不同人工泪液对兔角膜上皮修复作用的研究

发布时间：2018-05-18 23:04

本文选题：兔子 + 人工泪液　；参考：《浙江大学》2017年硕士论文

【摘要】：研究背景:人角膜由上皮层、前弹力层、基质层、后弹力层及内皮层五部分组成,角膜各层结构完整,功能和代谢正常,共同维持着角膜的整体透明性。角膜上皮细胞层处于角膜的最外层,对于维持角膜正常形态结构及功能,抵抗外力侵袭起着十分重要的作用。这归功于上皮细胞之间,上皮细胞与基底膜之间的连接,以及上皮细胞的持续自我更新。上皮细胞受到损害、消亡,进而影响细胞之间,及细胞与基底膜之间的连接,导致上皮层结构功能的不稳定性,使角膜组织容易受到外界致病因子的侵害。世界卫生组织声明,角膜损伤仅次于角膜感染,成为第二大潜在致盲原因。某些药物长期全身、局部使用,外伤,感染,手术等均可引起角膜上皮的损伤。不完全、不及时的角膜上皮修复容易引起角膜感染、角膜新生血管长入、溃疡、穿孔、角膜白斑、斑翳甚至失明。通常情况下,角膜上皮缺损,特别是小范围损伤,一般自我修复较快,但某些损伤,可导致持续性角膜上皮缺损,而相对严重的角膜上皮损伤需要尽快得到治疗,以促进角膜上皮的再生修复及避免角膜感染及斑翳。角膜上皮细胞之间紧密连接的形成,在角膜上皮修复过程中是非常重要的一步,可以促进上皮屏障的完整形成,抵御病原微生物的入侵,防止角膜溃疡甚至穿孔。近几年,促进角膜上皮快速修复得到临床越来越多的重视,尽管有很多滴眼液被认为可以促进角膜上皮修复,但由于角膜上皮细胞具有较强的自我修复能力,故很难明确不同眼药水在角膜上皮修复中的差别。临床上,人工泪液被广泛应用于促进角膜上皮修复,可是选择何种人工泪液一直没有相应的共识。搜索文献发现,关于人工泪液对角膜上皮修复是否存在促进作用的相关临床报道及研究较少。人工泪液能够缓解眼部干涩,改善泪膜功能,延长泪膜破裂时间,学者们已经研究透彻,但是,人工泪液除了能改善泪膜功能,对于角膜损伤修复,究竟有没有作用尚不得而知。本研究拟探讨不同人工泪液对角膜上皮修复的影响,首先建立兔角膜损伤模型,模拟人类角膜损伤,观察不同人工泪液对兔角膜上皮再生修复是否存在促进作用,同时比较不同人工泪液时间是否存在差异,并在角膜上皮修复不同阶段行病理学及扫描电镜检查,以期增加对人工泪液影响角膜上皮修复的认识,从而为临床用药提供依据。研究目的:观察不同人工泪液对兔角膜上皮的修复过程,评估人工泪液是否存在促进角膜上皮修复作用,及不同人工泪液之间是否存在差异。研究方法:选取相同年龄的35只(35眼)新西兰长耳兔,建立角膜上皮损伤模型(6mm直径圆形中央上皮缺损),随机将动物模型分为7组,分别为实验组:0.1%玻璃酸钠滴眼液组,0.3%玻璃酸钠滴眼液组,羟糖苷滴眼液组,聚乙二醇滴眼液组,0.5%羧甲基纤维素钠滴眼液组,生理盐水组及空白对照组。实验组均给予相应滴眼液,使用频率为1滴/次,4次/日。并且在损伤模型建立之后0小时,24小时,36小时,48小时,60小时及72小时,进行裂隙灯观察,拍照,并在不同时期,进行病理学检查上皮细胞与基质连接及上皮细胞排列情况,并扫描电镜观察角膜上皮超微结构。比较不同实验组之间的差异,及实验组与对照组之间的差异。所有数据均采用SPSS 17.0软件包进行统计学处理,采用Tamhane'sT2进行实验组间两两对比分析,运用独立样本T检验进行实验组与对照组对比分析。P0.05为差异有统计学意义。(以a=0.05为检验水准)。结果:1.所有损伤角膜上皮(包括实验组及对照组)均在72小时以内修复完全(形态学上),大部分损伤眼角膜上皮在60小时以内修复完全(形态学上),少部分在48小时左右修复完全。2.72小时,对实验组兔眼角膜做病理组织学检查发现:所有再生的角膜上皮细胞层数完整,排列整齐,与基底部贴合紧密。扫面电镜观察超微结构时发现,24小时,0.3%玻璃酸钠滴眼液组角膜上皮缺损区仍有基底膜暴露,生理盐水滴眼液组基底膜已被再生角膜上皮覆盖,72小时,生理盐水组角膜上皮细胞基质内含有大量液泡,而0.3%玻璃酸钠滴眼液组中几乎没有,与正常角膜上皮形态无异。3.空白对照组角膜上皮再生修复所需时间明显比实验组长,对比有统计学意义。实验组之间两两比较,差异无统计学意义。玻璃酸钠对角膜上皮的修复作用于浓度有关。结论:1.人工泪液对兔角膜上皮损伤均有修复作用。2.玻璃酸钠滴眼液对兔角膜上皮的修复能力与其浓度有关。3.生理盐水可促进兔角膜上皮细胞较快的再生、移行及定植,但72小时修复再生的上皮细胞与正常角膜上皮细胞内微观结构有差异。
[Abstract]:Background: human cornea consists of five parts of the epithelium, the front elastic layer, the matrix layer, the posterior elastic layer and the endothelial layer. All the layers of the cornea are intact, function and metabolize normally to maintain the overall transparency of the cornea. The corneal epithelial cell layer is in the outer layer of the cornea, to maintain the normal structure and function of the cornea and resist external force invasion. It is of great importance. This is attributed to the connection between epithelial cells, epithelial cells and the basement membrane, and the continuous self renewal of epithelial cells. The epithelial cells are damaged and disappeared, and the connections between the cells and the basement membrane are affected, which lead to the instability of the structure and function of the epithelium and make the cornea tissue easy to suffer. The WHO declared that corneal injury is the second most potential cause of blindness. Some drugs can cause corneal epithelial damage for long time, local use, trauma, infection, and surgery. Incomplete corneal epithelial repair is easy to cause corneal infection and corneal neoplasma. Canes, perforations, perforations, corneal leukoplakia, pannus and even blindness. Generally, corneal epithelial defects, especially small range injuries, are usually repaired faster, but some injuries can lead to persistent corneal epithelial defect, and a relatively severe corneal epithelial injury needs to be treated as soon as possible to promote regenerative repair and avoidance of corneal epithelium. The formation of close connection between corneal epithelial cells is a very important step in the process of corneal epithelial repair. It can promote the complete formation of the epithelial barrier, resist the invasion of the pathogenic microorganism, prevent corneal ulcer and even perforate. In recent years, the rapid repair of corneal epithelium has gained more and more weight in clinic. Although many eye drops are considered to promote corneal epithelial repair, it is difficult to identify the difference in corneal epithelial repair due to the strong self repair ability of corneal epithelial cells. There is a corresponding consensus. The search literature has found that there are few clinical reports and studies about the effect of artificial tears on the promotion of corneal epithelial repair. Artificial tears can relieve eye dryness, improve tear film function, and