【摘要】 为明确乙酰唑胺对水孔蛋白 1(aquaporin 1,AQP1)基因表达的抑制是否与其对细胞内pH和Ca2 + 浓度的影响有关 ,采用激光共聚焦扫描显微镜的技术观察不同培养条件下 (培养液中含或不含NaHCO3) ,10 - 5mol·L- 1 乙酰唑胺对原代培养的大鼠肾脏近曲小管上皮细胞内pH和Ca2 + 浓度的影响 ,同时用免疫荧光法观察给药 1d、3d、7d后AQP1表达的变化。结果显示在含NaHCO3组 ,乙酰唑胺处理 7d后 ,使细胞内的H+ 探针的荧光强度由 91 81± 5 4 4降至 39 5 8± 3 10 ,Ca2 + 探针荧光强度由 18 2 3± 1 0 2降至 11 5± 1 0 3,AQP1蛋白荧光强度由 4 4 4± 1 86减少到2 5 91± 1 97;在不含NaHCO3组 ,细胞内pH和Ca2 + 浓度均无明显变化 ;但AQP1蛋白的荧光强度由 4 3 15± 2 97明显减少到 2 3 85± 1 92。结果表明乙酰唑胺对细胞内pH和Ca2 + 浓度的调节依赖于细胞外液HCO3- 的存在 ,而它对AQP1表达的抑制与细胞内pH和Ca2 + 浓度的改变无关更多还原

【Abstract】 To investigate the relationship between the inhibition of aquaporin 1 (AQP1) expression by acetazolamide and the change of intracellular pH and Ca 2+ .Laser confocal scan microscopy was used to observe the effect of acetazolamide on the intracellular pH and Ca 2+ concentration in primary cultured rat renal proximal tubule epithelial cells under different culture conditions (with or without NaHCO 3 in the culture medium). The change of AQP1 expression after acetazolamide treatment was observed by immunofluorescence technology. It was shown that in NaHCO 3 group, after 7 days treatment with acetazolamide, intracellular pH signal decreased from 91.81±5.44 to 39.58±3.10, [ Ca 2+ ] i signal decreased from 18.23±1.02 to 11.5±1.03, and AQP1 protein content decreased from 44.4±1.86 to 25.91±1.97; In NaHCO 3 free group, the intracellular pH and [ Ca 2+ ] i were not affected by acetazolamide treatment. However, AQP1 content was significantly reduced from 43.15±2.97 to 23.85±1.92. Therefore, it can be concluded that the regulation effect of acetazolamide on pH and [ Ca 2+ ] i was depended on the existence of HCO 3 , while its inhibition on AQP1 expression was not related to the change of intracellular pH and [ Ca 2+ ] i.更多还原