Department of Ophthalmology and Visual Sciences, University of Maryland School of Medicine, 10 S. Pine Street, Baltimore, MD 21201, USA.
Invest Ophthalmol Vis Sci. 2011 Jul 29;52(8):5632-40. doi: 10.1167/iovs.10-5983.
To demonstrate corneal endothelial (CE) integrity enhanced during eye banking by a brief treatment of human donor corneoscleral explant (explant) with CE autocrine trophic factor vasoactive intestinal peptide (VIP).
Paired explants were used as control versus VIP (10 nM)-treated before storage in corneal storage medium (4°C). CE ciliary neurotrophic factor receptor (CNTFRα) and CNTF (0.83 nM) responsiveness in connexin 43 upregulation were monitored (Western blot analysis). CE damage in CNTF-modulated explants and corneal buttons from explants was quantified by analysis of panoramic and microscopic images of the alizarin red-stained corneal endothelium. CE cells scraped from the Descemet's membrane were counted. CE VIP receptor was demonstrated (Western blot analysis).
CE cells in every VIP-treated, freshly dissected explant demonstrated higher CNTFRα levels than controls (100% vs. 142% ± 15%; P = 0.014; 7 pairs stored for 4 to 25 days). Nine days after VIP treatment of previously preserved explants, CNTF responsiveness was 174% ± 23% (P = 0.023; 4 pairs) of controls. Panoramic images of explants and corneal buttons revealed that VIP treatment reduced CE damage to 75% ± 6% (P = 0.023; 4 pairs) and 71% ± 11% (P = 0.016; 9 pairs) of controls, respectively, whereas CE damage to 39% (2 pairs) and 23% ± 4% (P < 0.001; 7 pairs), respectively, was revealed in microscopic images. Twenty-one days after VIP treatment of previously preserved explants, CE cell retention was 206% ± 38% (P = 0.008; 14 pairs) of the control. CE cells from human donor corneas expressed VIP receptor VPAC1 (not VPAC2).
CE integrity during eye banking was enhanced by a brief treatment of the explant with the CE autocrine VIP.
通过短暂处理供体角膜-巩膜组织块（组织块）中的角膜内皮（CE）细胞，用 CE 自分泌营养因子血管活性肠肽（VIP）来证明眼库保存过程中 CE 的完整性得到增强。
使用配对组织块作为对照，与 VIP（10 nM）处理后在角膜保存液（4°C）中储存。通过监测细胞连接蛋白 43 上调的睫状神经营养因子受体（CNTFRα）和睫状神经营养因子（CNTF）（0.83 nM）反应性来监测（Western blot 分析）。通过分析茜素红染色的角膜内皮的全景和显微镜图像来量化 CNTF 调节的组织块和组织块中的 CE 损伤。从 Descemet 膜上刮下的 CE 细胞进行计数。通过 Western blot 分析证明 CE VIP 受体。
在每个 VIP 处理的新鲜组织块中，CE 细胞的 CNTFRα水平均高于对照组（100%比 142%±15%；P=0.014；7 对组织块保存 4 至 25 天）。在先前保存的组织块中 VIP 处理 9 天后，CNTF 反应性为对照组的 174%±23%（P=0.023；4 对）。组织块和角膜按钮的全景图像显示，VIP 处理将 CE 损伤降低至对照组的 75%±6%（P=0.023；4 对）和 71%±11%（P=0.016；9 对），而显微镜图像显示 CE 损伤分别为 39%（2 对）和 23%±4%（P<0.001；7 对）。在先前保存的组织块中 VIP 处理 21 天后，CE 细胞保留率为对照组的 206%±38%（P=0.008；14 对）。从供体人眼角膜中分离出的 CE 细胞表达 VIP 受体 VPAC1（而不是 VPAC2）。
通过短暂处理组织块中的 CE 细胞，用 CE 自分泌 VIP 增强眼库保存过程中 CE 的完整性。
