水通道蛋白-1在高糖培养人晶状体上皮细胞中的表达

目的研究水通道蛋白-1（AQP-1）在糖尿病性白内障发病机制中的作用。方法从健康的供体眼中分离出人晶状体囊膜,用组织块培养法将人晶状体囊膜分别置于1g/L葡萄糖＋体积分数15%胎牛血清的DMEM培养液中（对照组）或4.5g/L葡萄糖＋15%胎牛血清的DMEM培养液中进行培养（高糖组）。培养3d、28d后观察2组人晶状体上皮细胞（LECs）的生长状态。采用免疫荧光技术和流式细胞术检测AQP-1在人LECs中表达的平均荧光强度,应用流式细胞术检测2组人LECs的凋亡和坏死情况,并进行比较。结果培养第3天2组LECs的生长形态近似,但培养28d高糖组坏死细胞增加。培养3d后高糖组人LECs的AQP-1免疫荧光表达较对照组明显增强,2组间平均荧光强度的差异有统计学意义（t=3.934,P=0.004）,但2组间LECs的凋亡率和坏死率差异均无统计学意义（t=1.681,P=0.131;t=1.064,P=0.318）。培养28d后高糖组人LECs的AQP-1免疫荧光表达较对照组弱,差异有统计学意义（t=3.069,P=0.015）,LECs凋亡率、坏死率较对照组升高,差异均有统计学意义（t=11.213,P〈0.01;t=15.778,P〈0.01）。结论 AQP-1的异常表达在糖尿病性白内障的发生发展中起重要作用。

Expression of aquaporin-1 in human lens epithelial cells by hyperosmotic stimuli

Background Diabetic cataract is a severe blinding eye disease.It is the most common complication of diabetes.Aquaporin （AQP） plays an important role for maintaining the dehydration and transparency of lens epithelial cells.Objective The aim of this study was to investigate the role of AQP-1 in the pathogenesis of diabetic cataract.Methods Human lens epithelial cells （LECs） were isolated from the healthy donor eyes and were cultured in 1 g/L glucose＋DMEM containing 150 mL/L fetal bovine serum（control group） or 4.5 g/L glucose＋DMEM containing 150 mL/L fetal bovine serum （high glucose group） for 3 days or 28 days respectively.The expression of AQP-1 in human LECs was detected using immunofluorescence techniques and flow cytometry.The apoptosis and necrosis rates of LECs were detected using flow cytometry.This study was approved by the Ethic Committee of this hospital.Results The morphology of cultured LECs was similar in the third day in control group and high glucose group.However,the slender cells were obvious increased in high glucose group compared to control group.The mean fluorescence intensity of AQP-1 was enhanced in high glucose group in comparison with control group （t=3.934,P=0.004）,but no significant differences were found in the apoptosis rate and necrosis rate between two groups （t=1.681,P=0.131;t=1.064,P=0.318） in the third day of culture.The expression of AQP-1 in the high glucose cultured group was stronger than that of the control group 3 days after hyperosmotic stimuli.The difference was significantly different（t=3.934,P=0.004）.In 28 days after culture,the fluorescence intensity of AQP-1 was obviously weakened in high glucose group compared with control group （t=3.069,P=0.015）,and the apoptosis rate and necrosis rate of LECs in high glucose group were higher than those of the control group （apoptosis rate：t=11.213,P〈0.01;necrosis rate：t=15.778,P〈0.01）.Conclusion AQP-1 may play an important role in the pathogenesis of diabetic cataract.