康柏西普和雷珠单抗对大鼠角膜新生血管抑制作用的比较

Comparison of the inhibitory effects of conbercept and ranibizumab on corneal neovascularization in rats

目的研究结膜下注射康柏西普和雷珠单抗对大鼠角膜新生血管(CNV)的抑制作用。方法选用SPF级雄性SD大鼠72只,采用随机数字表法将大鼠分为正常对照组(n=12)、模型对照组(n=20)、雷珠单抗组(n=20)、康柏西普组(n=20)。模型对照组、雷珠单抗组、康柏西普组用碱烧伤法诱导CNV的形成,并于造模后当天分别给予球结膜下注射生理盐水0.05 ml、雷珠单抗0.5 mg(0.05 ml)、康柏西普0.5 mg(0.05 ml)。于造模后4、7、14 d行裂隙灯显微镜拍照,计算CNV面积。每组制作眼球石蜡切片后行苏木精-伊红染色、免疫组化CD34染色检测微血管密度(MVD)。造模后14 d取各组角膜组织,Western blot和实时定量PCR检测角膜组织中血管内皮生长因子(VEGF)、VEGFR-2蛋白及mRNA的表达。结果各组CNV面积结果显示:造模后4、7、14 d雷珠单抗组和康柏西普组CNV面积均小于模型对照组(P<0.05);雷珠单抗组和康柏西普组CNV面积差异无统计学意义(P均>0.05)。苏木精-伊红染色结果显示:造模后4、7、14 d雷珠单抗组和康柏西普组角膜基质水肿程度减轻,CNV数量以及炎性细胞浸润均较模型对照组降低。免疫组化CD34染色检测MVD结果显示:雷珠单抗组和康柏西普组MVD均小于模型对照组(P<0.05);康柏西普组和雷珠单抗组MVD差异无统计学意义(P>0.05)。Western blot和实时定量PCR结果显示:雷珠单抗组和康柏西普组VEGF、VEGFR-2蛋白和mRNA表达均低于模型对照组(P均<0.05)。结论结膜下注射雷珠单抗或康柏西普对大鼠CNV均有抑制作用,且效果相当,两药均可降低角膜组织中VEGF、VEGFR-2的表达。

Objective To compare the inhibitory effect of subconjunctival injection of conbercept and ranibizumab on corneal neovascularization(CNV)in rats.Methods A total of 72 SPF male SD rats were randomly divided into four groups including normal control group(12 rats),model control group(20 rats),ranibizumab group(20 rats)and conbercept group(20 rats).CNV formation was induced by alkali burn in the model control group,ranibizumab group and conbercept group,which received subconjunctival injection of 0.05 ml normal saline,0.5 mg(0.05 ml)ranibizumab,or0.5 mg(0.05 ml)conbercept on the day of modeling,respectively.At 4,7 and 14 days after modeling,the anterior segment photos of the eyes were captured by slit lamp microscope and the area of CNV was calculated.The structure of the corneal tissue was detected by hematoxylin-eosin(H-E)staining and the microvessel density(MVD)was observed by immunohistochemical CD34-positive(CD34^+)staining.On the 14 th day after modeling,Western blot and quantitative real-time PCR(qRT-PCR)were used to observe the expression of vascular endothelial growth factor(VEGF)and vascular endothelial growth factor receptor 2(VEGFR-2)protein and mRNA.Results The CNV area of the ranibizumab group and conbercept group was smaller than that of the model control group(P<0.05)at 4,7 and 14 days after the establishment of the model,and there was no significant difference in CNV area between the ranibizumab group and conbercept group(P>0.05).H-E staining showed that the degree of corneal stromal edema,CNV count and inflam-matory cell infiltration were alleviated in the ranibizumab group and conbercept group compared with the model control group at 4,7 and 14 days after modeling.The results of immunohistochemical CD34^+staining showed that the MVD in the ranibizumab group and conbercept group was less than that in the model control group(P<0.05),but no statistically significant difference was found between the ranibizumab group and conbercept group(P>0.05).Western blot and qRT-PCR results showed that the expression of VEGF and VEGFR-2 protein and mRNA in the ranibizumab group and conbercept group were lower than that of the model control group(P<0.05).Conclusions Subconjunctival injection of ranibizumab or conbercept have equivalent effects on CNV in rats,and ranibizumab or conbercept can reduce the expression of VEGF and VEGFR-2.