Nintedanib对碱烧伤大鼠角膜新生血管的抑制作用

Inhibitory effect of Nintedanib on corneal neovascularization in rats after corneal alkali burn

目的:探讨尼达尼布(Nintedanib)对大鼠角膜碱烧伤后新生血管的抑制作用。方法:将SD大鼠角膜碱烧伤动物模型随机分成5组,每组各6只。A组、B组与C组分别给予0.2%,0.05%,0.02%的Nintedanib滴眼液滴眼,D组给予0.1%地塞米松滴眼液,E组给予生理盐水滴眼,每组每天4次滴眼治疗,共14 d。分别于碱烧伤后第3,7,14天观察角膜新生血管(corneal neovascularization,CNV)情况,计算CNV面积在角膜面积中的占比(C/N)。于碱烧伤后第14天处死全部大鼠,HE染色大鼠角膜进行组织学观察,并通过免疫荧光法检测各组VEGFR-2和CD31蛋白的表达情况。结果:大鼠角膜碱烧伤后的第3,7,14天,A组、B组、C组和D组的C/N均小于E组,差异均有统计学意义(均P<0.05);B组与D组各时间点C/N面积的差异均无统计学意义(均P>0.05),但这两组与A组比较,或者与C组比较,差异均有统计学意义(均P<0.05)。HE染色及免疫组织化学染色结果显示:角膜碱烧伤后第14天,各组角膜VEGFR-2和CD31蛋白表达增强,与A组、B组、D组比较,C组、E组CNV旺盛致密,炎症细胞更多,角膜水肿跟明显,VEGFR-2和CD31蛋白表达较强,在表达较弱的三组里,A组的CNV最少。结论:Nintedanib对碱烧伤后大鼠CNV形成具有抑制作用,且0.2%Nintedanib的治疗效果最佳。

Objective:To investigate the inhibitory effect of Nintedanib on neovascularization in rats after corneal alkali burn.Methods:Sprague-Dawley(SD)rats were randomly divided into 5 groups,6 rats in each group.The animals in group A,group B and group C were treated with 0.2%,0.05%and 0.02%Nintedanib eye drops,0.1%dexamethasone eye drops in group D,and normal saline drops in group E,4 times daily for 14 days in each group.The corneal neovascularization(CNV)was observed on the 3rd,7th and 14th day after alkali burn,and the ratio of CNV area to corneal area(C/N)was calculated.On the 14th day after alkali burn,all rats were sacrificed.The cornea of rats was subject to HE staining for histological observation and the expression of VEGFR-2 and CD31 proteins was detected by immunofluorescence.Results:On the 3rd,7th and 14th day after corneal alkali burn in rats,the C/N in group A,group B,group C and group D was significantly lower than that in group E(all P<0.05).The CNV area did not significantly differ between group B and group D at each time point(all P>0.05),whereas the CNV area significantly differed between group A and group C(P<0.05).HE staining and immunohistochemistry showed that the expression of VEGFR-2 and CD31 proteins in the rat cornea was significantly un-regulated on the 14th day after alkali burn.Compared with groups A,B and D,the density of CNV in group C and group E was higher,the quantity of inflammatory cells was larger,corneal edema was more evident and the expression of VEGFR-2 and CD31 proteins was up-regulated.The CNV density in group A was least.Conclusion:Nintedanib exerts an inhibitory effect on CNV formation in rats after alkali burn,and 0.2%Nintedanib yields the optimal therapeutic effect.