PA protein was no significantly different(P>0.05).
2.Compared with nicotine group , the PAI-1 mRNA and protein expression of different concentrations of STS intervention group〔0.588±0.077,(61.922±5.821)ng/ml〕、〔0.530±0.070,(57.976±8.009)ng/ml〕、〔0.222±0.029,(43.291±1.878)ng/ml〕were decreased significantly ,but still higher than control group(P<0.01, respectively). Compared with nicotine group, the t-PA mRNA expression of 20μmol/L STS group(0.184±0.023)were increased not significantly(P>0.05);The t-PA mRNA expression of 50、100μmol/L STS group(0.293±0.037)、(0.412±0.053)were increased significantly ,but still lower than control group(P<0.01, respectively).The expression of t-PA protein was no significantly different (P>0.05).
3.Compared with nicotine group , the PAI-1 mRNA and protein expression of different concentrations of simvastatin intervention group〔0.581±0.095,(60.788±2.500)ng/ml〕、〔0.511±0.084,(54.955±3.971)ng/ml〕、〔1.074±0.099,(16.190±2.149)ng/ml〕were decreased significantly ,and the most notably effect of simvastatin was at the concentrations of 100μmol/L ,almost the same as that in the control group(P>0.05). Compared with nicotine group, the t-PA mRNA expression of 1μmol/L simvastatin group(0.187±0.025)were increased not significant(P>0.05);The t-PA mRNA expression of 10、100μmol/L simvastatin group(0.278±0.038)、(0.332±0.046)were increased significantly ,but still lower than control group(P<0.01, respectively). The expression of t-PA protein was no significantly different (P>0.05).
Conclusions
1.Nicotine through increased the expression of PAI-1 protein and mRNA in HUVECs,inhibits the fibrinolytic function of endothelial cells.
2.Activation the PKC can partly disorder the fibrinolytic function of endothelial cells that induced by nicotine.
3.Simvastatin can improves the fibrinolytic function of endothelial cells that induced by nicotine
引文
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[11]Catley MC, Cambridge LM, Nasuhara Y, et al. Inhibitors of protein kinase C (PKC) prevent activated transcription.J Biol Chem,2004,279:18457-18466.
[12]Housey GM, Johnson MD, Wendy HW, et al. Overproduction of protein kinase C causes disorderd growth control in rat fibroblasts.Cell,1988,52(3):343-348.
[13]Takahashi I, Kobayashi E, Nakano H, et al. Potent selective inhibition of 7-0-methyl UCN-01 against protein kinase C.J Pharmacol Exp Ther,1990,255(3):1218-1221.
[14]Banfi C, Mussoni L, Rise P. Very low density lipoprotein-mediated signal transduction and plasminogen activator inhibitor type 1 in culture Hep G2 cells. Circ Res,1999,85(20:208-217.
[15]Lee S D, Lee D S, Chun Y G, et al. Cigarette smoke extract induces endothelin-1 via protein kinase C in pulmonary artery endothelial cells.Am J Physiol Lung Cell MolPhysiol,2001,281:L403-L411.
[16]Conklin BS et al.Am J Pathol,2002,160:413
[17]杨霄鹏,李秋芳,王金兰,等.急性脑梗死患者血浆t-PA和PAI的变化及临床意义.中国实用神经疾病杂志,2006,9(6):33～34.
[18]扬中苏,杨新春,那开宪,等.他汀类药物的临床应用进展.世界急危重病医学杂志,2006,3(5):1496-1502.
[19]Sadeghi MM, Collinge M, Pardi R, et al. Simvastatin modulates cytokine-mediated endothelial cell adhesion molecule induction:involvement of an inhibitory G-protein.J Immunology,2000,165(5):2712-2718.
[20]Ye P , Hu X , Zhao Y . The increase in plasminogen activator inhibitor type-1 expression by stimulation of activators for peroxisome proliferator-activated receptors in human endothelial cells.Chin Med Sci.2002 Jun;17(2):112-116.
[21]杜艳,胡晓芸,杜永成,等.尼古丁对血管内皮细胞释放t-PA及PAI-1的影响.中国病理生理杂志,2010,26(11):2253-2255.
