国际呼吸杂志
國際呼吸雜誌
국제호흡잡지
INTERNATIONAL JOURNAL OF RESPIRATION
2012年
24期
1860-1863,封3
,共5页
李爱民%扈丽娟%张医婵%刘志宏%胡晓芸
李愛民%扈麗娟%張醫嬋%劉誌宏%鬍曉蕓
리애민%호려연%장의선%류지굉%호효예
Staurosporine%尼古丁%人脐静脉内皮细胞%组织型纤溶酶原激活物%1型纤溶酶原激活物抑制剂
Staurosporine%尼古丁%人臍靜脈內皮細胞%組織型纖溶酶原激活物%1型纖溶酶原激活物抑製劑
Staurosporine%니고정%인제정맥내피세포%조직형섬용매원격활물%1형섬용매원격활물억제제
Staurosporine%Nicotine%Human umbilical vein endothelial cells%Tissue type plasminogen activator%Plasminogen activator inhibitor-1
目的 通过观察不同浓度Staurosporine (STS)对尼古丁诱导人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)表达组织型纤溶酶原激活物(tissue type plasminogen activator,t-PA)与1型纤溶酶原激活物抑制剂(plasminogen activator inhibitor-1,PAI-1)mRNA及蛋白的影响,探讨尼古丁所致内皮细胞纤溶紊乱的机制.方法 将体外培养的3～6代HUVECs随机分为对照组、尼古丁组及不同浓度STS组,STS组分别以20、50、100μmol/L STS预处理细胞30 min,再与100 μmol/L尼古丁孵育24 h.酶联免疫吸附双抗体夹心法检测细胞上清液t-PA与PAI-1蛋白含量,RT-PCR检测PAI-1 mRNA表达.结果 尼古丁组PAI-1 mRNA与蛋白表达较对照组升高(P值均＜0.05)；不同浓度STS组PAI-1 mRNA与蛋白表达较尼古丁组降低(P值均＜0.05),且呈浓度依赖性,以100 μmol/L STS组作用为著,但PAI-1 mRNA与蛋白表达仍高于对照组(P值均＜0.05).各组间t-PA蛋白差异无统计学意义(P值均＞0.05).结论 STS通过阻断蛋白激酶C通路的信号传导可部分减弱尼古丁诱导的血管内皮细胞纤溶功能紊乱.
目的 通過觀察不同濃度Staurosporine (STS)對尼古丁誘導人臍靜脈內皮細胞(human umbilical vein endothelial cells,HUVECs)錶達組織型纖溶酶原激活物(tissue type plasminogen activator,t-PA)與1型纖溶酶原激活物抑製劑(plasminogen activator inhibitor-1,PAI-1)mRNA及蛋白的影響,探討尼古丁所緻內皮細胞纖溶紊亂的機製.方法 將體外培養的3～6代HUVECs隨機分為對照組、尼古丁組及不同濃度STS組,STS組分彆以20、50、100μmol/L STS預處理細胞30 min,再與100 μmol/L尼古丁孵育24 h.酶聯免疫吸附雙抗體夾心法檢測細胞上清液t-PA與PAI-1蛋白含量,RT-PCR檢測PAI-1 mRNA錶達.結果 尼古丁組PAI-1 mRNA與蛋白錶達較對照組升高(P值均＜0.05)；不同濃度STS組PAI-1 mRNA與蛋白錶達較尼古丁組降低(P值均＜0.05),且呈濃度依賴性,以100 μmol/L STS組作用為著,但PAI-1 mRNA與蛋白錶達仍高于對照組(P值均＜0.05).各組間t-PA蛋白差異無統計學意義(P值均＞0.05).結論 STS通過阻斷蛋白激酶C通路的信號傳導可部分減弱尼古丁誘導的血管內皮細胞纖溶功能紊亂.
목적 통과관찰불동농도Staurosporine (STS)대니고정유도인제정맥내피세포(human umbilical vein endothelial cells,HUVECs)표체조직형섬용매원격활물(tissue type plasminogen activator,t-PA)여1형섬용매원격활물억제제(plasminogen activator inhibitor-1,PAI-1)mRNA급단백적영향,탐토니고정소치내피세포섬용문란적궤제.방법 장체외배양적3～6대HUVECs수궤분위대조조、니고정조급불동농도STS조,STS조분별이20、50、100μmol/L STS예처리세포30 min,재여100 μmol/L니고정부육24 h.매련면역흡부쌍항체협심법검측세포상청액t-PA여PAI-1단백함량,RT-PCR검측PAI-1 mRNA표체.결과 니고정조PAI-1 mRNA여단백표체교대조조승고(P치균＜0.05)；불동농도STS조PAI-1 mRNA여단백표체교니고정조강저(P치균＜0.05),차정농도의뢰성,이100 μmol/L STS조작용위저,단PAI-1 mRNA여단백표체잉고우대조조(P치균＜0.05).각조간t-PA단백차이무통계학의의(P치균＞0.05).결론 STS통과조단단백격매C통로적신호전도가부분감약니고정유도적혈관내피세포섬용공능문란.
Objective To detect the mechanism of the effects of different concentrations of Staurosporine on nicotine induced expression of tissue type plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1) at mRNA and protein levels in human umbilical vein endothelial cells (HUVECs).Methods Cultured HUVECs at passage 3 to 6 were divided into the control group,nicotine group,various-dose Staurosporine groups.After intervented by different concentrations (20,50,100 μ mol/L) of Staurosporine for 30 minutes,the various dose Staurosporine groups were stimulated by nicotine (100 μmol/L) for 24 hours.The protein levels of t-PA and PAI-1 in the cultured medium were measured by enzyme-linked immunosorbent assay.PAI-1 mRNA level was assayed by reverse transcript-polymerase chain reaction.Results Compared to control group,the expressions of PAI-1 mRNA and protein increased in nicotine group (all P ＜0.05).Compared to nicotine group,the expressions of PAI-1 mRNA and protein in various-dose Staurosporine groups decreased by means of dose-dependent,the effect of 100 μmol/L Staurosporine was notable,but the expressions of PAI-1 mRNA and protein were higher than those in control group (all P ＜0.05).However,the expression of t-PA antigens had no significant change among all groups (all P ＞0.05).Conclusions Through inhibiting the activation of PKC signaling pathway, Staurosporine partly improves the fibrinolytic function of HUVECs induced by nicotine in vitro.
