目的探索在新西兰兔视网膜中央动脉阻塞(CRAO)模型中行经睫状体平坦部玻璃体切割术(PPV)联合反搏术构建新型视网膜缺血-再灌注(RIR)损伤模型。
方法选用20只健康成年新西兰兔,按照随机数字表法随机分为2个组,每组10只:激光组单纯行视网膜动脉激光光凝术,反搏术组于光凝术后行PPV联合反搏术,均取右眼为实验眼,左眼作为正常对照组。通过荧光素眼底血管造影(FFA)、玻璃体腔氧分压(PO 2)评估灌注恢复情况,观察视网膜电图(ERG)的振荡电位(OPs)变化评估视网膜功能改变,苏木精-伊红染色法观察视网膜结构改变。
结果反搏术组术中即可观察到视网膜灌注恢复;术后2 h,FFA检查示所有眼视网膜动静脉完全恢复灌注,早期即见视网膜动脉充盈,随后静脉充盈,充盈时间无延迟,无血流中断。不同时间点反搏术组、激光组和正常对照组PO 2百分数总体比较,差异均有统计学意义( F 分组=330.87, P<0.001; F 时间=985.70, P<0.001),其中激光后、术后不同时间点反搏术组玻璃体腔PO 2百分数分别为(18.67±6.29)%、(38.82±1.48)%、(57.33±4.25)%、(84.51±3.91)%和(89.20±2.97)%,高于激光组的(23.24±1.95)%、(31.44±3.29)%、(40.21±3.05)%、(43.65±3.82)%和(58.07±2.93)%,差异均有统计学意义(均 P<0.05)。不同时间点反搏术组、激光组和正常对照组OPs百分数总体比较,差异均有统计学意义( F 分组=164.09, P<0.001; F 时间=447.91, P<0.001),其中术后3 d、7 d、2周和1个月反搏术组OPs百分数分别为(47.23±2.73)%、(70.79±3.09)%、(78.39±3.63)%、(76.69±4.08)%和(82.18±1.78)%,较激光组的(46.83±2.89)%、(55.32±1.58)%、(51.08±4.02)%、(52.32±6.59)%和(53.46±6.46)%升高,差异均有统计学意义(均 P<0.05)。反搏术组视网膜内层结构破坏较小,有髓神经纤维层(MFL)结构疏松,大量空泡状改变。激光组MFL、内丛状层、内核层和外丛状层结构紊乱,Müller细胞神经纤维破坏。
结论在新西兰兔CRAO模型中行PPV联合反搏术可构建新型RIR损伤模型。
ObjectiveTo explore the feasibility to establish a novel retinal ischemia reperfusion (RIR) injury model by applying pars plana vitrectomy (PPV) combined with retinal vascular counterpulsation in the central retinal artery occlusion (CRAO) model of New Zealand rabbit.
MethodsTwenty New Zealand adult rabbits were randomly divided into two groups by random number table, 10 for each group.Rabbits in the laser group were treated with retinal laser photocoagulation only, while rabbits in the counterpulsation group were treated with PPV combined counterpulsation.The right eye of each New Zealand rabbit was used as the experimental eye and the left eye was used as the normal control (the normal control group). Fundus fluorescence angiography (FFA) , oxygen partial pressure (PO 2) in vitreous cavity was performed to assess the recovery status of perfusion.Scotopic 3.0 oscillatory potentials (OPs) in electroretinogram (ERG) were used to evaluate the retinal function, and retinal pathological sections were used to evaluate the structural changes in the retina.The use and care of the animals complied with the Statement of the Association for Research in Vision and Ophthalmology (ARVO), and the study was approved by the Animal Research Committee of Zhongshan Ophthalmic Center, Sun Yat-sen University.
ResultsIn the counterpulsation group, retinal reperfusion was observed during counterpulsation processure.FFA examination at 2 hours after counterpulsation showed reperfusion of retinal blood stream in all the eyes.Retinal artery filling, followed by venous filling was observed in the early stage, with no delay in filling and no interruption of blood flow.The percentage of vitreous PO 2 was significantly different among the counterpulsation group, the laser group and the normal control group at different time points ( F group=330.87, P<0.001; F time=985.70, P<0.001). The percentages of vitreous PO 2 in the counterpulsation group at different time points was (18.67±6.29)%, (38.82±1.48)%, (57.33±4.25)%, (84.51±3.91)% and (89.20±2.97)%, which were significantly higher than that in the laser group ([23.24±1.95]%, [31.44±3.29]%, [40.21±3.05]%, [43.65±3.82]% and [58.07±2.93]%) (all at P<0.05). The percentage of OPs was significantly different among the counterpulsation group, the laser group and the normal control group at different time points ( F group=164.09, P<0.001; F time=447.91, P<0.001). The percentages of OPs of counterpulsation group at different time points after laser and after PPV were (47.23±2.73)%, (70.79±3.09)%, (78.39±3.63)%, (76.69±4.08)% and (82.18±1.78)%, which were significanthy higher than that in the laser group ([46.83±2.89]%, [55.32±1.58]%, [51.08±4.02]%, [52.32±6.59]% and [53.46±6.46]%) ( all at P<0.05). There was a lesser damage in inner retinal structure in pathological section in the counterpulsation group.The myelinated nerve fiber layer (MFL) was loose and a mass of vacuolar changes were observed in MFL.The structure of MFL, inner plexiform layer, inner and outer plexiform layer in the laser group were disordered, and the Müller cell nerve fibers were destroyed in the laser group.
ConclusionsA novel type of RIR injury model can be established by applying PPV combined with counterpulsation in the CRAO model of New Zealand rabbit.
温鑫,袁敏而,李成,等. 新型视网膜缺血-再灌注损伤模型的建立及评价[J]. 中华实验眼科杂志,2020,38(07):566-572.
DOI:10.3760/cma.j.cn115989-20190404-00162版权归中华医学会所有。
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