Kuerten David, Johnen Sandra, Harmening Nina, Souteyrand George, Walter Peter, Thumann Gabriele
Department of Ophthalmology, RWTH Aachen University, Pauwelsstraße 30, 52074, Aachen, Germany.
Graefes Arch Clin Exp Ophthalmol. 2015 Jul;253(7):1061-9. doi: 10.1007/s00417-015-2954-x. Epub 2015 Feb 18.
The purpose of this study was to investigate the effect of recombinant pigment epithelium-derived factor (rPEDF), secreted by ARPE-19 cells transfected with the human PEDF gene and transplanted subconjunctivally in normal and in rabbits in which corneal neovascularization was elicited by a chemical burn.
Twenty grey Chinchilla Bastard rabbits were randomly assigned to four groups; neovascularization was induced in groups A, B, and C by alkali cauterization. Seven days later, group A received no cell implantation, non-transfected ARPE-19 cells were implanted subconjunctivally in group B, and PEDF-transfected ARPE-19 cells were implanted subconjunctivally in groups C and D (non-cauterized). In-vivo rPEDF secretion was analyzed by immunoblotting, and ELISA of extracts of conjunctival tissue samples taken at different time points. Digital photographs acquired on days 7, 14, and 21 after cauterization were evaluated for lead vessel length, vascular invasion area, and overall neovascularization rate.
At days 14 and 21 after cauterization, significant differences were observed between groups A, B, and C in lead vessel length (day 21: 5.91 ± 0.45, 5.11 ± 1.22, 3.79 ± 0.59 mm, repectively), vascular invasion area (day 21: 35.5 ± 8.65, 34.86 ± 4.92, 19.2 ± 5.03 mm(2) respectively), and rate of corneal neovascularization. Compared to controls, neovascularization was reduced by 37.5 % on day 14 and 47 % on day 21. Analysis of conjunctival tissue extracts showed that rPEDF was secreted by the transplanted PEDF-transfected cells.
Subconjunctivally transplanted, PEDF-transfected ARPE-19 cells secrete rPEDF, which inhibits the corneal neovascularization elicited by alkali cauterization.
本研究旨在探讨转染人色素上皮衍生因子（PEDF）基因的ARPE - 19细胞分泌的重组色素上皮衍生因子（rPEDF），结膜下移植到正常兔以及化学烧伤诱导角膜新生血管形成的兔体内后的作用。
将20只灰色青紫蓝杂种兔随机分为四组；A、B、C组通过碱烧伤诱导新生血管形成。7天后，A组不进行细胞植入，B组结膜下植入未转染的ARPE - 19细胞，C组和D组（未烧伤组）结膜下植入转染PEDF的ARPE - 19细胞。通过免疫印迹法分析体内rPEDF分泌情况，并对不同时间点采集的结膜组织样本提取物进行酶联免疫吸附测定（ELISA）。对烧伤后第7、14和21天拍摄的数码照片评估引导血管长度、血管侵入面积和总体新生血管形成率。
在烧伤后第14天和21天，A、B、C组在引导血管长度（第21天：分别为5.91±0.45、5.11±1.22、3.79±0.59 mm）、血管侵入面积（第21天：分别为35.5±8.65、34.86±4.92、19.2±5.03 mm²）和角膜新生血管形成率方面存在显著差异。与对照组相比，第14天新生血管形成减少37.5%，第21天减少47%。结膜组织提取物分析表明，转染PEDF的移植细胞分泌rPEDF。
结膜下移植的转染PEDF的ARPE - 19细胞分泌rPEDF，可抑制碱烧伤诱导的角膜新生血管形成。
