基因转染的虹膜色素上皮细胞移植后RCS鼠视网膜BDNF表达观察

Retinal BDNF expressions in RCS rats after transplantation of gene transfected iris pigment epithelium

目的探讨脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)基因转染的虹膜色素上皮细胞(AAV-BDNF-IPE)移植入皇家外科学院(royal college of surgeons,RCS)大鼠视网膜下腔后,不同时期视网膜组织BDNF表达变化。方法通过外路途径将BDNF基因转染的虹膜色素上皮细胞移植到RCS大鼠视网膜下腔,术后3、5、7、9、11周分别取RCS大鼠手术眼及对照组动物眼视网膜组织,用酶联免疫吸附法(Elisa)检测视网膜组织中BDNF的表达水平,比较分析这些数据。结果对照组RCS大鼠出生后3周龄时视网膜组织中BDNF仍保持较高水平,其后迅速降低,其中3周龄组与其它周龄组比较,P<0.01;手术组RCS大鼠术时、术后3、5、7、9、11周各组间两两比较,BDNF表达无显著差异(P>0.05);出生后6周龄直到14周龄的不同时期,AAV-BDNF-IPE移植手术组RCS大鼠视网膜BDNF表达水平均明显高于对照组(其中6周龄组P<0.05,其它各周龄组P<0.01)。结论 BDNF基因转染的虹膜色素上皮细胞在RCS大鼠视网膜下腔移植后,视网膜组织中BDNF可以持续稳定高水平表达,这为临床开发新的神经营养因子给药方式提供了实验依据。

Objective To investigate the retinal brain derived neurotrophic factor（BDNF） expressions in different phases of royal college of surgeons（RCS） rats after BDNF transfected iris pigment epithelium（AAV-BDNF-IPE） being transplanted into the subretinal space of RCS rat.Methods AAV-BDNF-IPEs were transplanted into the subretinal space of RCS rats.BDNF expressions in retinal tissue of intact RCS rats and surgery RCS rats were detected by enzyme linked immunosorbent assay（Elisa） at 3,5,7,9and 11 weeks after surgery.ResultsBDNF expressions in retinal tissue of intact RCS rats were still high at postnatal 3w and were sharply decreased into low level later;retinal BDNF expression of intact RCS rats at postnatal 3w were much higher than those at other postnatal weeks,P0.01.There was no difference in BDNF expressions of surgery RCS rats during surgery and at 3,5,7,9and 11 weeks after surgery,P0.05.BDNF expression in retinal tissue of surgery RCS rats were significantly higher than those in intact rats at postnatal 6,8,10,12 and 14 weeks,（P0.05 at postnatal 6w and P0.01 at postnatal 8,10,12 and 14 weeks,respectively）.Conclusion AAV-BDNF-IPE transplantation into the subretinal space of RCS rats can continuously improve the expression of BDNF in retinal tissue,which exerts a protective effect on the congenital retinal degeneration of RCS rats.