中波紫外线诱导SD大鼠白内障模型的实验研究

发布时间：2018-04-23 20:45

本文选题：中波紫外线 + 晶状体上皮细胞　；参考：《泸州医学院》2010年硕士论文

【摘要】： 目的：探讨用中波紫外线照射SD大鼠建立紫外线诱导的动物白内障模型及对比三种晶状体图像采集的效果,从而为进一步研究白内障的形成,混浊程度的评定及白内障的防治提供相关实验依据。方法：1.选用3月龄的一级大白鼠20只,随机分为实验组和对照组,实验组大鼠置于20×40×20cm3的笼中,笼子上方安置中波紫外线光源,光源距离大鼠约40cm,给予每天2小时中波紫外线照射；对照组不给于任何干预处理。选择不同照射时间段大鼠(0天、5天、20天、40天)摘取眼球去除角膜后在眼前节照相系统下采集晶状体混浊程度及晶状体核的颜色变化。然后环形剪下晶状体前囊膜,细胞面向上铺于玻片上,TUNEL法检测晶状体上皮细胞凋亡情况。观察晶状体上皮细胞的形态变化。2.另选4只大白鼠,用中波紫外线照射40天,采集眼球后对8只眼球同时选用三种方法采集白内障晶状体图像,并对比分析三种方法所获图像的效果。结果：1.眼前节照相系统观察晶状体：实验组结果显示经中波紫外线照射活体SD大鼠每天2小时,在第5天时晶状体保持透明、无明显混浊,随着照射时间的延长,在20天时出现皮质轻度混浊、多集中在“Y”字缝附近,呈现前部片状、羽毛状朝向赤道部的浅层皮质混浊,颜色呈白色,未见晶状体核的透明性及颜色改变。在40天时前囊下皮质混浊程度更加明显,前部浅皮质层及深皮质层均可观察到白色混浊,范围可以扩大到近赤道部；在手术摘取前囊膜时可见多数大鼠浅层混浊的皮质与前囊膜粘连较正常组紧密。40天照射组的晶状体核未发现有明显颜色变化。对照组未发现晶状体皮质及晶状体核有明显混浊和颜色的变化,仍保持其透明性。2. TUNEL细胞凋亡检测：DNA缺口末端原位检测部分大鼠晶状体上皮细胞发现经紫外线照射活体大鼠5天时即可发现晶状体前囊膜上皮细胞出现凋亡,凋亡的细胞呈棕黄色。3.细胞形态变化：5天组的细胞形态、大小与对照组相比无明显改变。40天组部分晶状体上皮细胞出现老化现象,可见大量的晶状体纤维与晶状体上皮细胞相连。4.对同一只白内障晶状体选用不同的三种晶状体采集方法获得晶状体图像,相比而言本实验设计的晶状体托板联合眼前节照相系统法采集的图像较其它两种方法获得的图像更清晰,结果稳定。结论：1.本实验方法成功实现了紫外线辐射性白内障大鼠模型的建立。2.中波紫外线是导致白内障发生的原因之一。3.晶状体混浊程度与中波紫外线照射的时间长短呈正相关。4.中波紫外线照射活体SD大鼠眼可以造成晶状体前囊膜上皮细胞凋亡。5.三种白内障晶状体图像采集方法各有优缺点,本实验采用黑色晶状体托板联合眼前节照相系统可以获得理想的晶状体图像,与其它两种方法相比较具有明显优势。
[Abstract]:Objective: to investigate the effects of ultraviolet irradiation on the formation of cataract in SD rats, and to compare the effects of three kinds of lens image collection in order to further study the formation of cataract. The evaluation of turbidity degree and the prevention and treatment of cataract provide relevant experimental basis. Method 1: 1. Twenty 3-month-old first-grade rats were randomly divided into experimental group and control group. Rats in the experimental group were placed in a cage of 20 脳 40 脳 20cm3, and ultraviolet B light source was placed above the cage. The distance from the light source to the rats was about 40 cm, and the rats were irradiated with ultraviolet B for 2 hours a day. The control group was not given any intervention. The degree of lens opacity and the color change of lens nucleus were collected under the anterior segment radiography system after the eyeball was removed from the cornea. Then the lens anterior capsule membrane was cut in a circular way, and the apoptosis of lens epithelial cells was detected by Tunel method. To observe the morphologic changes of lens epithelial cells. Another 4 rats were irradiated with ultraviolet B for 40 days. The lens images of 8 eyes were collected simultaneously by three methods, and the results of the three methods were compared and analyzed. The result is 1: 1. The results of experimental group showed that SD rats were exposed to UVB for 2 hours a day. On the 5th day, the lens remained transparent and had no obvious opacity, with the prolongation of irradiation time. On the 20th day, the cortex was slightly turbid, mostly located near the "Y" syringe, showing the anterior flake, feathery towards the equatorial part of the superficial cortical opacity, the color was white, and there was no change in the transparency and color of the lens nucleus. At 40 days, the degree of anterior subcapsular cortex opacity was more obvious, and white turbidity could be observed in the anterior superficial cortical layer and deep cortical layer, and the range could be extended to the near equatorial part. When the anterior capsule membrane was removed, the adhesion between the superficial cortex and the anterior capsule membrane of most of the rats was closer than that of the normal group. No significant color changes were found in the lens nucleus of the irradiation group after 40 days of irradiation. In the control group, there was no obvious turbidity and color change in the lens cortex and nucleus, and the transparency was maintained at the same time. Apoptosis of lens epithelial cells in vivo exposed to UV irradiation for 5 days, apoptosis of lens anterior capsule epithelial cells was observed, and apoptotic cells were found to be brown. 3. Morphological changes: the size of lens epithelial cells in the group of: 5 days was not significantly changed compared with that in the control group. A large number of lens fibers were connected with lens epithelial cells in the group of 40 days. Three different methods of lens acquisition were used to obtain the lens image of the same cataract lens. Compared with the other two methods, the image collected by the lens support plate combined with the anterior segment photography system in this experiment is clearer and more stable than that obtained by the other two methods. Conclusion 1. This method successfully realized the establishment of ultraviolet radiation induced cataract rat model. 2. 2. Ultraviolet B is one of the causes of cataract. The degree of lens opacity was positively correlated with the duration of ultraviolet B irradiation. Ultraviolet B irradiation induced apoptosis of lens anterior capsule epithelial cells in SD rats. The three methods of lens image acquisition have their own advantages and disadvantages. In this experiment, an ideal lens image can be obtained by using black lens plate combined with anterior segment photography system, which has obvious advantages compared with other two methods.
【学位授予单位】：泸州医学院
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R776.1

【参考文献】