[22]Wiesbauer F , Kaun C , Zorn G ,et al. HMG CoA reductase inhibitors affect the fibrenolytic system of human vascular cells in vitro:a comparative study using different statins.Br J Pharamcol.2002 Jan;135(1):284-292.
[23]Veillard NR , Braunersreuther V , Arnaud C , et al. Simvastatin modulates chemokine and chemokine receptor expression by geranylgeranyl isoprenoid pathway in human endothelial cells and macrophages. Atherosclerosis.2006 Sep;188(1):51-58.
[24]Kaftan AH , Kaftan O.Coronary artery disease and infection with chlamydia pneumonia.Jpn Heart J.2000 Mar;41(2):165-172.
[25]Grulich-Henn J , Muller-Berghaus G .Regulation of endothelial tissue plasminogen activator and plasminogen activator inhibitor type 1 syntheses by diacylglycerol,phorbol ester and thrombin.Blut,1990 Jul;61(1):38-44.
[26]刘永平,郑兴,秦永文.辛伐他汀对冠心病患者血浆纤溶活性的改善作用.实用新医药杂志,2006,23(5):539-541.
[27]FUL , Jin H , Song K ,et al. Relationship between gene polymorphism of the PAI-1 promoter and myocardial infarction.Chin Med J,2001 Mar;114(3):266-269.
[28]Tsantes AE , Nikolopoulos GK , Bagos PG , et al. Association between the plasminogenactivator inhibitor-1 4G/5G polymorphism and venous thrombosis. A meta-analysis.Thromb Haemost.2007 Jun; 97 (6):907-913.
[1]Azzi A, Boscoboinik D, Hensey C. The protein kinase C family. Eur J Biochem, 1992, 208 (3):547-557.
[2]House C, Kemp BE. Protein kinase C contains a pseudosubstrate peototope in its regulatory domain .Science,1987,238(4834):1726-1728.
[3]Ward JP,Knock GA, Snetkov VA, Aaronson P I Protein kinases in vascular smooth muscle tone-role in the pulmonary vasculature and hypoxic pulmonary vasoconstriction. Pharmacology Therapeutics,2004,104(3):207-31.
[4]杨滔,黄巧冰.蛋白激酶C在调节血管内皮通透性中的研究.中国病理生理杂志,2002,18 (6):722-4.
[5]Nishizuka Y. Studies and prospectives of the protein kinase C family for cellular regulation.Cancer,1989,63(10):1892-1903.
[6]Richard M.膜的生物物理性质在调节蛋白激酶活性中的作业.国外医学:医学分册, 1991,18(3):169-171.
[7]Loscalzo J, Welch G Nitric oxide and its role in the cardiovascular system. Prog Cardiovasc D is,1995,38(2):87.
[8]Antoniades C, Tousoulis D, Vasiliadou C, et al. Genetic polymorphism on endothelial nitric oxide synthase affects endothelial activation and inflammatory response during the acute phase of myocardial infarction J Am Coll Cardiol,2005,46(6):1101.
[9]Wedgwood S, Bekker JM, Black SM.Shear stress regulation of endothelial NOS in fetal pulmonary arterial endothlial cells involves PKC.Am J Physiol Lung Cell Mol Physiol,2001,281(2):L490-498.
[10]Feletou M, Vanhoutte PM. Endothelial dysfunction: a multifaceted disorder.Am J Physiol Heart Circ Physiol,2006,291:H985-1002.
[11]Markovk HM. Molecular mechanisms of dysfunction of vascular endothelium. Kardiologiia ,2005,45:62-72.
[12]Yakubu MA, Leffler CW. Regulation of ET-1 biodynthesis in cerebral microvascular endothelial cells by vasoactive agents and PKC.Am J Physiol Cell Physiol,1999,276(2 Pt 1):C300-C305.
[13]Sarker R, Meinborg EG, Standery JC, et al.Nitric oxide reversibly inhibits the migration of cultured vascular smooth muscle cells.Circ Res,1996,78(3):225.
[14]Graham MA, Rawe I, Dartt DA, et al. Protein kinase C regulation of corneal endothelial cell proliferation and cell cycle.Invest Ophthalmol Vis Sci, 2000, 41 (13):4124-4132.
