[蛋白激酶C-细胞外信号调节激酶1/2信号通路在尼古丁诱导人脐静脉内皮细胞纤溶酶原激活物抑制剂-1表达过程中的相互作用]

[Interaction of protein kinase C-extracellular signal-regulated kinase 1/2 signal pathway in the process of plasminogen activator inhibitor-1 expression induced by nicotine in human umbilical vein endothelial cells].

作者信息

DU Yan, Hu Xiao-yun, Han Bao-fen, DU Yong-cheng

机构信息

Department of Respiratory Medicine, First Hospital, Shanxi Medical University, Taiyuan 030001, China.

出版信息

Zhonghua Jie He He Hu Xi Za Zhi. 2011 Jun;34(6):442-6.

PMID:21781517

Abstract

OBJECTIVE

To explore the role of protein kinase C (PKC)- extracellular signal-regulated kinase (ERK)1/2 signal pathway in the process of plasminogen activator inhibitor-1 (PAI-1) protein and mRNA expression in cultured human umbilical vein endothelial cells (HUVECs) induced by nicotine.

METHODS

HUVECs were cultured to examine the effect of nicotine on the expression of secreting PAI-1 in HUVECs on different experimental conditions. The expression of PAI-1 protein was measured by ELISA. PKC inhibitor staurosporine (STS) and ERK1/2 inhibitor PD98059 were used to detect PKC or ERK1/2 function on the expression of PAI-1 in HUVECs induced by nicotine. The PAI-1 mRNA expression was determined by RT-PCR.

RESULTS

The expression level of PAI-1 protein in 100 µmol/L nicotine treated group [(22.6 ± 1.1) µg/L] increased significantly compared to the control group [(14.2 ± 2.8) µg/L; q = 5.64, P < 0.05]. After stimulation with 100 µmol/L nicotine for 0, 4, 6, 8, 12 and 24 h, the levels of PAI-1 protein increased over time and reached the peak at 12 h (F = 32.063, P < 0.05). The PAI-1 mRNA and protein expression in nicotine treated group [(1.32 ± 0.20), (21.08 ± 0.83) µg/L] increased significantly compared to the control group [(0.73 ± 0.10), (13.39 ± 0.93) µg/L; q = 8.43, 11.97, all P < 0.05].Compared with nicotine treated group, the PAI-1 mRNA and protein expression in nicotine and STS treated group [(1.07 ± 0.10), (16.19 ± 2.15) µg/L] decreased significantly(q = 5.61, 7.61, all P < 0.05), but still higher than the control group (q = 7.84, 4.36, all P < 0.05). In nicotine and PD98059 treated group, the PAI-1 mRNA and protein expression [(1.12 ± 0.11), (17.52 ± 1.72) µg/L] decreased significantly compared to the nicotine treated group(q = 4.68, 5.54, all P < 0.05), still higher than the control group (q = 8.77, 6.43, all P < 0.05).

CONCLUSION

PKC-ERK1/2 signal pathway may play a partial role in the up-regulation of PAI-1 induced by nicotine in HUVECs.

摘要

目的

探讨蛋白激酶C（PKC）-细胞外信号调节激酶（ERK）1/2信号通路在尼古丁诱导的人脐静脉内皮细胞（HUVECs）纤溶酶原激活物抑制剂-1（PAI-1）蛋白及mRNA表达过程中的作用。

方法

培养HUVECs，检测尼古丁在不同实验条件下对HUVECs分泌PAI-1表达的影响。采用酶联免疫吸附测定（ELISA）法检测PAI-1蛋白的表达。使用PKC抑制剂星形孢菌素（STS）和ERK1/2抑制剂PD98059检测PKC或ERK1/2功能对尼古丁诱导的HUVECs中PAI-1表达的影响。采用逆转录聚合酶链反应（RT-PCR）法检测PAI-1 mRNA的表达。

结果

100μmol/L尼古丁处理组PAI-1蛋白表达水平[（22.6±1.1）μg/L]较对照组[（14.2±2.8）μg/L]显著升高（q = 5.64，P < 0.05）。用100μmol/L尼古丁刺激0、4、6、8、12和24小时后，PAI-1蛋白水平随时间升高，并在12小时达到峰值（F = 32.063，P < 0.05）。尼古丁处理组PAI-1 mRNA和蛋白表达[（1.32±0.20），（21.08±0.83）μg/L]较对照组[（0.73±0.10），（13.39±0.93）μg/L]显著升高（q = 8.43，11.97，均P < 0.05）。与尼古丁处理组相比，尼古丁与STS处理组PAI-1 mRNA和蛋白表达[（1.07±0.10），（16.19±2.15）μg/L]显著降低（q = 5.61，7.61，均P < 0.05），但仍高于对照组（q = 7.84，4.36，均P < 0.05）。在尼古丁与PD98059处理组中，PAI-1 mRNA和蛋白表达[（1.12±0.11），（17.52±1.72）μg/L]较尼古丁处理组显著降低（q = 4.68，5.54，均P < 0.05），仍高于对照组（q = 8.77，6.43，均P < 0.05）。