本文選題：X射線 + 放射性白內(nèi)障　； 參考：《蘇州大學》2010年碩士論文

【摘要】： 目的:利用動物實驗,通過X射線照射S-D大鼠雙眼建立放射性白內(nèi)障動物模型,觀察X射線劑量與白內(nèi)障嚴重程度之間的關系;檢測受不同劑量X射線照射的大鼠晶狀體中水通道蛋白1(AQP1)的表達水平,探討水通道蛋白1與放射性白內(nèi)障形成之間的關系。 方法:照射前選取8只大鼠作為正常對照組,實驗照射組大鼠24只,平均分成3組每組8只,一次性分別照射X射線(雙眼同時照射)5、15、25 Gy.實驗對照組24只,分成3組每組8只,分別作為5、15、25 Gy的對照組行假照射(照射光線中不含X射線,其它條件分別與對應各照射組相同)。每只大鼠隨機取一只眼睛(本實驗取右眼)為研究對象。所有大鼠分別于照射后1、3、5、7、15、30、45、60、90 d,腹腔注射3.6%水合氯醛麻醉后,用雙星明滴眼液充分散瞳,裂隙燈顯微鏡觀察并記錄晶狀體混濁情況;運用免疫組織化學方法(SP法)對AQP1在大鼠晶狀體上的表達進行定位檢測,并采用Motic Med 6.0數(shù)碼醫(yī)學圖像分析系統(tǒng)半定量檢測AQP1的表達;運用逆轉(zhuǎn)錄聚合酶鏈反應(RT-PCR)及計算機圖像分析技術檢測AQP1在各組大鼠晶狀體上的表達變化;應用SPSS11.5統(tǒng)計軟件對結果進行統(tǒng)計分析,采用Independent Samples T-Test法和One-Way-ANOVA法分別對計量資料進行獨立樣本T檢驗和單因素方差分析,并使用LSD法進行組間比較。 結果: 1.組織學:觀察3個月,各實驗對照組和5 Gy組大鼠的晶狀體均未出現(xiàn)混濁。15 Gy照射組于照射后45 d晶狀體后囊和后皮質(zhì)開始出現(xiàn)細小、散在白點狀混濁,繼而融合成不均勻較密集點狀、片狀混濁,進一步發(fā)展后皮質(zhì)出現(xiàn)較均勻致密混濁,前皮質(zhì)、前囊和核部輕度混濁。到3個月時62.5%的晶狀體達Ⅱ期混濁,37.5%達Ⅲ期混濁。25 Gy照射組亦出現(xiàn)類似變化,但是晶體發(fā)生混濁的時間更早、速度更快、混濁更明顯。于照射后30d后囊即開始出現(xiàn)點狀混濁,此后病變迅速發(fā)展,到45d時即有37.5%的晶體全部混濁。到3個月的時候除1例為后囊下明顯混濁外,其余7例晶狀體均完全混濁。 2.免疫組織化學和計算機圖像分析結果顯示:AQP1在大鼠晶狀體上陽性表達的部位位于晶狀體前囊膜的晶體上皮細胞(lens epithelial cells,LECs)的胞膜,胞膜上可見程度不等的棕黃色顆粒。各組均可見到其陽性表達,隨照射量增大,各實驗組LECs上的AQP1均較空白組表達減少,差異有顯著性意義(P0.05)。各實驗組AQP1的表達隨照射劑量的增加而減少,組內(nèi)差異有統(tǒng)計學意義(P0.05)。 3.RT-PCR和計算機圖像分析結果顯示:在mRNA水平AQP1在大鼠晶狀體LECs胞膜表達,實驗組和空白組的晶狀體進行RT-PCR均可獲得AQP1的陽性目的基因,不同照射量實驗組晶狀體的AQP1的表達均較空白組有不同程度的減少,差異有統(tǒng)計學意義(P0.05)。而且隨著照射量的增加,各亞組之間AQP1的表達水平降低,變化呈劑量依賴性,差異有統(tǒng)計學意義(P0.05)。 結論:大劑量X射線在短期內(nèi)即可引起晶狀體混濁;放射性白內(nèi)障的形成速度和程度與射線劑量成正相關。在輻射線誘導的白內(nèi)障模型,大鼠晶狀體中AQP1的表達發(fā)生改變,輻射損傷可能通過減少AQP1在晶狀體上皮細胞上的表達,影響晶狀體的水代謝,以誘發(fā)白內(nèi)障在放射性白內(nèi)障中晶狀體上皮細胞內(nèi)AQP1表達減少,在白內(nèi)障形成中起重要作用。
[Abstract]:Objective: to establish a radioactive cataract animal model in the eyes of S-D rats by X ray irradiation, and to observe the relationship between the dose of X ray and the severity of cataract, and to detect the expression level of aquaporin 1 (AQP1) in the lens of rats exposed to different doses of X ray, and to explore the formation of aquaporin 1 and the formation of radioactive cataract. The relationship between them.
