摘要
目的观察肝细胞生长因子(HGF)对视网膜色素上皮(RPE)细胞屏障功能的影响以及RPE内过度表达HGF导致视网膜脱离(RD)的病理机制。方法编码HGF(AdCMV.HGF)、绿色荧光蛋白(AdCMV.GFP)的E1/E3缺失的腺病毒载体,以5×10^4噬斑形成单位(pfu)/眼注射到成年有色兔的视网膜下。检查注射后3、7、14、28d时的眼底及组织病理变化,利用免疫组织化学和酶联免疫吸附试验(EusA)3-法检测HGF在视网膜和玻璃体的表达水平。结果对照组注射AdCMV.GFP眼显示GFP几乎仅表达于PRE单核细胞层,AdCMV.HGF注射眼在注射点处的PRE细胞出现强的HGF免疫阳性反应。玻璃体内HGF的表达水平在注射7d后达到最高峰、28d后降低到基础水平。在HGF的表达期内AdCMV.HGF注射眼出现慢性RD和脉络膜慢性炎症。在RD区域,视网膜下的空间内可见增生性的RPE细胞,部分实验兔眼还产生多层的细胞膜结构。结论RPE内过度表达的HGF能引发慢性浆液性RD,同时伴有视网膜下RPE增生。提示HGF可能作为治疗RD的作用靶点。
To investigate the effect of hepatocyte growth factor (HGF) on the barrier function of retinal peigment epithelium (RPE) and to detect the pathological mechanism of retinal detachment (RD) induced by over-expression of HGF in RPE. Methods Sub-retina injection of E1/E3-deleted adenoviral vectors encoding HGF (Ad CMV. HGF) and green fluorescent protein (Ad CMV. GFP) in adult pigmented rabbits [5× 10^4 plaque-forming units (pfu)/eye] to set up the model of retinal detachment. The ocular fundus and pathological changes were observed 3, 7, 14, and 28 days after injection. The expression level of HGF in retina and vitreous body was detected by immunohistochemistry and enzyme linked immunosorbent assay (ELISA). Results In the control eyes injected with AdCMV. GFP, expression of GFP only detected in RPE monolayer. The eyes injected with AdCMV. HGF had strong HGF immune positive action in RPE cells at the injection site. The expression level of HGF in vitreous body reached the peak 7 days after injection and decreased to the basic level 28 days after injection. Chronic RD and chronic choroidal inflammation were found in the eyes injected with AdCMV. HGF within the time frame of HGF expression. Proliferative RPE cells were found in subretinal space in the region of RD, and multilayered cellular membranes developed in some eyes. Conclusion Over-expression of HGF in RPE may induce chronic serous RD with subretinal proliferation of RPE, which suggests that HGF should be further studied as a target for therapeutic intervention in RD.
出处
《中华眼底病杂志》
CAS
CSCD
北大核心
2007年第3期193-197,共5页
Chinese Journal of Ocular Fundus Diseases