贝伐单抗抑制大鼠角膜碱烧伤后早期新生血管形成的实验研究

发布时间：2018-11-09 19:19

【摘要】： 目的：探讨贝伐单抗对大鼠角膜碱烧伤后早期新生血管形成的抑制作用。方法：28只雌性Wistar大鼠,采用1 mol/L NaOH制作大鼠右眼单眼角膜碱烧伤诱导角膜新生血管模型,随机分为实验组和对照组,每组各14只。对照组应用0.25%氯霉素滴眼液滴眼,实验组应用25mg/ml贝伐单抗+0.25%氯霉素滴眼液滴眼,均为每日两次,共连续14天。分别应用裂隙灯显微镜观察各组角膜新生血管形成状况,显微镜下记录新生血管数,HE染色,免疫组织化学法检测角膜组织中VEGF蛋白表达这四种方法,观察其对角膜新生血管的抑制作用。结果：1.裂隙灯显微镜下观察结果：对照组大鼠角膜碱烧伤后的第3天,角巩膜缘处可见毛刷状新生血管的生长；第5天角巩膜缘处新生血管侵入角膜；第7天角膜新生血管交织成网状,部分血管已侵入碱烧伤斑；第14天角膜新生血管已呈垂柳状全部侵入碱烧伤斑几乎占据整个角膜。治疗组烧伤后第5天,部分象限角膜缘可见新生血管芽；第7天,角膜新生血管侵入角膜但分布稀疏而且细小；第14天,新生血管稀疏分布在碱烧伤斑外侧。 2.显微镜下新生血管计数：对照组新生血管个数(个／400倍视野)为(137.74±21.53)、实验组为(42.70±8.62)。实验组与对照组比较,P0.05,差别均具有统计学意义。 3.HE观察结果：对照组3d时角膜部分上皮缺失或空泡状坏死,7d时角膜缘及周边部炎性细胞浸润严重,可见单层内皮细胞围绕而成的毛细血管腔。14d时基质层纤维水肿明显,排列紊乱,可见大量淋巴细胞及新生血管,管腔粗大,排列紊乱。治疗组7d时角膜上皮浅层细胞坏死或变性,胶原纤维肿胀,可见少量空泡,但排列较规整,基质内炎症细胞浸润,角膜新生血管管腔相对较细。14d时角膜上皮完整,角膜基质层结构渐趋整齐,新生血管和炎性细胞稀疏。 4.免疫组织化学检测结果：对照组VEGF阳性表达平均灰度值(862.00±118.67)较实验组VEGF阳性表达平均灰度值(542.00±171.32)增强。差别具有统计学意义(P0.05)。结论：贝伐单抗可有效抑制大鼠碱烧伤早期角膜新生血管的增殖。贝伐单抗滴眼有望成为临床上特异性治疗角膜新生血管安全有效的方法,并有抑制已经形成的新生血管的潜在可能。
[Abstract]:Aim: to investigate the inhibitory effect of bevacizumab on early neovascularization after alkali burn of cornea in rats. Methods: 28 female Wistar rats were randomly divided into experimental group (n = 14) and control group (n = 14). The control group was treated with 0.25% chloramphenicol eye drops and the experimental group with 25mg/ml bevacizumab 0.25% chloramphenicol eye drops twice a day for 14 days. The corneal neovascularization was observed by slit lamp microscope. The number of neovascularization, HE staining and the expression of VEGF protein in corneal tissue were detected by immunohistochemistry. The inhibitory effect on corneal neovascularization was observed. Results: 1. Results under slit lamp microscope: brush neovascularization was observed at the keratoscleral margin on the 3rd day after alkali burn in the cornea of the control group, and neovascularization at the keroscleral edge invaded the cornea on the 5th day. On the 7th day, corneal neovascularization was intertwined into reticular form, and some of the vessels had invaded alkali burn spot, and on the 14th day, corneal neovascularization had become weeping willow and almost occupied the whole cornea. On the 5th day after burn, neovascularization buds were seen in partial quadrant corneal limbus in the treatment group; on the 7th day, the neovascularization invaded the cornea but distributed sparsely and finely; on the 14th day, the neovascularization sparsely distributed outside the alkali burn spot. 2. The number of neovascularization was (137.74 卤21.53) in the control group and (42.70 卤8.62) in the experimental group. The difference between the experimental group and the control group was statistically significant (P 0.05). The results of 3.HE observation showed that the corneal epithelium was absent or vacuolar necrosis in the control group at 3 days, the inflammatory cell infiltration in the limbus and the surrounding part of the cornea was serious at 7 days, and the capillary cavity formed by the monolayer endothelial cells was visible, and the fibroedema in the stromal layer was obvious at 14 days. Disordered, visible a large number of lymphocytes and neovascularization, large lumen, disordered arrangement. In the treatment group, superficial corneal epithelial cells necrosis or degeneration, collagen fiber swelling, a small number of vacuoles, regular arrangement, infiltration of inflammatory cells in the stroma and relatively fine corneal neovascularization lumen were observed in the treatment group at day 14, and the corneal epithelium was intact at 14 days. The structure of corneal stromal layer gradually neatly, and the neovascularization and inflammatory cells were sparse. 4. Immunohistochemical results: the average gray value of VEGF positive expression in the control group (862.00 卤118.67) was higher than that in the experimental group (542.00 卤171.32). The difference was statistically significant (P0.05). Conclusion: bevacizumab can effectively inhibit the proliferation of corneal neovascularization in rats with alkali burn. Bevacizumab eye drops are expected to be a safe and effective method for the specific treatment of corneal neovascularization and have the potential to inhibit the formation of neovascularization.
【学位授予单位】：大连医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R779.1

【参考文献】