促红细胞生成素小干扰RNA对兔角膜新生血管的抑制作用

Inhibition of erythropoiet in siRNA on corneal neovascularization of rabbit

目的:评估促红细胞生成素(erythropoietin,EPO)在兔角膜新生血管发病机制中的作用,并观察促红细胞生成素小干扰RNA(siRNA)对角膜新生血管的抑制作用。方法:健康新西兰白兔22只,随机分为实验组及正常对照组,实验组兔双眼行碱烧伤法建立角膜新生血管模型,造模后每日裂隙灯检查角膜形态学改变并计算新生血管面积,同时自造模当日起右眼每日结膜下注射siRNA 1U组成siRNA治疗组,左眼以control siRNA为实验对照组,分别于术后3、7、14、21d取角膜组织行免疫组化染色,观察EPO的表达情况。结果:实验组最早于碱烧伤后3d可见CNV长入,7~14d生长最为旺盛,免疫组织化学染色见随碱烧伤时间延长,角膜EPO的表达逐渐增加;siRNA治疗后CNV延迟长入,面积较实验对照组明显减小,炎性细胞浸润减轻,且角膜中EPO的表达较前者明显降低,差异有统计学意义(P<0.05)。结论:EPO可能在CNV的发生发展中发挥重要作用,碱烧伤后siRNA的早期干预可能通过影响EPO的表达而抑制CNV的生长。

AIM：To observe the expression of erythropoietin（EPO） on the corneal of rabbit and evaluate the inhibition effect of EPO siRNA on corneal neovascularization（CNV）.METHODS：Totally 22 healthy rabbits were randomly divided into 2 groups,which were experimental group and normal control group.Both eyes of rabbits in experimental group were chosen to establish corneal neovascularization model by alkali burn.The morphologic change of corneal was observed with slit lamp microscope and the area of CNV was calculated every day.After alkali burn,the right eye of the experimental group was accepted EPO siRNA injection under the conjunctiva,and the left eye was assigned to be experimental control group.The corneal with CNV was collected for immunohistochemistry at 3d,7d,14 d,21d after alkali burn,and the expression of EPO was m easured.RESULTS：CNV began growing at the 3d after alkali burn in experimental group,and it was vigorous growing at 7d-14 d period.The result of immunohistochemistry shows that the expression of EPO increased after the operation.Compared with experimental group,the rabbits who were treated by EPO siRNA was found with less neovascularization on their corneal,and the expression of EPO decreased.There were statistical significance between the two group at different time（P0.05）.CONCLUSION：EPO is likely to play an important role on CNV growth,and EPO siRNA can inhibit the growth of CNV by restraining the expression of EPO.