贝伐单抗抑制缝线诱导大鼠角膜新生血管及淋巴管

Inhibition effect of bevacizumab on suture-induced corneal neovascularization and lymph angiogenesis of rats

目的 观察角膜缝线诱导的大鼠角膜新生血管和新生淋巴管生成情况及贝伐单抗(bevacizumab)结膜下注射对其生成的抑制作用。方法 将55只大鼠分为三组:对照组(3只大鼠6只眼,不缝线),生理盐水组(26只大鼠26只眼,角膜缝线后结膜下注射生理盐水0.05 ml)及贝伐单抗组(26只大鼠26只眼,角膜缝线后结膜下注射贝伐单抗0.05 ml)。分别于术后第4,7,10,14天测量术眼角膜新生血管面积,HE染色检测角膜炎症反应情况,免疫组化染色检测VEGF-C及CD31蛋白表达,电镜观察角膜新生血管及淋巴管形成情况。结果 与生理盐水组相比,贝伐单抗组术后角膜新生血管面积明显较小(P<0.05)。大鼠角膜缝线后VEGF-C及CD31蛋白表达的积分光密度值(intergral optical density,IOD),贝伐单抗组较生理盐水组降低(P<0.01);大鼠角膜缝线后VEGF-C及CD31蛋白表达的IOD值在术后14 d时,贝伐单抗组与对照组相比,差异无统计学意义(P>0.05)。透射电镜下可见贝伐单抗组角膜新生血管及淋巴管出现较晚,淋巴管密度较低。结论 贝伐单抗能够减少大鼠角膜缝线后早期新生血管及淋巴管形成,其抑制淋巴管形成的机制可能是阻断VEGF-C/VEGFR-3信号传导通路。

Objective To observe the growth of corneal neovascularization(CNV)and corneal lymphangiogenesis(CL)induced by suture in SD rats and to explore the inhibitory effect of subconjunctival bevacizumab on CNV and CL.Methods Fifty-five SD rats were randomly divided into control group(3 rats 6 eyes,without corneal suture),physiological saline group(PS group,26 rats 26 eyes)and bevacizumab group(26 rats 26 eyes).The eyes were subconjunctivally injected with 0.05 ml physiological saline immediately after corneal suture in PS group.The eyes were subconjunctivally injected with 0.05 ml(1.25 mg)bevacizumab immediately after corneal suture in bevacizumab group.The occurrence and development of CNV and CL were observed and the area of CNV was calculated 4,7,10,14 days after corneal suture.The expression of VEGF-C and CD31 was detected using immunochemistry and the inflammatory reaction in cornea was observed using hematoxylin-eosin staining,and CNV and CL were observed under electron microscope.Results CNV area was statistically smaller in bevacizumab group than in PS group(P<0.05).The intergral optical densities of VEGF-C and CD31 were statistically lower in bevacizumab group than in PS group(P<0.01),but there was no statistically significant difference between control group and bevacizumab group at 14 d after corneal suture(P>0.05).Compared with PS group,the CNV and CL occurred later and the amount of CL was less in bevacizumab group under electron microscope.Conclusion Subconjunctival bevacizumab may inhibit the growth of CNV and CL after corneal suture,and CL may be inhibited by the blocking of VEGF-C/VEGFR-3 signal transduction pathway.