[15]Wang JH, Redmond HP, Watson RW, et al. Involvement of tyrosine protein kinase in IFN-gamma-induced human endothelial cell apoptosis. Shockm 1999, 11 (5): 311 - 318.
[16]Schwartz GK, Haimovitz-Friedman A. Dhupar SK,et al.Potentiation of apotosis by treatment with protein kinase C special inhibitor safingol in Mitomyein C treated gastric eancer cells.J Nati Cancer Inst.1995.87:1394-1399.
[17]Dumont JA, Bitonti AJ. Modulation of human melanoma cell metastasis and adhesion may involve integrin phosphorylation mediated through protein kinase C.Biochem biophys Res Commun,1994,204(1):264.
[18]Haier J, Nasralla MY, Nicolson GL. Betal-integrin-mediated dynamic adhesion of colon carcinoma cells to extra-cellular matrix under laminar flow.Clin Exp Metastasis , 1999,17(5):377.
[19]Chmura SJ, Mauceri HJ, Advani S, et al. Decreasing the apoptotic threshold of tumor cells through protein kinase C inhibitor and sphingomyelinase activation increases tumor killing by ionizing radiation.Cancer Res,1997,57:4540-4544.
[20]Smith CW. Possible steps involved in the transition to stationary adhesion of rolling nertrophils:a brief review,Microcirculation.2000;7:385-394.
[21]Springer TA. Traffic signals for lymphocyte recirculation and leukocyte emigration:the multistep paradigm.Cell,1994;76:301-314.
[22]Serfilippi G, Ferro TJ, Johnson A. Activation of protein kianse C mediates altered pulmonary vasoreactivity induced by tumor necrosis factor-alpha.Am J Physiol,1994,267:282-290.
[23]Zachary L, Mathur A, Yla-Herttuala S, et al. Vascular protection:A novel,non-angiogenic cardiovascular role for vascular endothelial growth factor.Arterioscler Thromb Vasc Biol,2000,20:1512-1520.
[24]Lucerna M, Zernecke A, de-Nooijer R, et al. Vascular endothelial growth factor-A induces plaque expansion in Apo E knock-out mice by promoting novo leukocyte recruitment .Blood,2007,109:122-129.
[25]Deem TL, Abdala-Valencia H, Cook-Mills JM.VCAM-1 activation of endothelial cell protein tyrosine phosphatase IB.J Immunol,2007,178:3865-3873.
[26]李鹏程,徐茜,胡艳华.蛋白激酶C对SDF-1诱导巨噬细胞趋化及血管内皮生长因子转录的调节作用.华中科技大学学报(医学版).2008,37(1):93-96.
[27]黄巧冰,杨滔,宋力,等.蛋白激酶C在调节血管内皮细胞通透性中的作用.中国病理生理杂志,2001,17(11):1138-1139.
[28]郝一文,丁伟,刘显智.蛋白激酶C对血管内皮功能的影响.医学临床研究,2006,23(6):828-830.
[29]Lum H, Malik AB. Regulation of vascular endothelial barrier function.Am J Physiol, 1994,267:2232241.
[30]Banfi C, Mussoni L, Rise P. Very low density lipoprotein-mediated signal transduction and plasminogen activator inhibitor type 1 in culture HepG2 cells.Circ Res,1999,85(2):208-217.
[31]Guey-Yueh S, Shwu-jyh W, Chi-Fong Tasi. Regulation of plasminogen activator inhibitor activity by plasmin in endothelial cells. Thromb Res,1996,81(1):75-84.
[32]Grulich-Henn J, Muller-Berghaus G. Regulation of endothelial tissue plasminogen activator and plasminogen activator inhibitor type 1 syntheses by diacylglycerol,phorbol ester and thrombin. Blut, 1990,61(1):38-44.
[33]Niedbala MJ, Stein-Picarella M. Role of protein kinase C in tumor factor induction of endothelial cell urokinase plasminogen activator.Blood,1993,81(10):2608-2617.
[34]Zidovetzki R , Wang JL ,Kim JA ,et al.Endothelin-1 enhances plasminogen activator inhibitor-1 production by human brain endothelial cells via protein kinase C-dependent pathway . Arterioscler Thromb Vasc Biol,1999,19(7):1768-1775.
