Objectives To investigate whether aquaporin 4 ( AQP4 ) gene can affect the activation of glial cells and cause the injury of retina of chronic high intraocular pressure mice models,and to discuss its possible mechanism.Methods Experimental study.The chronic high intraocular pressure models were established by burning the scleral venous of the right eye,which as the experiment group,and the left eye without any treatment as the control group.The intraocular pressure (IOP) was measured by rebound tonometer.Selected each of the successful model of chronic high intraocular pressure male AQP4 knockout mice ( AQP-/-) and their wild-type(WT) male mice 40,divided the two type of mice into five groups after scleral venous burn according to the time of establishing models(24 h,3 d,1 w,2 w,4 w after scleral venous burn),8 mouse in each group.And then producing the paraffin sections of mouse eye.Immunohistochemical staining methods was used to observe the expression of the glial fibrillary acid protein(GFAP) in retina glial cells,and observe the expression of the AQP4 in the retina of the WT mouse.Image was acquired under the fluorescence microscopy,The intraocular pressure was analyzed by t-test.Results After scleral venous burn (24 h,3 d,1 w,2 w,4 w),there were significant difference ( t =6.66-18.08,all P value ＜ 0.05 ) in the IOP of the AQP4-/-mice ( 11.30 ± 1.59,11.20 ± 1.15,10.60 ± 1.53,10.75 ± 1.45,10.45 ± 1.39) and WTmice (11.50 ±2.56,11.25 ± 1.65,10.75 ± 1.33,10.60 ± 1.33,10.40 ± 1.19) between the experimental groups and control groups(6.60 ± 0.94,6.35 ± 0.99,6.55 ± 0.94,6.45 ± 0.99,6.50 ± 0.94and 6.60 ± 1.05,6.50 ± 0.89,6.40 ± 1.09,6.30 ± 1.13,6.50 ± 1.05 ).Since 24 hours after the scleral venous burn,the expression of GAFP of the two type mice began to increase and reached to peak at 1 week after burning.This peak of WT mice was more obvious than that of AQP4-/-mice.The concentration of GAFP began to decrease at 2 weeks afere burning and reached to bottom at 4 weeks later.To the WT mice,the expression of AQP4 was remarkable higher in experimental group than that in control group at 1 week after the scleral venous burn.The expression of AQP4 was related to the expression of GAFP in high intraocular pressure of WT mice at 1 w,2 w,and 4 w after the scleral venous burn.Conclusions The chronic high intraocular pressure models can be established successfully by burning the scleral venous.AQP4 gene can affect the activation of the glial cells in chronic high intraocular pressure mice and lead to the injury of retina.%目的 研究慢性高眼压状态下小鼠水通道蛋白4(AQP4)基因是否通过影响胶质细胞的活化造成青光眼视网膜的损伤,并探讨其可能机制.方法 实验研究.采用小鼠巩膜外静脉烧烙法建立小鼠高眼压模型.应用同弹式眼压计测量小鼠眼压.右眼为慢性高眼压眼,左眼为对照眼.选择造模成功的慢性高眼压雄性AQP4基因敲除(AQP4-/-)小鼠及其背景雄性野生型(WT)小鼠各40只,根据慢性高眼压模型建立的时间(手术结束时开始计算),将两种小鼠分别分为5组(24h、3d、1、2、4周),每组8只.石蜡切片行AQP4-/-小鼠及WT小鼠胶质细胞中胶质纤维酸性蛋白(GFAP)和WT小鼠AQP4的免疫组织化学染色,荧光显微镜下采集图像.小鼠眼压的组间比较采用t检验.结果 小鼠巩膜外静脉烧烙术后24h、3d、1、2、4周,AQP4-/-小鼠和WT小鼠慢性高眼压眼眼压(11.30±1.59、11.20±1.15、10.60±1.53、10.75±1.45、10.45±1.39和11.50±2.56、11.25±1.65、10.75±1.33、10.60±1.33、10.40±1.19)与对照眼(6.60±0.94、6.35±0.99、6.55±0.94、6.45±0.99、6.50±0.94和6.60±1.05、6.50±0.89、6.40±1.09、6.30±1.13、6.50±1.05)相比,差异有统计学意义(t=6.66～18.08,P＜0.05).免疫荧光染色结果显示,术后24h两种小鼠慢性高眼压眼的GFAP表达量均开始增加,1周时表达水平最高,2周时开始减弱,4周时最弱,但仍高于对照眼；术后1周时WT小鼠较AQP4-/-小鼠GFAP的表达量增加更为明显；WT小鼠在眼压升高1周时AQP 4的表达量较对照眼明显增加,2、4周时仍高于对照眼；WT小鼠在眼压升高1、2、4周时AQP4表达水平与GFAP表达水平均具有一致性.结论 小鼠巩膜外静脉烧烙的方法能有效升高小鼠眼压；高眼压状态下AQP4基因可能通过影响小鼠胶质细胞的活化造成青光眼视网膜损伤.
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