ZA-76对大鼠脉络膜新生血管形成、兔眼血流及血管内皮细胞的影响

Effects of ZA-76 on experimental choroidal neovascularization in rats,ocular blood flow in rabbits and human umbilical vein endothelial cell proliferation

目的观察ZA-76对激光诱发大鼠脉络膜新生血管(choroidal neovascularization,CNV)形成、高眼压兔眼血流及人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)增殖的影响。方法氪激光诱发雄性棕色挪威大鼠CNV。30只(60眼)模型大鼠随机分为治疗组和对照组,每组15只(30眼)。治疗组在光凝后第2天开始腹腔注射20g·L-1ZA-76(20mg·kg-1),对照组给予等体积的溶媒二甲基亚砜,每天1次,时间均为4周。光凝后2周、4周分别行眼底照相和眼底荧光血管造影,评估CNV的形成率。光凝后4周处死大鼠,取材作病理切片,用于HE染色及免疫组织化学检测VEGF、COX-2的表达;用彩色微球技术检测20g·L-1ZA-76(50μL)对高眼压兔眼血流的影响;MTT法检测不同浓度ZA-76(100mg·L-1、200mg·L-1、400mg·L-1)对HUVEC细胞增殖的影响。结果治疗组光凝后2周、4周CNV形成率分别为80.83%、79.58%,明显低于对照组的94.58%、95.00%(均为P<0.01);治疗组VEGF、COX-2阳性染色密度分别为(51.13±6.15)mm-2、(37.22±5·18)mm-2,低于对照组[分别为(71.55±9.89)mm-2、(58.78±7.02)mm-2(均为P<0.01)];治疗组脉络膜血流在30min、60min、120min均明显高于对照组;不同浓度ZA-76干预48h,各浓度组ZA-76对HUVEC细胞增殖抑制作用均强于对照组,以400mg·L-1ZA-76作用更显著。结论 ZA-76能抑制激光诱导大鼠CNV形成,抑制VEGF、COX-2的表达;抑制血管内皮细胞增生;改善脉络膜血流。

Objective To investigate the effects of ZA-76 on laser-induced choroidal neovascularization（CNV） in rat,ocular blood flow in high intraocular pressure（IOP） rabbit eyes and human umbilical vein endothelial cell（HUVEC） proliferation.Methods Male brown Norway rats were anesthetized to receive krypton laser to induce CNV.Thirty model rats were divided randomly into control group and treatment group,15 rats（30 eyes） in each group.ZA-76（20 mg·kg-1） with the concentration of 20 g·L-1 and dimethyl sulfoxide were given once-daily through intraperitoneal injection from 1 day after photocoagulation for 4 weeks.The development of CNV was determined by fluorescein angiography at 2 weeks and 4 weeks after photocoagulation.The eyes were enucleated,and the expressions of VEGF and COX-2 were examined by histopathology and immunohistochemistry at 4 weeks after photocoagulation.The colored microsphere technique were used to study the effects of ZA-76（50 μL）on ocular blood flow in high IOP rabbit eyes.The effect of ZA-76 with the concentrations of 100 mg·L-1,200 mg·L-1,400 mg·L-1 on HUVEC proliferation were tested by MTT assay.Results The formed rate of CNV at 2 weeks and 4 weeks after photocoagulation in treatment group were 80.83% and 79.58%,which were obviously lower than that in control group（94.58%,95.00%）,there were significant differences（all P0.01）.Positive staining density of VEGF and COX-2 in treatment group were（51.13±6.15）mm-2 and（37.22±5.18）mm-2,which in control group were（71.55±9.89）mm-2 and（58.78±7.02）mm-2,the treatment group were lower than the control group,there were significant differences（all P0.01）.The choroidal blood flow in treatment group at 30 minutes,60 minutes,and 120 minutes were obviously higher than that in control group.The effects of ZA-76 with different concentrations at 48 hours,especially at 400 mg·L-1,on HUVEC proliferation were stronger than that in control group.Conclusion ZA-76 inhibits the development of CNV induced by laser in rats,decreases the expressions of COX-2 and VEGF,improves the choroidal blood flow in high IOP rabbit eyes and inhibits the HUVEC proliferation.