[2]Schondorf T, Hoopmann M, Breidenbach M, et al. Dysregulation of protein kinase C activity in chemoresistant metastatic breast cancer cells. Anticancer Drugs,2004, 15(3):265-268.
[3]Yanyan Wang,Hongfa Yang,Hongbin Liu,et al. Effect of staurosporine on the mobility and invasiveness of lung adenocarcinoma A549 cells: an in vitro study. BMC Cancer, 2009, 9:174.
[4]Smith EB. Haemostatic factor and atherogenesis.Atherosclerosis,1996,124(2):137-143.
[5]Victor WM, Kooistra T. Tumor necrosis factor increases the production of plasminogen activator inhibitor in human endothelial cells in vitro and rats in vivo. Blood, 1998, 72(5): 1467-1473.
[6]David J, Schneider. Changes in arterial expression of fibrinolytic system protein in atherogenesis.Arterios Thromb Vasc Biol,1997,17(11):3294-3301.
[7]Guey-Yueh S, Shwu-jyh W, Chi-Fong Tasi. Regulation of plasminogen activator inhibitor activity by plasmin in endothelial cells.Thromb Res,1996,81(1):75-84.
[8]Grulich-Henn J, Muller-Berghaus G. Regulation of endothelial tissue plasminogen activator and plasminogen activator inhibitor type 1 syntheses by diacylglycerol, phorbol ester and thrombin.Blut,1990,61(1):38-44.
[9]Niedbala MJ, Stein-Picarella M. Role of protein kianse C in tumor factor induction of endothelial cell urokinase plasminogen activator.Blood,1993,81(10):2608-2617.
[10]Michic AM, Nakagawa R. The link between PKC-αregulation and cellular transformation.Immunol Letters,2005,96:155-162.
[11]Catley MC, Cambridge LM, Nasuhara Y, et al. Inhibitors of protein kinase C (PKC) prevent activated transcription.J Biol Chem,2004,279:18457-18466.
[12]Housey GM, Johnson MD, Wendy HW, et al. Overproduction of protein kinase C causes disorderd growth control in rat fibroblasts.Cell,1988,52(3):343-348.
[13]Takahashi I, Kobayashi E, Nakano H, et al. Potent selective inhibition of 7-0-methyl UCN-01 against protein kinase C.J Pharmacol Exp Ther,1990,255(3):1218-1221.
[14]Banfi C, Mussoni L, Rise P. Very low density lipoprotein-mediated signal transduction and plasminogen activator inhibitor type 1 in culture Hep G2 cells. Circ Res,1999,85(20:208-217.
[15]Lee S D, Lee D S, Chun Y G, et al. Cigarette smoke extract induces endothelin-1 via protein kinase C in pulmonary artery endothelial cells.Am J Physiol Lung Cell MolPhysiol,2001,281:L403-L411.
[16]Conklin BS et al.Am J Pathol,2002,160:413
[17]杨霄鹏,李秋芳,王金兰,等.急性脑梗死患者血浆t-PA和PAI的变化及临床意义.中国实用神经疾病杂志,2006,9(6):33～34.
[18]扬中苏,杨新春,那开宪,等.他汀类药物的临床应用进展.世界急危重病医学杂志,2006,3(5):1496-1502.
[19]Sadeghi MM, Collinge M, Pardi R, et al. Simvastatin modulates cytokine-mediated endothelial cell adhesion molecule induction:involvement of an inhibitory G-protein.J Immunology,2000,165(5):2712-2718.
[20]Ye P , Hu X , Zhao Y . The increase in plasminogen activator inhibitor type-1 expression by stimulation of activators for peroxisome proliferator-activated receptors in human endothelial cells.Chin Med Sci.2002 Jun;17(2):112-116.
[21]杜艳,胡晓芸,杜永成,等.尼古丁对血管内皮细胞释放t-PA及PAI-1的影响.中国病理生理杂志,2010,26(11):2253-2255.
[22]Wiesbauer F , Kaun C , Zorn G ,et al. HMG CoA reductase inhibitors affect the fibrenolytic system of human vascular cells in vitro:a comparative study using different statins.Br J Pharamcol.2002 Jan;135(1):284-292.
