标题：Ranibizumab抑制碱烧伤诱导小鼠角膜新生血管作用观察

      作者：张 婷 1，马晓昀 2，张 昀 2，刘 林 3，丁雯芝 1，邹 俊 1，3
    (1.上海交通大学附属第六人民医院眼科，上海 200233；
2.上海市光华中西医结合医院，上海 200052；
3.上海市第十人民医院眼科，上海 200072)

      卷次： 2015年26卷19期

      【摘要】 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-1α mRNA、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-1α mRNA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。

      【关键词】 角膜新生血管；碱烧伤；雷尼单抗；低氧诱导因子-1α；血管内皮生长因子

      【中图分类号】 R-332 【文献标识码】 A 【文章编号】 1003—6350（2015）19—2813—05


Inhibition effect of Ranibizumab on alkali burn induced-corneal neovascularization in mice.
ZHANG Ting 1, MA
Xiao-yun 2, ZHANG Yun 2, LIU Lin 3, DING Wen-zhi 1, ZOU Jun 1,3. 1. Department of Ophthalmology, Shanghai Sixth
People's Hospital, Shanghai Jiaotong University, Shanghai 200233, CHINA; 2. Shanghai Guanghua Hospital of
Integrative Medicine, Shanghai 200052, CHINA; 3. Department of Ophthalmology, Shanghai Tenth People's Hospital,
Shanghai 200072, CHINA

【Abstract】 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-1α mRNA 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-1α mRNA were lower than nega-