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Bevacizumab对大鼠角膜新生血管抑制及其超微结构影响的实验研究
Experimental Study on the Inhibition of Corneal Neovascularization with Bevacizumab and Their Ultrastructure
【作者】 林芳；
【导师】 高晓唯；
【作者基本信息】 石河子大学 ， 眼科学， 2009， 硕士
【摘要】 目的:探讨贝伐单抗对角膜新生血管形成及角膜内血管内皮生长因子(VEGF)的影响及其超微结构的影响。方法:盘实验用Wistar大鼠128只随机分4组,采有碱烧伤的方法制备大鼠角膜新生血管模型。正常不烧伤组32只,1、烧伤后隔日球结膜下注射0.1毫升生理盐水组(32只),2、烧伤后隔日球结膜下注射贝伐单抗0.1毫升组(32只),3、烧伤后隔日球结膜下注射地塞米松0.1毫升组(32只)。碱烧伤术后在裂隙灯显微镜下观察大鼠角膜混浊度；宏观测量新生血管长度：组织病理切片HE染色作微血管计数；透射电镜观察超微结构的改变；免疫组化检测角膜VEGF的蛋白表达情况；CD34标记新生血管,显微镜下微血管计数方法研究角膜新生血管形成及抑制情况。结果:治疗组在3、7、10、14天较对照组角膜混浊程度轻(P<0.05)；第14天形成的新生血管数量较对照组少(P<0.05)实验组新生血管微血管数量减少,VEGF蛋白表达下降,具有统计学差异。VEGF主要表达在角膜受损区的感染细胞胞浆内,其出现时间与位置与角膜新生血管一致。贝伐单抗和地塞米松均可有效抑制角膜新生血管,减少角膜内VEGF表达,两者无统计学差异,结膜下注射贝伐单抗和地塞米松后,大鼠角膜超微结构无除烧伤后其它显著改变。结论：VEGF是一种重要的角膜内新生血管形成因子,其变化与角膜新生血管平行;一定剂量贝伐单抗、地塞米松可抑制角膜新生血管,减少角膜内VEGF表达,且两者对角膜的超微结构均无显著药物毒性。。
【Abstract】 AIM:To explore the Bevacizumab therapy for neovascularization of the rat cornea and its impact on the ultrastructureM ethods:established,Wistar rats were selected for four groups at random, Group1 (n=32) normal; Group2 (n=32) received a subconjunctival injection of 0.1ml of 0,9% saline solution; Group3 (n=32) received a subconjunctival injection of 0.1 ml bevacizumab group; Group4 (n=32) received a subconjunctival injection of 0.1 ml dexamethasone group。All groups were observed the corneal opacity by slit-lamp microscope at day,3,7,10,14 after corneal alkali burns.neovascularization were evaluated histological after enucleation,fixation with formaldehyde solution,and staining with hematoxylin and eosin(HE).The contents of blood vessel in corneas were measured.Corneal ultrastructural morphology was observed by transmission electron microscope(TEM).The localization and expression of VEGF,CD34 were determined by immunohistochemical method.Result:lt was found that corneal opacity was slighter in treatment group than that in control group at day 3,7,10,14 after cautery,which there were statistically significant difference between the treatment group and the control group (P<0.05).From the point of neovascularization counted number,the rat corneal neovascularization in the treatment group was less than that in the control group at day 14 after alkali burn (P<0.05).3,7,10,14,with statistically significant difference (P<0.05).The expressions of VEGF,CD34 in normal rat’s cornea were restricted in epithelia and endothelial cell,and VEGF increased gradually at day 3,subsequently the expression decreased slowly after this.There was no difference among bevacizumab and dexamethasone.Conclusion:Bevacizumab can effectively restrain corneal neovascularization and reduce the corneal opacity. VEGF play important parts in alkali burn of the rat cornea. Bevacizumab therapy may ameliorate corneal alkali burn by effecting with those factors. Bevacizumab therapy for alkali burn is safe andutility.