prolong tear film rupture time. Scholars have studied thoroughly, but artificial tears can improve tear film function, but This study is to explore the effects of different artificial tears on corneal epithelial repair. First of all, a rabbit corneal injury model is established to simulate human corneal injury, and the effect of different artificial tears on corneal epithelial regeneration and repair in rabbits is observed and the different artificial tears are compared. In order to increase the understanding of the influence of artificial tears on corneal epithelium repair and provide a basis for clinical use, the purpose of this study is to observe the repair process of rabbit corneal epithelium with different artificial tears and to evaluate the existence of the promotion angle of artificial tears. Study methods: 35 New Zealand long ear rabbits (35 eyes) of the same age were selected to establish corneal epithelial injury model (6mm diameter circular central epithelial defect), and the animal models were randomly divided into 7 groups, 0.1% Sodium Hyaluronate Eye Drops group and 0.3% sodium hyaluronate drop. Eye fluid group, hydroxy glucoside eye drop group, Polyethylene Glycol Eye Drops group, 0.5% Carboxymethylcellulose Sodium Eye Drops group, saline group and blank control group. The experimental group was given the corresponding eye drops, using 1 drops / times, 4 times / day. And the slit lamp was carried out at 0 hours, 24 hours, 36 hours, 48 hours, 60 hours and 72 hours after the damage model was established. Observation, photographing, and at different periods, pathological examination of epithelial cells and matrix connection and the arrangement of epithelial cells, scanning electron microscope to observe the ultrastructure of corneal epithelium. Compare the differences between the different experimental groups and the differences between the experimental group and the control group. All the data were statistically treated by SPSS 17 software package. 22 contrast analysis between the experimental groups was carried out, and the experimental group was compared with the control group by the independent sample T test. The difference between the experimental group and the control group was statistically significant. (a=0.05 as the test level). Results: 1. all damaged corneal epithelium (including the experimental group and the control group) were completely repaired within 72 hours (morphological) and most of the injuries were damaged within 72 hours. The corneal epithelium was repaired within 60 hours completely (morphologically), less part of the corneal epithelium was repaired for about 48 hours for complete.2.72 hours. Histopathological examination of the cornea of the rabbits in the experimental group showed that all the regenerated corneal epithelial cells were complete, arranged and fitted closely with the basal part. The ultrastructure of the corneal epithelium was found to be 24 hours, 0.. 3% in the corneal epithelial defect area of the Sodium Hyaluronate Eye Drops group, the basement membrane was still exposed, and the basement membrane of the saline eye drops was covered by the regenerated corneal epithelium. 72 hours, there were a large number of vacuoles in the matrix of the corneal epithelial cells in the saline group, while the 0.3% Sodium Hyaluronate Eye Drops group had almost no one in the normal corneal epithelium, which was no different from the normal corneal epithelium in.3. blank control. The time required for regenerative repair of corneal epithelium was significantly longer than that of the experimental group, and the comparison was statistically significant. There was no significant difference between the 22 experimental groups. The repair effect of sodium hyaluronate on corneal epithelium was related to the concentration. Conclusion: 1. artificial tears have a repair effect on corneal epithelial injury in rabbits with.2. Sodium Hyaluronate Eye Drops on the rabbit cornea The repair ability of skin and its concentration related to.3. physiological saline can promote the rapid regeneration, migration and colonization of rabbit corneal epithelial cells, but the microstructure of the epithelial cells regenerated by 72 hours is different from that in the normal corneal epithelial cells.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R77

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