辛伐他汀对尼古丁诱导脐静脉内皮细胞分泌t-PA和PAI-1的影响

Effects of simvastatin on nicotine-induced t-PA and PAI-1 expression in human umbilical vein endothelial cells

目的:研究不同浓度辛伐他汀对尼古丁诱导人脐静脉内皮细胞(HUVECs)分泌组织纤溶酶原激活物(t-PA)和1型纤溶酶原激活物抑制剂(PAI-1)及其基因表达的影响。方法:将3-6代体外培养的HUVECs随机分为对照组、尼古丁组及不同浓度辛伐他汀组,辛伐他汀组分别以1、10、100μmol/L辛伐他汀预处理细胞2 h,再以100μmol/L尼古丁孵育24 h。酶联免疫吸附双抗体夹心法(ELISA)检测细胞上清液t-PA和PAI-1含量;逆转录聚合酶链反应(RT-PCR)检测细胞t-PA和PAI-1 mRNA的表达。结果:尼古丁组PAI-1分泌和mRNA表达较对照组显著升高(P<0.05)。不同浓度辛伐他汀组PAI-1分泌和mRNA表达均较尼古丁组显著降低,且PAI-1分泌和mRNA表达的降低呈浓度依赖性(均P<0.05),以100μmol/L辛伐他汀组最为显著。100μmol/L辛伐他汀组PAI-1分泌和mRNA表达与对照组比较,无显著差异(P>0.05)。尼古丁组t-PA mRNA表达较对照组显著降低(P<0.05)。10、100μmol/L辛伐他汀组t-PA mRNA表达较尼古丁组显著升高(P<0.05),各组间t-PA分泌无显著差异(均P>0.05)。结论:在体外,辛伐他汀可降低尼古丁所致的PAI-1分泌和mRNA的表达,并升高t-PA mRNA的表达,从而逆转尼古丁介导的HUVECs纤溶活性减低。

AIM： To investigate the effects of simvastatin on nicotine-induced expression of tissue 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 randomly divided into control group,nicotine group and simvastatin groups.The cells in simvastatin groups were treated with simvastatin at the concentrations of 1,10 and 100 μmol/L for 2 h,and then exposed to nicotine at the concentration of 100 μmol/L for 24 h.The production of t-PA and PAI-1 in the cultured medium was measured by enzyme-linked immunosorbent assay（ELISA）.The mRNA expression of t-PA and PAI-1 was determined by reverse transcript-polymerase chain reaction（RT-PCR）.RESULTS： Compared with the control cells,the secretion and mRNA expression of PAI-1 in the cells treated with nicotine were significantly increased（P〈0.05）.Simvastatin decreased the secretion and mRNA expression of PAI-1 in HUVECs stimulated by nicotine in a dose-dependent manner,and the most notable inhibitory effect was observed in 100μmol/L simvastatin group,in which the level of PAI-1 was almost the same as that in the control cells（P〈0.05）.Compared with the control cells,nicotine inhibited the mRNA expression of t-PA in HUVECs（P〈0.05）,and simvastatin at the concentrations of 10 and 100 μmol/L markedly increased the mRNA expression of t-PA in HUVECs treated with nicotine.However,the production of t-PA remained no significantly different among groups（P〉0.05）.CONCLUSION： Simvastatin obviously decreases the expression of PAI-1 at mRNA and protein levels,increases the mRNA expression of t-PA,and improves the fibrinolytic function of HUVECs exposed to nicotine in vitro.