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组胺对肺动脉内皮细胞一氧化氮合酶基因表达的影响

陆德琴, 李会革, 叶红, 叶仕桥, 金肆, 王迪浔

华中科技大学同济医学院, 卫生部呼吸系统疾病重点实验室. 湖北, 武汉430030

摘要

本实验研究了组胺对原代培养的肺动脉内皮细胞一氧化氮合酶(nitric oxide synthase,NOS)基因表达的影响及分子机制.采用RT-PCR和免疫印迹技术分别检测mRNA和蛋白质的表达水平,用荧光素酶报告基因实验检测eNOS基因转录起始点上游长1.6-kb的启动子活性,用硝酸还原酶法检测NO的产量.结果发现,组胺增强eNOS表达,呈浓度和时间依赖性,10μmol/L组胺处理肺动脉内皮细胞24 h可使eNOSmRNA和蛋白质的表达达到高峰,eNOSmRNA水平为正常对照组的160.8±12.2%(P＜0.05),蛋白质水平为正常对照组的136.2±11.2%(P＜0.05).特异性CaMK Ⅱ抑制剂KN-93可抑制组胺的这一效应,表明组胺可通过激活CaMK Ⅱ增强肺动脉内皮细胞eNOS基因的表达.报告基因实验表明,10μmol/L组胺处理24 h后肺动脉内皮细胞eNOS基因启动子的活性增强,为正常对照组的148.2±33.7%(P＜0.05).组胺可使肺动脉内皮细胞产生NO增加.这些结果表明组胺在转录水平增强肺动脉内皮细胞eNOS基因的表达,并使细胞产生NO增加,这可能是组胺调节肺血管张力的机制之一.CaMK Ⅱ可能是组胺增强肺动脉内皮细胞eNOS基因表达的途径之一。

关键词： 组胺; 一氧化氮合酶; 基因表达; 内皮细胞; 肺动脉

Effects of histamine on endothelial nitric oxide synthase expression inpulmonary artery endothelial cells

Lu Deqin, Li Huige, Ye Hong, ye Shiqiao, Jin Si, Wang Dixun

Department of Pathophysiology, Tongji Medical College, Huazhong University of Science and Technology, Pulmonary Disease Laboratory of Ministry of Health of China. Wuhan 430030, China

Abstract

All three nitric oxide synthase (NOS) isoforms are found in the lungs. It has been demonstrated that eNOS-derived NO plays an important role in modulating pulmonary vascular tone and inhibiting pulmonary vascular remodeling. Histamine induces pulmonary vasocon- striction by activating H_1-receptor on the smooth muscle cells and vasodilation by stimulating H_2-receptor. It remains unclear whether histamine also modulates the pulmonary vascular tone by regulating eNOS gene expression and NO production in pulmonary artery endothelial cells. Therefore, the present study was performed on cultured primary porcine pulmonary artery endothelial cells (PAECs) to investigate the effects of histamine on eNOS gene expressinn, and to explore the role of CaMK Ⅱ in eNOS gene expression. After treatment with different concentrations histamine for different times, the levels of eNOS mRNA and protein were measured by RT-PCR and Western blot, respectively.

Key words: Histamine;Nitric-oxide synthase;Gene expression;Endothelial cells;Pulmonary artery

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引用本文：

陆德琴, 李会革, 叶红, 叶仕桥, 金肆, 王迪浔. 组胺对肺动脉内皮细胞一氧化氮合酶基因表达的影响[J]. 生理学报 2004; 56 (3): .

Lu Deqin, Li Huige, Ye Hong, ye Shiqiao, Jin Si, Wang Dixun. Effects of histamine on endothelial nitric oxide synthase expression inpulmonary artery endothelial cells. Acta Physiol Sin 2004; 56 (3): (in Chinese with English abstract).