Corneal endothelial autocrine VIP enhances its integrity in stored human donor corneoscleral explant.
Corneal endothelial autocrine VIP enhances its integrity in stored human donor corneoscleral explant.
复制标题
角膜内皮自分泌 VIP 增强了储存的人类供体角巩膜外植体的完整性。
DOI：
10.1167/iovs.10-5983
复制
发表时间：
2011
影响因子：
4.4
通讯作者：
Molloy,Joseph
中科院分区：
文献类型：
--
作者：
Koh,Shay-WheyM;Gloria,Dante;Molloy,Joseph
目的：为了证明通过用CE自分泌营养因子血管活性肠肽（VIP）对人供体角膜巩膜外植体（外植体）进行短暂处理在眼库中增强的角膜内皮（CE）完整性。将成对的外植体用作对照，与VIP（10 nM）处理的对照相比，在角膜储存培养基（4 ℃）中储存之前。监测CE睫状神经营养因子受体（CNTFRα）和CNTF（0.83 nM）在连接蛋白43上调中的反应性（Western印迹分析）。通过分析茜素红染色的角膜内皮的全景和显微图像，量化CNTF调制的外植体和来自外植体的角膜按钮中的CE损伤。计数从德斯密氏膜刮下的CE细胞。Western blot分析显示CE VIP受体。每个VIP处理的新鲜解剖外植体中的CE细胞均表现出比对照更高的CNTFRα水平（100% vs. 142%±15%; P= 0.014; 7对储存4 - 25天）。VIP处理之前保存的外植体后9天，CNTF反应性为对照组的174%±23%（P= 0.023; 4对）。外植体和角膜按钮的全景图像显示VIP处理分别将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），而显微图像显示CE损伤分别为39%（2对）和23%±4%（P< 0.001; 7对）。VIP处理之前保存的外植体后21天，CE细胞保留率为对照的206%±38%（P= 0.008; 14对）。来自人供体角膜的CE细胞表达VIP受体VPAC 1（而非VPAC 2）。通过用CE自分泌VIP对外植体进行简短处理，增强了眼库保存期间的CE完整性。
Purpose.: 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).Methods.: 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).Results.: 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).Conclusions.: CE integrity during eye banking was enhanced by a brief treatment of the explant with the CE autocrine VIP.