地塞米松对大鼠胸膜间皮细胞水通道蛋白1表达的调节  

Effect of dexamethasone on the expression of aquaporin-1 in cultured rat pleural mesothelial cells

作　　者：张伟[1] 谢灿茂[1] 何桥[2] 邓晓华[2] 柯彩霞[1] 

机构地区：[1]中山大学附属第一医院呼吸内科,广东广州510080 [2]广州市胸科医院病理科,广东广州510095

出　　处：《中国病理生理杂志》2007年第1期22-26,共5页Chinese Journal of Pathophysiology

基　　金：教育部博士点基金资助项目(No.20050558060);中国博士后科学基金资助项目(No.2005038182)

摘　　要：目的:观察胸膜间皮细胞水通道蛋白1(AQP-1)的表达及地塞米松(Dex)对其表达的影响,为进一步研究胸水治疗的机制提供实验依据。方法:体外培养大鼠胸膜间皮细胞,细胞鉴定后,采用免疫细胞化学、RT-PCR方法检测AQP-1表达;应用Westernblotting检测以不同浓度Dex(10-8、10-7、10-6、10-5、10-4mmol/L)处理细胞24h及以10-4mmol/LDex处理细胞6h、12h、24h、36h、48h、72h后AQP-1表达情况。结果:发现胸膜间皮细胞存在AQP-1表达,10-8、10-7、10-6、10-5、10-4mmol/LDex干预胸膜间皮细胞24h后,AQP-1蛋白表达量(积分吸光度)分别为755.04±19.81、843.72±19.41、862.96±26.53、694.80±32.00、938.08±13.32,分别高于正常对照组(372.90±16.46)2.02、2.26、2.31、1.86、2.52倍(P<0.01),AQP-1表达升高与地塞米松浓度无关;以10-4mmol/LDex干预细胞6h、12h、24h、36h、48h、72h后,AQP-1蛋白表达量分别为:554.14±23.57、917.78±38.62、1587.20±61.22、1322.09±28.65、918.40±26.62、1117.60±51.32,分别高于正常对照组(495.91±23.12)1.12、1.85、3.20、2.67、1.85、2.25倍(P<0.01),AQP-1表达与地塞米松作用时间有关。结论:胸膜间皮细胞存在AQP-1表达,地塞米松对胸膜间皮细胞AQP-1表达有明显的增强性调节作用,具有时间依赖性。AIM： To study the effect of dexamethasone （Dex） on the expression of aquaporin - 1 （ AQP - 1 ） in cultured rat pleural mesothelial cells and to offer experimental support for the investigation of the mechanisms of pleural fluid treatment. METHODS： Rat pleural mesothefial cells were cultured in vitro. The expression of AQP- 1 was detected by immunocytochemistry and reverse transcription - PCR （ RT - PCR） after cells were identified, then the cens were treated with Dex for24 hours at concentrations of 10^-8、10^-7、10^-5 and 10^-4mmol/L, and for6 h, 12 h, 24 h, 36 h, 48 h, 72 h at concentration of 10^-4 mmoL/L. Western blotting was used to analyze the expression of AQP - 1 in cultured control and Dex--treated rat pleural mesothelial cells, and image analysis system to analyze the expression of AQP - 1. RESULTS： Aquaporin- 1 was expressed in cultured rat pleural mesothelial cells. The protein of.AQP- 1 expressed in rat pleurai mesothelial cells was 755.04 ± 19. 81, 843.72 ± 19.41, 862.96 ± 26. 53, 694. 80 ± 32.00, 938.08 ± 13.32 in those treated with Dex at concentrations of 10-s, 10-7, 10＂±, 10-5 and 10-4 mmol/L, respectively, the levels was 2. 02, 2. 26, 2. 31, 1.86, 2.52 fold higher than that in control group （372. 90 ± 16. 46, P 〈0. 01 ）. The protein of AQP - 1 ex- pressed was 5.54. 14±23.57, 917.78 ±38.62, 1 587.20 ±61.22, 1 322.09 ±28.65, 918.40 ±26.62, 1 117.60 ± 51.32 in those treated with 10-4 mmol/L Dex after 6 h, 12 h, 24 h, 36 h, 48 h, 72 h, the levels was 1.12, 1.85, 3.20, 2. 67, 1.85 and 2. 2.5 fold higher than that in control group （495.91 ±23. 12, P〈0. 01）. CONCLUSION： The expression of AQP - 1 in cultured rat pleural mesothefial cells is confirmed at the protein and mRNA level. Dex increases the expression of AQP - 1 in a time - dependent manner.

关 键 词：地塞米松 胸膜间皮细胞 水孔蛋白类 

分 类 号：R363[医药卫生—病理学]