葡萄糖对大鼠胸膜间皮细胞水通道蛋白1表达的影响

Effect of Glucose on Expression of Aquaporin-1 in Pleural Mesothelial Cells in Rats

目的观察葡萄糖对胸膜间皮细胞水通道蛋白1(AQP-1)表达的影响。方法体外原代培养Wistar大鼠胸膜间皮细胞,鉴定后将细胞随机分为对照组和葡萄糖组,对照组以D-MEM/F-12培养液培养细胞,葡萄糖组以含不同浓度葡萄糖(50、100及200mmol/L)培养液培养细胞24h;100mmol/L葡萄糖组分为6、12、18及24h四个时间组,应用Western印迹、RT-PCR方法检测细胞中AQP-1表达变化。结果含50、100及200mmol/L的葡萄糖培养液作用细胞24h后,AQP-1蛋白表达量(积分吸光度)分别为54.02±4.61、127.84±9.41和231.62±22.63,分别为对照组(22.45±2.16)的2.41、5.69和10.32倍(P<0.01),AQP-1表达与葡萄糖浓度相关;以含100mmol/L葡萄糖培养液培养细胞6、12、18及24h后,AQP-1蛋白表达量分别为24.68±2.56、58.68±3.67、89.61±6.62和113.41±7.65,分别为对照组(11.81±1.45)的2.09、4.97、7.59和9.60倍,差异均有统计学意义(P<0.01),AQP-1表达与作用时间相关。AQP-1在mRNA水平表达与蛋白水平表达相一致。结论葡萄糖上调胸膜间皮细胞AQP-1的表达,具有浓度和时间相关性,AQP-1可能参与胸水的转运。

Objective To investigate the expression of aquaporin-1 （ AQP-1 ） in pleural mesothelial cells （PMCs） and the influence of glucose thereupon. Methods Rat PMCs were isolated, cultured, and divided into two groups, ie. a glucose group,cultured with glucose of different concentrations for 24 hours, and a control group,cultured in D-MEM/F-12 medium. The 100 mmol/L glucose group was administered at the time points of 6,12,18, and 24 hours respectively. RT-PCR and Western blotting were used to analyze the mRNA and protein expression of AQP-1. Results The absorbance values of AQP-1 protein expression were 54. 02 ±4. 61,127.84± 9.41, and 231.62 ± 22. 63, respectively in the PMCs treated with glucose of the concentrations of 50,100, and 200 mmol/L, all significantly higher than those in the control group （22.45± 2. 16, all P 〈 0. 01 ）. The absorbance values of AQP-1 protein expression were 24. 68 ± 2. 56,58. 68 ± 3.67, 89. 61 ±5. 62, and 113.41 ±7. 65 in the PMCs treated with glucose of the concentration of 100 mmol/L after 6,12,18 ,and 24 hours, all significantly higher than those in the control group （11.81 ± 1.45 ,P 〈 0.01 ）. Conclusions Glucose induces the expression of AQP-1 mRNA and protein. AQP-1 participates in the pleural fluid formation.