【摘要】： 目的：探討貝伐單抗對大鼠角膜堿燒傷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制作大鼠右眼單眼角膜堿燒傷誘導(dǎo)角膜新生血管模型,隨機分為實驗組和對照組,每組各14只。對照組應(yīng)用0.25%氯霉素滴眼液滴眼,實驗組應(yīng)用25mg/ml貝伐單抗+0.25%氯霉素滴眼液滴眼,均為每日兩次,共連續(xù)14天。分別應(yīng)用裂隙燈顯微鏡觀察各組角膜新生血管形成狀況,顯微鏡下記錄新生血管數(shù),HE染色,免疫組織化學(xué)法檢測角膜組織中VEGF蛋白表達(dá)這四種方法,觀察其對角膜新生血管的抑制作用。 結(jié)果：1.裂隙燈顯微鏡下觀察結(jié)果：對照組大鼠角膜堿燒傷后的第3天,角鞏膜緣處可見毛刷狀新生血管的生長；第5天角鞏膜緣處新生血管侵入角膜；第7天角膜新生血管交織成網(wǎng)狀,部分血管已侵入堿燒傷斑；第14天角膜新生血管已呈垂柳狀全部侵入堿燒傷斑幾乎占據(jù)整個角膜。治療組燒傷后第5天,部分象限角膜緣可見新生血管芽；第7天,角膜新生血管侵入角膜但分布稀疏而且細(xì)�。坏�14天,新生血管稀疏分布在堿燒傷斑外側(cè)。 2.顯微鏡下新生血管計數(shù)：對照組新生血管個數(shù)(個／400倍視野)為(137.74±21.53)、實驗組為(42.70±8.62)。實驗組與對照組比較,P0.05,差別均具有統(tǒng)計學(xué)意義。 3.HE觀察結(jié)果：對照組3d時角膜部分上皮缺失或空泡狀壞死,7d時角膜緣及周邊部炎性細(xì)胞浸潤嚴(yán)重,可見單層內(nèi)皮細(xì)胞圍繞而成的毛細(xì)血管腔。14d時基質(zhì)層纖維水腫明顯,排列紊亂,可見大量淋巴細(xì)胞及新生血管,管腔粗大,排列紊亂。治療組7d時角膜上皮淺層細(xì)胞壞死或變性,膠原纖維腫脹,可見少量空泡,但排列較規(guī)整,基質(zhì)內(nèi)炎癥細(xì)胞浸潤,角膜新生血管管腔相對較細(xì)。14d時角膜上皮完整,角膜基質(zhì)層結(jié)構(gòu)漸趨整齊,新生血管和炎性細(xì)胞稀疏。 4.免疫組織化學(xué)檢測結(jié)果：對照組VEGF陽性表達(dá)平均灰度值(862.00±118.67)較實驗組VEGF陽性表達(dá)平均灰度值(542.00±171.32)增強。差別具有統(tǒng)計學(xué)意義(P0.05)。 結(jié)論：貝伐單抗可有效抑制大鼠堿燒傷早期角膜新生血管的增殖。貝伐單抗滴眼有望成為臨床上特異性治療角膜新生血管安全有效的方法,并有抑制已經(jīng)形成的新生血管的潛在可能。
[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 鹵21.53) in the control group and (42.70 鹵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 鹵118.67) was higher than that in the experimental group (542.00 鹵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.
【學(xué)位授予單位】：大連醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R779.1

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