摘要
目的观察体外培养的人眼晶状体上皮细胞(LECs)水通道蛋白-1(AQP-1)的表达;研究地塞米松对其表达水平的影响。方法运用免疫组织化学法测定人眼LECs AQP-1的表达;用含0、1×10-8、5×10-8、10×10-8、50×10-8mol/L地塞米松的DMEM培养液培养人眼LECs7d,采用RT-PCR检测地塞米松对LECs AQP-1表达的影响。结果免疫组织化学检测示体外培养的正常人眼LECs胞膜和胞浆中可见AQP-1的表达;RT-PCR示正常人眼LECs AQP-1/β-actin吸光度比值为1·43±0·12;4种浓度地塞米松处理7d后其比值分别为1·32±0·09、1·27±0·08、1·01±0·09、0·67±0·10,当地塞米松浓度≥5×10-8mol/L时有抑制作用(P<0·05),且抑制作用随地塞米松浓度的增加逐渐增强。结论体外培养的人眼LECs中可见AQP-1的表达,AQP-1可能参与维持晶状体的脱水状态及其透明性;地塞米松抑制体外培养的人眼LECs AQP-1的表达,AQP-1可能在皮质类固醇性白内障的发生发展中起重要作用。
Objective Aquaporin-1 （AQP-1） plays an important role in maintaining liquid balance and water transportation,and it was shown to exist in lens epithelium by in vivo study. This study was to investigate the expression of AQP-1 in human lens epithelial cells （LECs） cultured in vitro and the regulation of dexamethasone. Methods Human LECs were cultured in DMEM medium containing 0,1,5,10,50×10^-8 moL/L of dexamethasone for 7 days. Immunohistochemical technique was used to measure the expression of AQP-1 in human LECs cultured in vitro. The effect of dexamethasone on the expression of AQP-1 was detected by RT-PCR semi-quantitative technique. Results LECs showed yellow-brown staining in the membrane and cytoplasm, indicating a positive expression of （ AQP-1 ）. The absorption scale value of AQP-1/β-actin in cultured LECs without dexamethasone was 1.43±0. 12,while 1.32±0. 09,1.27±0.08,1.01±0.09,0. 67±0. 10 in the groups treated with 1,5, 10, 50×10^-8 mol/L dexamethasone. The expression level of AQP-1 was gradually reduced with concentrations of dexamethasone equal to or higher than 5 ×10^-8 moL/L, showing a significant inhibition of dexamethasone on the expression of AQP-1 （P 〈 0.05）. Conclusion AQP-1 was expressed in cultured human LECs,indicating that AQP-1 may involve in the relative dehydration status and transparency of lens. The inhibition of dexamethasone on the expression of AQP-1 in LECs may be partially linked to the mechanism of glucocorticoid induced cataract （GIC）.
出处
《眼科研究》
CAS
CSCD
北大核心
2006年第5期472-475,共4页
Chinese Ophthalmic Research
基金
国家教育部基金资助(1999363)