[23]Veillard NR , Braunersreuther V , Arnaud C , et al. Simvastatin modulates chemokine and chemokine receptor expression by geranylgeranyl isoprenoid pathway in human endothelial cells and macrophages. Atherosclerosis.2006 Sep;188(1):51-58.
[24]Kaftan AH , Kaftan O.Coronary artery disease and infection with chlamydia pneumonia.Jpn Heart J.2000 Mar;41(2):165-172.
[25]Grulich-Henn J , Muller-Berghaus G .Regulation of endothelial tissue plasminogen activator and plasminogen activator inhibitor type 1 syntheses by diacylglycerol,phorbol ester and thrombin.Blut,1990 Jul;61(1):38-44.
[26]刘永平,郑兴,秦永文.辛伐他汀对冠心病患者血浆纤溶活性的改善作用.实用新医药杂志,2006,23(5):539-541.
[27]FUL , Jin H , Song K ,et al. Relationship between gene polymorphism of the PAI-1 promoter and myocardial infarction.Chin Med J,2001 Mar;114(3):266-269.
[28]Tsantes AE , Nikolopoulos GK , Bagos PG , et al. Association between the plasminogenactivator inhibitor-1 4G/5G polymorphism and venous thrombosis. A meta-analysis.Thromb Haemost.2007 Jun; 97 (6):907-913.
[1]Azzi A, Boscoboinik D, Hensey C. The protein kinase C family. Eur J Biochem, 1992, 208 (3):547-557.
[2]House C, Kemp BE. Protein kinase C contains a pseudosubstrate peototope in its regulatory domain .Science,1987,238(4834):1726-1728.
[3]Ward JP,Knock GA, Snetkov VA, Aaronson P I Protein kinases in vascular smooth muscle tone-role in the pulmonary vasculature and hypoxic pulmonary vasoconstriction. Pharmacology Therapeutics,2004,104(3):207-31.
[4]杨滔,黄巧冰.蛋白激酶C在调节血管内皮通透性中的研究.中国病理生理杂志,2002,18 (6):722-4.
[5]Nishizuka Y. Studies and prospectives of the protein kinase C family for cellular regulation.Cancer,1989,63(10):1892-1903.
[6]Richard M.膜的生物物理性质在调节蛋白激酶活性中的作业.国外医学:医学分册, 1991,18(3):169-171.
[7]Loscalzo J, Welch G Nitric oxide and its role in the cardiovascular system. Prog Cardiovasc D is,1995,38(2):87.
[8]Antoniades C, Tousoulis D, Vasiliadou C, et al. Genetic polymorphism on endothelial nitric oxide synthase affects endothelial activation and inflammatory response during the acute phase of myocardial infarction J Am Coll Cardiol,2005,46(6):1101.
[9]Wedgwood S, Bekker JM, Black SM.Shear stress regulation of endothelial NOS in fetal pulmonary arterial endothlial cells involves PKC.Am J Physiol Lung Cell Mol Physiol,2001,281(2):L490-498.
[10]Feletou M, Vanhoutte PM. Endothelial dysfunction: a multifaceted disorder.Am J Physiol Heart Circ Physiol,2006,291:H985-1002.
[11]Markovk HM. Molecular mechanisms of dysfunction of vascular endothelium. Kardiologiia ,2005,45:62-72.
[12]Yakubu MA, Leffler CW. Regulation of ET-1 biodynthesis in cerebral microvascular endothelial cells by vasoactive agents and PKC.Am J Physiol Cell Physiol,1999,276(2 Pt 1):C300-C305.
[13]Sarker R, Meinborg EG, Standery JC, et al.Nitric oxide reversibly inhibits the migration of cultured vascular smooth muscle cells.Circ Res,1996,78(3):225.
[14]Graham MA, Rawe I, Dartt DA, et al. Protein kinase C regulation of corneal endothelial cell proliferation and cell cycle.Invest Ophthalmol Vis Sci, 2000, 41 (13):4124-4132.
