CAJ | 학술논문

目的探讨趋化因子受体CCR3拮抗剂在实验性角膜新生血管发生过程中的作用及机制.方法实验研究.以碱烧伤法诱导实验性小鼠角膜新生血管模型;选用动物为54只7～8周龄雄性健康的BABL/c小鼠,随机数字表法分成对照组、实验组及VEGF抗体阳性对照组,每组18只;Real-time PCR法检测CCR3在碱烧伤角膜组织中的动态表达;烧伤后角膜局部应用CCR拮抗剂进行干预,在碱烧伤后2周大体观察及全角膜铺片CD31组织化学染色法检测角膜新生血管发生情况;Real-time PCR和免疫印迹法检测烧伤角膜组织内血管生成相关因子的表达.应用独立样本t检验法比较各组间差异.结果碱烧伤后2周,CCR3拮抗剂干预组角膜新生血管相对值较对照组明显减少.对照组为0.77 ±0.15,实验组为0.51 ±0.03,差异有统计学意义(￡=12.91,P =0.00).免疫印迹检测结果显示角膜组织内VEGF的表达对照组为1.15 ±0.30,实验组为0.91 ±0.24,差异无统计学意义(t=1.08,P =0.34).结论 CCR3信号通路阻滞后能抑制实验性角膜新生血管的发生.
목적 탐토추화인자수체CCR3길항제재실험성각막신생혈관발생과정중적작용급궤제.방법 실험연구.이감소상법유도실험성소서각막신생혈관모형;선용동물위54지7～8주령웅성건강적BABL/c소서,수궤수자표법분성대조조、실험조급VEGF항체양성대조조,매조18지;Real-time PCR법검측CCR3재감소상각막조직중적동태표체;소상후각막국부응용CCR길항제진행간예,재감소상후2주대체관찰급전각막포편CD31조직화학염색법검측각막신생혈관발생정황;Real-time PCR화면역인적법검측소상각막조직내혈관생성상관인자적표체.응용독립양본t검험법비교각조간차이.결과 감소상후2주,CCR3길항제간예조각막신생혈관상대치교대조조명현감소.대조조위0.77 ±0.15,실험조위0.51 ±0.03,차이유통계학의의(￡=12.91,P =0.00).면역인적검측결과현시각막조직내VEGF적표체대조조위1.15 ±0.30,실험조위0.91 ±0.24,차이무통계학의의(t=1.08,P =0.34).결론 CCR3신호통로조체후능억제실험성각막신생혈관적발생.
Objective To explore the effect of CCR3 antagonist on the development of experimental corneal neovascularization.Methods Mouse corneas were burned by NaOH to induce corneal neovascularization.Fifty four clean male BABL/c mice aged 7-8 weeks were divided into control group,CCR3 antagonist group and VEGF antibody positive group according to randomized number table.The gene expression of CCR3 and its ligand eotaxin in burned corneas was examined by Real-time PCR.CCR3 antagonist was locally administrated after alkali injury and the formation of corneal neovascular 2 weeks after injury was examined using a digital camera linked to a slit lamp microscope and corneal whole mount staining with CD31.The mRNA and protein expression of chemokines in burned corneas was detected by Real-time PCR and western blot.Results Compared to control group,CCR3 antagonist treated mice resulted in significantly decreased corneal neovascularization.The related CNV area was 0.51 ± 0.03 in the CCR3 antagonist group,and that in the control group was 0.77 ± 0.15,with significant difference between them (t =12.91,P =0.00).Western blot detection did not show significant difference of VEGF protein expression between two groups.Expression level of VEGF in the CCR3 antagonist group was 0.91 ± 0.24,and that in the control group was 1.15 ± 0.30,showing no significant difference (t =1.08,P =0.34).Conclusions Alkali-induced corneal neovascularization was inhibited by CCR3 antagonist.The mechanism that CCR3 pathway plays an important role in corneal neovascularization needs further exploration.