Ranibizumab抑制碱烧伤诱导小鼠角膜新生血管作用观察

摘要

Objective To observe the inhibition effect of Ranibizumab on alkali burn induced-corneal neo-vascularization (CorNV) in mice. Methods Thirty-six healthy BALB/c mice received 1 mol/L NaOH for 30 s to establish models of alkali induced-CorNV of the right eyes. All the mice were randomly divided into two groups, negative control group (eighteen cases, the right eyes received subconjuctival injections of 0.9%normal saline), Ra-nibizumab treatment group (eighteen cases, the right eyes received subconjuctival injections of 10 mg/ml Ranibi-zumab). On days 2, 8 after alkali burn, CorNV was observed and photographed on days 4, 7, 14. The areas of CorNV were calculated. On days 7, 14, hematoxylin-eosin (HE) and immunohistochemical (IHC) staining were con-ducted to observe histopathological change and to detect corneal hypoxia-inducible factor-1α(HIF-1α) and vascu-lar endothelial growth factor (VEGF) expression. Quantitative real-time polymerase chain reaction (real-time PCR) was applied to detect the expression of HIF-1αmRNA and VEGF-A mRNA. Results Frontal section photograph and HE staining revealed that on days 7 and 14 after alkali burn, areas of CorNV in Ranibizumab treatment group were significantly lower than that in negative control group (P<0.05). Real-time PCR showed that relative expres-sion levels of VEGF-A mRNA in Ranibizumab treatment group were all significantly lower than negative control group on day 7 and day 14 after alkali burn, and relative expression levels of HIF-1αmRNA were lower than nega-tive control group on day 7 (all P<0.05). IHC staining revealed that on days 7 and 14 after alkali burn, VEGF-A pro-tein expression in Ranibizumab treatment group decreased compared to negative control group (P<0.05), while HIF-1αprotein expression did not show any significant change. Conclusion Ranibizumab has some inhibition effect on mouse CorNV induced by alkali burn through down-regulation of VEGF.%目的 观察雷尼单抗(Ranibizumab)抑制碱烧伤诱导小鼠角膜新生血管(CorNV)的作用.方法 取清洁级BALB/c小鼠36只,将加有NaOH溶液(1 mol/L)的直径2 mm圆形滤纸片轻贴角膜中央烧灼30 s,制备角膜碱烧伤CorNV模型:右眼接受碱烧伤处理,左眼不予处理.后随机将小鼠分为两组,于碱烧伤后2 d、8 d时右眼分别行球结膜下注射0.9%生理盐水(阴性对照组,18只小鼠)、10 mg/ml Ranibizumab (Ranibizumab治疗组,18只小鼠).碱烧伤后4 d、7d、14 d时行眼前节照相大体观察CorNV生长变化并计算CorNV面积;分别于碱烧伤后7 d、14 d时行苏木素-伊红(HE)染色观察角膜组织病理结构及CorNV变化;免疫组织化学(IHC)染色检测角膜HIF-1α、VEGF-A蛋白表达;实时荧光定量聚合酶链式反应(Real-time PCR)检测角膜HIF-1αmRNA、VEGF-A mRNA表达.结果 眼前节照相、HE染色结果显示:碱烧伤后7 d、14 d时Ranibizumab治疗组较阴性对照组角膜新生血管减少,细小稀疏,CorNV面积明显减小,差异均有统计学意义(P<0.05).PCR结果显示:Ranibizumab治疗组中角膜VEGF-A mRNA相对表达于7 d、14 d时均较阴性对照组降低,7 d时HIF-1αmRNA相对表达降低;差异均有统计学意义(P<0.05).免疫组化结果显示:Ranibizumab治疗组VEGF-A蛋白表达于7 d、14 d时较阴性对照组均降低,差异均有统计学意义(P<0.05);HIF-1α表达未明显改变.结论 Lucentis通过下调VEGF对碱烧伤诱导小鼠角膜CorNV具有一定抑制作用.