贝伐单抗联合三氧化二砷对兔角膜碱烧伤诱导角膜新生血管的影响

Effects of avastin combined with As2O3 on rabbits corneal neovascularization induced by alkali burn

目的：探讨贝伐单抗联合三氧化二砷（As2 O3）对兔角膜碱烧伤诱导角膜新生血管（CNV）的影响。方法使用中度碱烧伤法建立48只日本大耳白兔 CNV 动物模型，采用随机数字表法分为四组，A 组于角膜碱烧伤后结膜下注射浓度为1 mg／mL 的 As2 O3注射液0．5 mL ＋0．9％氯化钠注射液0．5 mL，B 组于角膜碱烧伤后结膜下注射浓度为25 mg／mL 的贝伐单抗注射液0．5 mL ＋0．9％氯化钠注射液0．5 mL，C 组于角膜碱烧伤后结膜下注射浓度为1 mg／mL 的 As2 O3注射液0．5 mL ＋25 mg／mL 的贝伐单抗注射液0．5 mL，D 组于角膜碱烧伤后结膜下注射0．9％氯化钠注射液1．0 mL。于碱烧伤后6 d、12 d、15 d、21 d、28 d 时测定各组的CNV 面积，且于烧伤后7 d、14 d、28 d 使用免疫组织化学法测定角膜组织中白细胞介素1β（IL-1β）、血管细胞黏附分子1（VCAM-1）、血管内皮生长因子（VEGF）的表达。结果于碱烧伤后28 d，A 组 CNV 面积、IL-1β、VCAM-1、VEGF 表达率分别为（50．28±30．26）mm2、（6．83±1．74）％、（1．52±0．91）％、（5．02±0．82）％，B组分别为（55．19±20．03）mm2、（6．88±1．94）％、（1．41±0．94）％、（2．91±0．84）％，C 组分别为（10．03±5．98）mm2、（1．96±0．99）％、（0．38±0．57）％、（1．08±0．77）％，D 组分别为（83．18±33．05）mm2、（10．16±2．17）％、（3．74±0．83）％、（7．69±1．33）％，四组差异均有统计学意义（F ＝25．33、3．64、13．13、7．23，均 P ＜0．05）；A、B、C 组的 CNV 面积和 IL-1β、VCAM-1、VEGF 的表达均显著性低于 D 组（qcnv ＝40．294、34．281、89．590，qIL-1β＝4．078、4．017、10．043，qVCAM-1＝2．719、2．854、5．791，qVEGF ＝3．270、8．238、11．392，均 P ＜0．05）；A、B 组的 CNV 面积和 IL-1β、VCAM-1、VEGF 的表达均显著性低于 C 组（qCNV ＝49．296、55．309，qIL-1β＝5．965、6．026，qVCAM-1＝1．965、1．775，qVEGF ＝6．790、22．413，均 P ＜0．05）；A、B 组的 CNV 面积和 IL-1β、VCAM-1的表达差异无统计学意义（P ＞0．05）；A 组 VEGF 的表达显著高于 B 组（q ＝4．03，P ＜0．05）。结论贝伐单抗联合 As2 O3可有效抑制兔角膜碱烧伤诱发 CNV 的生长，降低 CNVIL-1β、VCAM-1、VEGF 的表达，比单独用药有更好的疗效，为 CNV 防治开辟了一个新思路。

Objective To investigate the effects of avastin combined with As2 O3 on rabbits corneal neovas-cularization （CNV）induced by alkali burn.Methods A total of 48 Japanese rabbits were induced the CNV models by middle alkali burns.The rabbits were randomly divided into four groups.A group was subconjunctivally injected 0.5mL 1mg/mL of As2 O3 and 0.5mL normal sodium,B group was subconjunctivally injected 0.5mL 25mg/mL of avastin and 0.5mL normal sodium,C group was subconjunctivally injected 0.5mL 1mg/mL of As2 O3 and 0.5mL 25mg/mL of avastin,D group was subconjunctivally injected 1.0mL normal sodium,after corneal alkali burn,respec-tively.CNV area at 6 days,12 days,15 days,21 days and 28 days after burns were calculated,the expression of IL -1β,VCAM-1 and VEGF in corneal tissue at 7 days,14 days,28 days after burns were detected by immunohisto-chemical staining.Results Alkali burn after 28 d,the CNV,IL -1β,VCAM-1,VEGF expression rates of A group were （50.28 ±30.26）mm2 ,（6.83 ±1.74）%,（1.52 ±0.91）%,（5.02 ±0.82）%,respectively,which of B group were （55.19 ±20.03）mm2 ,（6.88 ±1.94）%,（1.41 ±0.94）%,（2.91 ±0.84）%,respectively,which of C group were （10.03 ±5.98）mm2 ,（1.96 ±0.99）%,（0.38 ±0.57）%,（1.08 ±0.77）%,respectively,which of D group＆nbsp;were （83.18 ±33.05）mm2 ,（10.16 ±2.17）%,（3.74 ±0.83）%,（7.69 ±1.33）%,respectively.The differences among four groups were statistically significant （F =25.33,3.64,13.13,7.23,all P 〈0.05 ）.Compared with D group,the CNV area and the expressions of IL -1β,VCAM-1 and VEGF were significantly decreased in A,B,C group（qCNV =40.294,34.281,89.590,qIL -1β =4.078,4.017,10.043,qVCAM-1 =2.719,2.854,5.791,qVEGF =3.270,8.238,11.392,all P 〈0.05）;Compared with C group,the CNV area and the expressions of IL -1β,VCAM-1 and VEGF were significantly decreased in A,B group （qCNV =49.296,55.309;qIL -1β =5.965,6.026;qVCAM-1 =1.965,1.775;qVEGF =6.790,22.413,all P 〈0.05）;Compared with B group,the CNV area and the expressions of IL -1βand VCAM-1 were not significantly different in A group （P 〉0.05）;Compared with B group,the expression of VEGF was significantly increased in A group （q =4.03,P 〈0.05）.Conclusion Avastin combined with As2 O3 can inhibit the growth of CNV induced by alkali burning in rabbits effectively,and reduce the expressions of IL -1β, VCAM-1 and VEGF in CNV.It has better efficacy than single drug,and it opens up a new way for the prevention and treatment of CNV.