AQP4基因敲除小鼠PERG的变化及激光诱导其高眼压模型的建立

发布时间：2018-01-30 19:38

本文关键词： 青光眼电生理眼内压动物实验模型　出处：《南京医科大学》2012年硕士论文　论文类型：学位论文

【摘要】：目的：使用图形视网膜电流图（pattern electroretinogram, PERG）评估野生型（wide type, WT）小鼠和水通道蛋白4（aquaporin4,AQP4）基因敲除（knock out,KO）小鼠（CD1背景鼠）视网膜神经节细胞（retinal ganglion cell, RGC）的功能，逆向研究AQP4基因在维持小鼠视网膜正常生理功能中所扮演的角色。通过激光光凝角膜缘和巩膜上静脉的方法，观察AQP4在高眼压模型下对眼压（intraocular pressure, IOP）变化的影响，建立水通道蛋白4基因敲除小鼠的高眼压模型。方法：1. AQP4基因敲除小鼠和野生型小鼠各18只，2%水合氯醛0.2ml/10g腹腔注射麻醉，采用自制电极测量各小鼠的图形视网膜电流图，对结果进行统计分析。2.使用532nm二极管激光光凝角膜缘和巩膜上静脉的方法制作AQP4基因敲除小鼠和野生型小鼠的高眼压模型，使用IcareLAB回弹式眼压计（rebound tonometer, RBT）测量小鼠术前及术后的眼压值，观察AQP4基因敲除小鼠和野生型小鼠各自眼压的变化情况。结果：1. AQP4基因敲除小鼠（n=18）图性视网膜电流图的P50振幅（5.53±1.31）uV，N95振幅（7.71±1.89）uV。野生型小鼠（n=18）的P50振幅（8.14±1.24）uV，N95振幅（11.30±2.61）uV。AQP4基因敲除小鼠的P50和N95的振幅较野生型小鼠的低（P＜0.01），潜伏期也较野生型小鼠的提前。2.AQP4基因敲除小鼠（18只鼠36眼）光凝手术前平均眼压（6.61±0.90）mmHg，野生型小鼠（18只鼠36眼）光凝手术前平均眼压（7.31±0.98）mmHg，基因敲除小鼠和野生型小鼠之间的基础平均眼压值存在微小但有统计学意义的差异（P＜0.05）。光凝手术后AQP4基因敲除小鼠（n=18）和野生型小鼠（n=18）的眼压值均在术后第1d上升到最高点，达基础眼压的两倍多，KO（14.78±1.80）mmHg，WT（16.44±1.46）mmHg。之后两种小鼠的眼压值均逐渐降低，在第8d时基本降至术前基础眼压水平。在激光光凝角膜缘和巩膜上静脉的方法下，AQP4基因敲除小鼠和野生型小鼠均表现出眼压升高，，两种小鼠眼压值变化的幅度基本一致，两种小鼠之间的眼压差异始终存在并贯穿整个实验过程。结论：1.图形视网膜电流图能很好地反应小鼠视网膜神经节细胞的功能，AQP4基因缺失可能直接破坏了小鼠的RGCs功能，对小鼠的视网膜电生理功能产生了不良影响。2.采用532nm二极管激光光凝角膜缘和巩膜上静脉的方法可以使AQP4基因敲除小鼠和野生型小鼠的眼内压短时间升高，成功制作了新的青光眼动物模型。未来的研究可能通过抑制AQP4在睫状体非色素上皮细胞的表达而寻求到新的降低眼内压的方法。
[Abstract]:Objective: to evaluate wild type type by pattern electroretinogrammetry (Perg). WT-mice and aquaporin4AQP4) knockout out. The function of retinal ganglion cells (RGCs). Reverse study of the role of AQP4 gene in maintaining normal physiological function of mouse retina by laser photocoagulation of limbus cornea and superior scleral vein. To observe the effect of AQP4 on intraocular pressure (IOP) in the model of high intraocular pressure (IOP). A high IOP model of aquaporin-4 knockout mice was established. Methods AQP4 gene knockout mice and wild type mice were anesthetized by intraperitoneal injection of 2% chloral hydrate 0.2 ml / 10 g. The electroretinogram of each mouse was measured by self-made electrode. Statistical analysis of the results .2. using 532nm diode laser photocoagulation of corneal limbus and superior scleral vein to make AQP4 gene knockout mice and wild-type mice model of high intraocular pressure. Intraocular pressure (IOP) was measured before and after operation by IcareLAB rebound intraocular pressure meter (IcareLAB). To observe the change of intraocular pressure in AQP4 knockout mice and wild type mice. Results the P50 amplitude of AQP4 gene knockout mice was 5.53 卤1.31 UV. The P50 amplitude of N95 was 7.71 卤1.89uV. the P50 amplitude was 8.14 卤1.24uV in wild type mice. The amplitudes of P50 and N95 in N95 knockout mice were lower than those in wild type mice (11.30 卤2.61 渭 V.AQP4 knockout mice, P < 0.01). The incubation period was also earlier than that of wild-type mice. 2. The mean IOP before photocoagulation was 6.61 卤0.90 mm Hg in 18 mice with AQP4 knockout. The mean intraocular pressure (IOP) before photocoagulation was 7.31 卤0.98 mmHg in 18 mice (36 eyes). There was a small but statistically significant difference in the basic mean IOP between the knockout mice and the wild-type mice (P < 0.05). AQP4 gene knockout mice after photocoagulation (P < 0.05). Intraocular pressure (IOP) increased to the highest point on the 1st day after operation. The intraocular pressure (IOP) of the two mice decreased gradually after reaching the basic intraocular pressure (IOP) of 14.78 卤1.80 mm HgG and 16.44 卤1.46 mHg. On the 8th day, the intraocular pressure (IOP) was basically reduced to the preoperative basic intraocular pressure level. The intraocular pressure was increased in both mice and wild-type mice by laser photocoagulation of limbus cornea and superior scleral vein. The IOP values of the two kinds of mice were basically the same, and the IOP differences between the two kinds of mice existed all the time and ran through the whole experiment process. Conclusion the pattern electroretinogram can well reflect the function of retinal ganglion cells in mice. The deletion of AQP4 gene may directly destroy the RGCs function of mice. This method can make AQP4 gene knockout mice and wild-type mice intraocular by using 532nm diode laser photocoagulation of limbus cornea and superior scleral vein. Short pressure increases. A new glaucoma animal model was successfully established. Future studies may seek a new way to reduce intraocular pressure by inhibiting the expression of AQP4 in non-pigment epithelial cells of ciliary body.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R775;R-332

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