肿瘤抑素Tum5对人脐静脉血管内皮细胞血管生成活性以及碱烧伤诱导大鼠角膜新生血管的抑制作用  

Inhibitory effect of Tum5 on angiogenic activity of human umbilical vein endothelial cells and corneal neovascularization induce by alkali burn in rats

作　　者：郑嘉琳 贾育蓉 郭星艺 张琰 漆晨[1] 孙靖 张红 ZHENG Jia-Lin;JIA Yu-Rong;GUO Xing-Yi;ZHANG Yan;QI Chen;SUN Jing;ZHANG Hong(From the Tianjin Medical University Eye Hospital,Tianjin Medical University Eye Institute,School of Optometry and Ophthalmology of Tianjin Medical University,Tianjin 300384,China)

机构地区：[1]天津医科大学眼科医院天津医科大学眼科研究所天津医科大学眼视光学院,天津市300384

出　　处：《眼科新进展》2018年第5期434-439,共6页Recent Advances in Ophthalmology

基　　金：天津市高等学校科技发展计划项目(编号:20120128);天津市自然科学基金重点项目(编号:14JCZDJC36200)~~

摘　　要：目的研究肿瘤抑素Tum5对人脐静脉血管内皮细胞血管生成活性以及碱烧伤诱导的大鼠角膜新生血管的抑制作用。方法取对数生长期人脐静脉血管内皮细胞分为4组:正常对照组:无任何处理的人脐静脉血管内皮细胞;rAd-GFP组:人脐静脉血管内皮细胞经rAd-GFP感染并培养;rAd-Tum5组:人脐静脉血管内皮细胞经rAd-Tum5感染并培养;rAd-Tum5+VEGF组:人脐静脉血管内皮细胞经rAd-Tum5感染后经外源性VEGF处理。通过CCK-8活性检测试剂盒、Transwell实验、Matrigel实验分别检测细胞增殖率、迁移细胞数及管腔形成情况。取雄性健康清洁级SD大鼠64只采用随机数字表法将大鼠分为:正常对照组、碱烧伤组、碱烧伤+rAd-GFP组、碱烧伤+rAd-Tum5组,每组16只,除正常对照组外其余各组建立角膜碱烧伤模型。其中正常对照组无任何处理,碱烧伤组、碱烧伤+rAd-GFP组、碱烧伤+rAd-Tum5组于烧伤后分别经结膜下注射等量无菌生理盐水、rAd-GFP、rAd-Tum5病毒,记录碱烧伤后第1天、第7天、第14天角膜新生血管相对面积及浸润炎性细胞数。结果 CCK-8实验结果显示rAd-Tum5组较正常对照组、rAd-GFP组细胞增殖率降低(均为P<0.01);而rAd-Tum5+VEGF组细胞增殖率较rAdTum5组显著升高(P=0.004),rAd-Tum5+VEGF组与正常对照组和rAd-GFP组相比差异均无统计学意义(均为P>0.05)。Transwell实验结果显示rAd-Tum5组相对于正常对照组、rAd-GFP组每个视野细胞迁移数显著降低(均为P<0.01);而rAdTum5+VEGF组相对于rAd-Tum5组显著升高(P=0.000),但其迁移能力尚未恢复到正常水平,与正常对照组和rAd-GFP组相比差异均有统计学意义(均为P<0.05)。Matrigel实验结果显示rAd-Tum5组相对于正常对照组、rAd-GFP组每个视野细胞成管数显著降低(均为P<0.01);而rAd-Tum5+VEGF组相对于rAd-Tum5组显著升高(P=0.001)。大鼠角膜碱烧伤后第1-14天,各组角膜新生血管密度及数量逐渐增高,但是碱烧伤+rAd-Tum5Objective To investigate the inhibitory effects of Tum5 on the angiogenesis of human umbilical vein endothelial cells(HUVECs)and alkali-induced corneal neovascularization.Methods HUVECs in logarithmic growth phase were divided into 4 groups,cells with untreated as normal control group,cells with the infection of rAd-GFP virus as rAd-GFP group,cells with the infection of rAd-Tum5 virus as rAd-Tum5 group,and cells with the infection of rAd-Tum5 virus followed by VEGF treatment as rAd-Tum5+VEGF group.Then cell proliferation,migration,and tube formation of HUVECs were examined by CCK-8,Transwell and Matrigel assays,respectively.Sixty-four healthy male SD rats were randomly divided into 4 groups(n=16)by using random number table,and they were normal control group,alkali-burn group,alkali-burn+rAd-GFP group,and alkali-burn+rAd-Tum5 group.The alkali-burn rat model was then established except normal control group,and the normal control group received no treatment,whereas the alkali-burn,alkali-burn+rAd-GFP,and alkali-burn+rAd-Tum5 groups received subconjunctival injection of equal volumes of sterilized saline,rAd-GFP virus,and rAd-Tum5 virus,respectively following the alkaline burn.The relative area of corneal neovascularization and the number of infiltrating inflammatory cells were recorded on day 1,7,and 14 after injection.Results The CCK-8 assay showed that the proliferative rate of rAd-Tum5 group was lower than that of the normal control group and rAd-GFP group(both P<0.01),while rAd-Tum5+VEGF group exhibited a significantly greater cell proliferative capability than rAd-Tum5 group(P=0.004).There were no statistical differences between rAd-Tum5+VEGF group,normal control group and rAd-GFP group(all P>0.05).Transwell assay showed the significantly lower number of migrating cells in the rAd-Tum5 group than those in the normal control group and rAd-GFP group(both P<0.01).The number of migrating cells in rAd-Tum5+VEGF group was higher than those in rAd-Tum5 group(P=0.000);however,the migration capacity had not been restor

关 键 词：血管内皮生长因子 抗血管生成活性 角膜新生血管 肿瘤抑素 转染 

分 类 号：R772[医药卫生—眼科]