阿瓦斯汀抑制兔眼碱烧伤角膜新生血管的研究

发布时间：2018-10-19 11:46

【摘要】：目的：通过研究阿瓦斯汀(Avastin)结膜下给药对兔眼碱烧伤角膜新生血管的影响,探索阿瓦斯汀治疗角膜新生血管的安全性及其有效性,为临床应用阿瓦斯汀治疗角膜新生血管性疾病提供实验依据。方法：选用成年新西兰雄性大白兔60只(苏州大学动物实验中心),体重在2-3Kg之间,健康状况良好,均取左眼作为实验眼,随机抽选45只兔左眼用1mol/LNaOH溶液烧伤,建立碱烧伤角膜新生血管模型。未造模的15只兔左眼设为A正常对照组,成功造模的45只兔再随机分为三组,每组15只,分别为B模型对照组(造模成功后不做治疗),C新药干预组(术后2小时结膜下注射2.5mg/0.1ml阿瓦斯汀),D常规治疗照组(术后2小时结膜下注射地塞米松0.5mg/0.1ml),于碱烧伤后4、7、14天拍照记录,观察活体兔眼结膜充血、角膜水肿浑浊、上皮的完整性及新生血管的生长情况,定量计算角膜新生血管面积,并于三个时间点每组随机取5只兔用空气栓塞法处死,取实验眼角膜行病理检查及免疫组化检测角膜VEGF和CD31的表达。结果： 1、碱烧伤4天,造模组所有兔眼均出现明显的炎症反应,结膜充血水肿,角膜水肿浑浊,上皮不完整,B组角膜缘见少许新生血管长入,C、D组未发现有角膜新生血管,7天,造模组所有兔眼结膜充血明显,角膜新生血管生长旺盛,自角膜缘呈毛刷样向烧伤区爬行,10-14点区域新生血管出现最多。14天,角、结膜充血水肿减轻,角膜上皮修复,角膜浑浊,中央烧伤区最为致密,角膜新生血管有所消退。 2、病理显示,正常对照组角膜上皮完整,层次清晰,角膜基质纤维排列整齐,无炎细胞及血管,术后4天造模组角膜上皮连续性中断,上皮下及浅基质层见少量新生血管芽,管腔少量血细胞,大量炎细胞主要聚集于角膜浅层,7天时角膜新生血管密集,并向深基质层长入,管腔粗大且充满血细胞。14天时,炎细胞减少,表面新生血管部分消退,基质层见管径粗大的血管,血细胞充盈,在不同时间点,C、D组之间差别无统计学差异,C、D组与B组之间比较有统计学差异。 3、角膜VEGF检测结果：碱烧伤4天,造模组所有兔眼角膜均出现VEGF,7天VEGF表现强阳性,14天时密度降低,三个时间点统计,C、D组之间、/EGF表达强度无统计学差异,C、D组VEGF表达强度较B组轻,与B组之间比较有统计学差异。 4、角膜CD31检测结果示：CD31在正常兔眼角膜组织中无表达,造模组兔角膜中均有表达,7天时表达强阳性,各时间点C、D治疗组阳性细胞数明显低于模型对照组,差异有统计学意义。结论： 1、兔眼角膜碱烧伤4天角膜开始有新生血管长入,烧伤后7天时结膜炎症反应比较重,角膜水肿浑浊,新生血管生长旺盛,14天结膜炎症反应减轻,角膜新生血管有所消退。 2、阿瓦斯汀与地塞米松对兔眼碱烧伤后形成的新生血管都有很好的抑制作用,与B造模组相比,C、D组在三个不同时间点检测到的新生血管、VEGF及CD31均显著降低,C组与D组在三个不同的时间点检测的各项指标无显著差异。 3、阿瓦斯汀治疗兔眼角膜新生血管简单、安全、效果显著,在观察期内未见并发症发生。
[Abstract]:Purpose: Objective To explore the safety and efficacy of Avastin in the treatment of corneal neovascularization by studying the effect of Avastin on corneal neovascularization in rabbits, and to provide an experiment for the clinical application of Avastin in the treatment of corneal neovascularization. Basis. Methods: 60 adult New Zealand white rabbits were selected (Suzhou University Animal Experiment Center), the body weight was 2-3Kg, the health condition was good, the left eye was taken as the experimental eye, 45 rabbits were randomly selected to burn the left eye with 1mol/ L NaOH solution, and alkali burn was established. The corneal neovascularization model was established. 15 rabbits were randomly divided into three groups, 15 rabbits in each group were randomly divided into three groups, 15 rabbits in each group were respectively B model control group (not treated after the model was successful), and the C drug intervention group (2 hours after operation) was injected with 2. 5mg/ 0. 1 ml of Avastin, D conventional treatment group (dexamethasone 0.5 mg/ 0. 1ml at 2 hours after operation), 4, 7 and 14 days after alkali burn, observation of conjunctival congestion, corneal edema, integrity of epithelium and growth of newborn blood vessels were observed, and quantitative calculation was carried out. The corneal neovascularization area was randomly taken at three time points, 5 rabbits were killed by air embolism method, the pathological examination and immunohistochemistry were used to detect corneal VEGF. 鍜孋D Results: 1. After 4 days of alkali burn, there were obvious inflammatory response, conjunctival congestion edema, corneal edema and incomplete epithelium in all rabbit eyes. A few new blood vessels were seen in group B limbal, C, D. The corneal neovascularization was not found in the group, 7 days, the conjunctival congestion of all rabbits in the building module was obvious, the corneal neovascularization was vigorous, the brush-like vein appeared on the limbal edge to crawl into the burn area, and the newly-formed blood vessel in the area of 10-14 points appeared the most. 14 days, the angle, conjunctival hyperemia and edema were alleviated, and the corneal epithelium was repaired. Corneal opacity, central burn area most In the normal control group, the corneal epithelium is intact, the level is clear, the corneal stroma fibers are arranged in order, there is no inflammatory cells and blood vessels, the continuity of corneal epithelium is interrupted, the upper subcutaneous layer and the shallow matrix are formed on the 4-day post-operation. The layer is seen in a small amount of fresh blood vessel bud, small amount of blood cells in the tube cavity, a large number of inflammatory cells are mainly concentrated on the superficial layer of the cornea, the corneal neovascularization is dense at 7 days, and the deep matrix layer is long inserted, the tube cavity is large and full of blood cells. 14 days, the inflammatory cells are reduced, the surface fresh blood vessel part is resolved, There was no statistical difference between C and D groups at different time points. There was a statistically significant difference between group C, group D and group B. 3. The results of corneal VEGF detection: 4 days after alkali burn, VEGF was observed in all eyes of all rabbits, 7 days VEGF expression was strongly positive, the density was decreased at 14 days, and there was no statistical difference between the three time points statistics, C and D groups and/ EGF expression intensity. The expression of VEGF in group C and D The results showed that CD31 was not expressed in the cornea of normal rabbit eyes, and in the cornea of the module rabbits, the expression was strongly positive at 7 days, and the C and D treatment groups at each time point were positive. positive fine The number of cells was significantly lower than that in the model control group, and the difference was statistically significant. Conclusion: 1. The cornea of the cornea in the cornea of rabbit cornea began to grow in 4 days, the conjunctivitis was heavier in 7 days after burn, and the corneal edema was cloudy and new. In addition, Avastin and dexamethasone have a good inhibitory effect on new blood vessels formed after ocular alkali burn in rabbits, and C, D compared with B-building modules. Neovascular, VEGF and CD31 detected at three different time points There was no significant difference between group C and group D at three different time points.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R965

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