三种方法建立外伤性增生性玻璃体视网膜病变的兔模型研究

Establishment of a rabbit model of traumatic proliferative vitreoretinopa⁃thy using three methods

目的:比较三种方法建立外伤性增生性玻璃体视网膜病变(TPVR)兔模型的效果。方法:青紫蓝兔24只,共48只眼,在均造成眼外伤的基础上随机分成4组,每组各12只眼:实验组3组,分别为富血小板血浆(PRP)组[玻璃体腔内注射0.3 mLPRP]、转化生长因子β_(2)(TGF-β_(2))组[玻璃体腔内注射0.3 mLTGF-β_(2)(50μg/L)]和视网膜色素上皮(RPE)组[玻璃体腔内注射0.3 mLRPE细胞(1.5×10^(9)/L)],以及对照组(玻璃体腔内注射0.3 mL生理盐水)。术后每周通过眼底照相和B超观察玻璃体及视网膜的增生情况;术后第4周采用Westernblot法检测玻璃体内视网膜增生过程中α-平滑肌肌动蛋白(α-SMA)的表达,采用冰冻切片法在显微镜下观察各组视网膜组织结构。结果:术后第4周,对照组及3组实验组TPVR的平均分级分别是0、3.67、2.33和2.50,3组实验组病变级别显著高于对照组(P<0.05),PRP组病变级别高于TGF-β_(2)组和RPE组(P<0.05),TGF-β_(2)组与RPE组病变级别差异无统计学显著性(P>0.05);术后第4周,各实验组玻璃体内视网膜α-SMA表达均高于对照组(P<0.05);术后第4周,各实验组视网膜切片显示视网膜增生程度均高于对照组。结论:在造成眼球穿通伤的基础上,玻璃体腔内注射RPE细胞、PRP和TGF-β_(2)均能造成兔TPVR,其中玻璃体内注射PRP的建模效果最明显。

AIM:To compare three different methods for establising a rabbit model of traumatic proliferative vitreoretinopathy(TPVR).METHODS:Twenty-four Chinchilla rabbits,with 48 eyes in total,were randomly divided into 4 groups:platelet-rich plasma(PRP)group(intravitreal injection of 0.3 mL PRP),transforming growth factor-β_(2)(TGF-β_(2))group[intravitreal injection of 0.3 mL TGF-β_(2)(50μg/L)],retinal pigment epithelium(RPE)group[intravitreal injection of 0.3 mL RPE cells(1.5×109/L)],and control group(intravitreal injection of 0.3 mL normal saline).The proliferation of vitreous body and retina was observed weekly after operation by fundus photography and B-mode ultrasound.The expression ofα-smooth muscle actin(α-SMA)during intravitreal retinal proliferation was detected by Western blot at 4 weeks after operation,and the retinal tissue structure of each group was observed under a microscope.RE⁃SULTS:At the 4th week after surgery,the mean grades of TPVR lesions in the control group and the 3 experimental groups were 0,3.67,2.33 and 2.50,respectively.The grades of TPVR lesions in the 3 experimental groups were significantly higher than those in the control group(P<0.05),and the grades of TPVR lesions in PRP group were higher than those in TGF-β_(2) group and RPE group(P<0.05).No significant difference was observed between TGF-β_(2) group and RPE group(P>0.05).At the 4th week after surgery,the expression ofα-SMA in the vitreous retina of the experimental groups was higher than that of the control group(P<0.05).The sections of the experimental group showed a higher degree of retinal proliferation than that of the control group.CONCLUSION:On the basis of causing penetrating eye injuries,intravit real injections of RPE cells,PRP and TGF-β_(2) all caused TPVR in rabbits,and PRP has the most pronounced effect.