视网膜前体细胞移植对缺血再灌注损伤大鼠视网膜形态影响的实验研究

Effect of retinal precursor cells transplantation on retinal morphology of rats with retinal ischemia-reperfusion injury

目的:研究视网膜前体细胞(RPCs)移植对视网膜缺血再灌注(RIR)损伤大鼠视网膜形态的影响。方法:选取成年健康雄性SD大鼠56只,随机分为七组,每组8只。对照组不做任何处理;假手术组(sham组)只做前房穿刺;视网膜缺血再灌注损伤组(RIR组)为前房灌注模型组;视网膜前体细胞视网膜下腔移植组(RIR+R-C组)前房灌注后在视网膜下腔移植RPCs;磷酸盐缓冲液(PBS)视网膜下腔移植组(RIR+R-PBS组)前房灌注后在视网膜下腔移植PBS;视网膜前体细胞上丘移植组(RIR+SC-C组)前房灌注后在上丘移植RPCs;PBS上丘移植组(RIR+SC-PBS组)前房灌注后在上丘移植PBS。通过免疫荧光染色观察移植细胞在大鼠体内存活、迁徙与分化情况,并比较各组视网膜厚度。结果:移植的RPCs能够在大鼠体内存活至少8周。上丘移植的RPCs未发现迁移到视网膜,视网膜下腔移植的RPCs能够迁移到视网膜外核层并表达视杆细胞标志物视紫红质。RPCs移植后8周,RIR+SC-C组视网膜厚度与RIR+SC-PBS组比较显著增加(P<0.001);RIR+R-C组视网膜厚度与RIR+R-PBS组比较显著增加(P<0.001);RIR+R-C组视网膜厚度与RIR+SC-C组比较显著增加(P<0.001)。结论:视网膜前体细胞移植能够改善视网膜缺血再灌注损伤大鼠视网膜变薄的情况。

Objective:To investigate the effect of retinal precursor cells(RPCs)transplantation on retinal morphology of rats with retinal ischemia-reperfusion(RIR)injury.Methods:Fifty-six adult healthy male SD rats were selected and randomly divided into seven groups,each with 8 rats.The control group did not do any treatment.The sham operation group(sham group)only performed anterior chamber puncture.The RIR injury group(RIR group)was the anterior chamber perfusion model group.In the RPCs subretinal space(SRS)transplantation group(RIR+R-C group),RPCs were transplanted into SRS after anterior chamber perfusion.In the phosphate buffered saline(PBS)SRS transplantation group(RIR+R-PBS group),PBS was transplanted into SRS after anterior chamber perfusion.In the RPCs superior colliculus(SC)transplantation group(RIR+SC-C group),RPCs were transplanted into SC after anterior chamber perfusion.In the PBS SC transplantation group(RIR+SC-PBS group),PBS was transplanted into SC after anterior chamber perfusion.The survival,migration and differentiation of transplanted cells in rats were observed by immunofluorescence staining,and the retinal thickness of each group was compared.Results:The transplanted RPCs could survive in rats for at least 8 weeks.The RPCs transplanted into SC were not found to migrate to the retina.The RPCs transplanted into SRS could migrate to the outer nuclear layer of the retina and express the rod cell marker rhodopsin.8 weeks after RPCs transplantation,the retinal thickness of the RIR+SC-C group increased significantly in comparison with that of the RIR+SC-PBS group(P<0.001);the retinal thickness of the RIR+R-C group increased significantly in comparison with that of the RIR+R-PBS group(P<0.001);the retinal thickness of the RIR+R-C group increased significantly in comparison with that of the RIR+SC-C group(P<0.001).Concusion:Retinal precursor cells transplantation can improve the thinning of the retina in rats with retinal ischemia-reperfusion injury.