[15]Wang JH, Redmond HP, Watson RW, et al. Involvement of tyrosine protein kinase in IFN-gamma-induced human endothelial cell apoptosis. Shockm 1999, 11 (5): 311 - 318.
[16]Schwartz GK, Haimovitz-Friedman A. Dhupar SK,et al.Potentiation of apotosis by treatment with protein kinase C special inhibitor safingol in Mitomyein C treated gastric eancer cells.J Nati Cancer Inst.1995.87:1394-1399.
[17]Dumont JA, Bitonti AJ. Modulation of human melanoma cell metastasis and adhesion may involve integrin phosphorylation mediated through protein kinase C.Biochem biophys Res Commun,1994,204(1):264.
[18]Haier J, Nasralla MY, Nicolson GL. Betal-integrin-mediated dynamic adhesion of colon carcinoma cells to extra-cellular matrix under laminar flow.Clin Exp Metastasis , 1999,17(5):377.
[19]Chmura SJ, Mauceri HJ, Advani S, et al. Decreasing the apoptotic threshold of tumor cells through protein kinase C inhibitor and sphingomyelinase activation increases tumor killing by ionizing radiation.Cancer Res,1997,57:4540-4544.
[20]Smith CW. Possible steps involved in the transition to stationary adhesion of rolling nertrophils:a brief review,Microcirculation.2000;7:385-394.
[21]Springer TA. Traffic signals for lymphocyte recirculation and leukocyte emigration:the multistep paradigm.Cell,1994;76:301-314.
[22]Serfilippi G, Ferro TJ, Johnson A. Activation of protein kianse C mediates altered pulmonary vasoreactivity induced by tumor necrosis factor-alpha.Am J Physiol,1994,267:282-290.
[23]Zachary L, Mathur A, Yla-Herttuala S, et al. Vascular protection:A novel,non-angiogenic cardiovascular role for vascular endothelial growth factor.Arterioscler Thromb Vasc Biol,2000,20:1512-1520.
[24]Lucerna M, Zernecke A, de-Nooijer R, et al. Vascular endothelial growth factor-A induces plaque expansion in Apo E knock-out mice by promoting novo leukocyte recruitment .Blood,2007,109:122-129.
[25]Deem TL, Abdala-Valencia H, Cook-Mills JM.VCAM-1 activation of endothelial cell protein tyrosine phosphatase IB.J Immunol,2007,178:3865-3873.
[26]李鹏程,徐茜,胡艳华.蛋白激酶C对SDF-1诱导巨噬细胞趋化及血管内皮生长因子转录的调节作用.华中科技大学学报(医学版).2008,37(1):93-96.
[27]黄巧冰,杨滔,宋力,等.蛋白激酶C在调节血管内皮细胞通透性中的作用.中国病理生理杂志,2001,17(11):1138-1139.
[28]郝一文,丁伟,刘显智.蛋白激酶C对血管内皮功能的影响.医学临床研究,2006,23(6):828-830.
[29]Lum H, Malik AB. Regulation of vascular endothelial barrier function.Am J Physiol, 1994,267:2232241.
[30]Banfi C, Mussoni L, Rise P. Very low density lipoprotein-mediated signal transduction and plasminogen activator inhibitor type 1 in culture HepG2 cells.Circ Res,1999,85(2):208-217.
[31]Guey-Yueh S, Shwu-jyh W, Chi-Fong Tasi. Regulation of plasminogen activator inhibitor activity by plasmin in endothelial cells. Thromb Res,1996,81(1):75-84.
[32]Grulich-Henn J, Muller-Berghaus G. Regulation of endothelial tissue plasminogen activator and plasminogen activator inhibitor type 1 syntheses by diacylglycerol,phorbol ester and thrombin. Blut, 1990,61(1):38-44.
[33]Niedbala MJ, Stein-Picarella M. Role of protein kinase C in tumor factor induction of endothelial cell urokinase plasminogen activator.Blood,1993,81(10):2608-2617.
[34]Zidovetzki R , Wang JL ,Kim JA ,et al.Endothelin-1 enhances plasminogen activator inhibitor-1 production by human brain endothelial cells via protein kinase C-dependent pathway . Arterioscler Thromb Vasc Biol,1999,19(7):1768-1775.