Methods: before irradiation, 8 rats were selected as the normal control group, and 24 rats in the experimental group were divided into 3 groups, with an average of 8 rats in each group. 24 rats were irradiated with X ray (double binocular simultaneous irradiation) 5,15,25 Gy. experimental control group, and 8 rats in each group were divided into 3 groups of 8, respectively, which were respectively irradiated with no X rays in the light rays and other conditions. Each rat was treated with one eye (the right eye) randomly. All rats were treated with 1,3,5,7,15,30,45,60,90 d after irradiation. After intraperitoneal injection of 3.6% chloral anaesthesia, double star eyedrops were used to fully diffuse pupil, slit lamp microscopes were observed and recorded in the lens opacity; immunization was used. The expression of AQP1 in rat lens was detected by histochemical method (SP method), and the expression of AQP1 was detected by Motic Med 6 digital medical image analysis system. Reverse transcription polymerase chain reaction (RT-PCR) and computer image analysis were used to detect the expression of AQP1 in the lens of rats in each group; and SPSS11. was applied. 5 statistical software was used to analyze the results. The Independent Samples T-Test method and One-Way-ANOVA method were used to carry out independent sample T test and single factor analysis of variance, and the LSD method was used to compare the data between groups.
Result:
1. histology: after 3 months of observation, the lens of the experimental control group and the 5 Gy group had no cloudy.15 Gy irradiation group, and the posterior capsule and the posterior cortex began to appear in the posterior capsule and the posterior cortex after the irradiation. In the cortex, the anterior capsule and the nucleus, 62.5% of the lens reached phase II turbidity by 3 months, and the 37.5% period of phase III cloudy.25 Gy irradiation group also appeared similar changes, but the time of turbidity in the crystal was earlier, the speed was faster, and the turbidity was more obvious. After 30d, the PACA began to appear turbid, and then the pathological changes developed rapidly, when 45d was 37.. 5% of the lenses were all cloudy. At 3 months, 7 of the 7 patients were completely opacification except for 1 cases of posterior capsular opacification.
2. the results of immuno histochemistry and computer image analysis showed that the positive expression of AQP1 in the lens of the rat was located in the membrane of the lens epithelial cells (lens epithelial cells, LECs) in the anterior capsule of the lens, and the brown yellow granules of different degree were found on the membrane. The positive expression was seen in each group, with the increase of the irradiation amount, the experimental group was LE. The expression of AQP1 on Cs was less than that in the blank group, and the difference was significant (P0.05). The expression of AQP1 in the experimental group decreased with the increase of irradiation dose, and the difference in the group was statistically significant (P0.05).
The results of 3.RT-PCR and computer image analysis showed that the expression of LECs cell membrane in the rat lens at the level of AQP1 at mRNA level, the positive target gene of AQP1 in the experimental group and the blank group could be obtained by RT-PCR, and the expression of AQP1 in the lens of the experimental group of different irradiated groups were all lower than that in the blank group, and the difference was statistically significant (P0.05). Moreover, with the increase of irradiation dose, the expression level of AQP1 in all subgroups decreased, and the change was dose-dependent, and the difference was statistically significant (P0.05).
Conclusion: large dose X rays can cause lens opacities in a short period of time. The velocity and degree of radiation cataract formation is positively correlated with the dose of ray. In the cataract model induced by radiation, the expression of AQP1 in the lens of rat is changed. The radiation damage may affect the crystalline form by reducing the expression of AQP1 on the lens epithelial cells and affecting the crystalline form. The water metabolism of the body can induce the decrease of AQP1 expression in MICROTEK epithelial cells, and play an important role in the formation of cataract.
【學位授予單位】：蘇州大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R776.1

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