本文關鍵詞： 青光眼 電生理 眼內(nèi)壓 動物實驗模型　出處：《南京醫(yī)科大學》2012年碩士論文　論文類型：學位論文

【摘要】：目的：使用圖形視網(wǎng)膜電流圖（pattern electroretinogram, PERG）評估野生型（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）視網(wǎng)膜神經(jīng)節(jié)細胞（retinal ganglion cell, RGC）的功能，逆向研究AQP4基因在維持小鼠視網(wǎng)膜正常生理功能中所扮演的角色。通過激光光凝角膜緣和鞏膜上靜脈的方法，觀察AQP4在高眼壓模型下對眼壓（intraocular pressure, IOP）變化的影響，建立水通道蛋白4基因敲除小鼠的高眼壓模型。 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，采用自制電極測量各小鼠的圖形視網(wǎng)膜電流圖，對結果進行統(tǒng)計分析。2.使用532nm二極管激光光凝角膜緣和鞏膜上靜脈的方法制作AQP4基因敲除小鼠和野生型小鼠的高眼壓模型，使用IcareLAB回彈式眼壓計（rebound tonometer, RBT）測量小鼠術前及術后的眼壓值，觀察AQP4基因敲除小鼠和野生型小鼠各自眼壓的變化情況。 結果：1. AQP4基因敲除小鼠（n=18）圖性視網(wǎng)膜電流圖的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的振幅較野生型小鼠的低（P＜0.01），潛伏期也較野生型小鼠的提前。2.AQP4基因敲除小鼠（18只鼠36眼）光凝手術前平均眼壓（6.61±0.90）mmHg，野生型小鼠（18只鼠36眼）光凝手術前平均眼壓（7.31±0.98）mmHg，基因敲除小鼠和野生型小鼠之間的基礎平均眼壓值存在微小但有統(tǒng)計學意義的差異（P＜0.05）。光凝手術后AQP4基因敲除小鼠（n=18）和野生型小鼠（n=18）的眼壓值均在術后第1d上升到最高點，達基礎眼壓的兩倍多，KO（14.78±1.80）mmHg，WT（16.44±1.46）mmHg。之后兩種小鼠的眼壓值均逐漸降低，在第8d時基本降至術前基礎眼壓水平。在激光光凝角膜緣和鞏膜上靜脈的方法下，AQP4基因敲除小鼠和野生型小鼠均表現(xiàn)出眼壓升高，，兩種小鼠眼壓值變化的幅度基本一致，兩種小鼠之間的眼壓差異始終存在并貫穿整個實驗過程。 結論：1.圖形視網(wǎng)膜電流圖能很好地反應小鼠視網(wǎng)膜神經(jīng)節(jié)細胞的功能，AQP4基因缺失可能直接破壞了小鼠的RGCs功能，對小鼠的視網(wǎng)膜電生理功能產(chǎn)生了不良影響。2.采用532nm二極管激光光凝角膜緣和鞏膜上靜脈的方法可以使AQP4基因敲除小鼠和野生型小鼠的眼內(nèi)壓短時間升高，成功制作了新的青光眼動物模型。未來的研究可能通過抑制AQP4在睫狀體非色素上皮細胞的表達而尋求到新的降低眼內(nèi)壓的方法。
[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 鹵1.31 UV. The P50 amplitude of N95 was 7.71 鹵1.89uV. the P50 amplitude was 8.14 鹵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 鹵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 鹵0.90 mm Hg in 18 mice with AQP4 knockout. The mean intraocular pressure (IOP) before photocoagulation was 7.31 鹵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 鹵1.80 mm HgG and 16.44 鹵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.
【學位授予單位】：南京醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R775;R-332

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相關期刊論文 前2條

1 王曉蕾;張秀蘭;;青光眼動物模型研究進展[J];實驗動物科學;2010年01期

2 環(huán)夢佳;袁志蘭;袁松濤;錢朝旭;羅莎莎;;iCare眼壓計測量小鼠眼壓值與真實眼壓的相關性研究[J];現(xiàn)代生物醫(yī)學進展;2011年16期

本文編號：1477070