Section of Human Anatomy and Histology, Department of Biomedical and Biotechnological Sciences, University of Catania, Catania, Italy.
Eye Clinic, University of Catania, Catania, Italy.
Peptides. 2018 Jan;99:20-26. doi: 10.1016/j.peptides.2017.11.003. Epub 2017 Nov 8.
Cornea's posterior surface includes endothelium maintaining stromal hydration and clarity. Due to their limited proliferative capability, the loss of endothelial cells can outcome in permanent opacity. In the last years, different studies have demonstrated the protective effect of pituitary adenylate cyclase-activating polypeptide (PACAP) in different ocular diseases. However, its role on human corneal endothelial cells (HCECs) has not been investigated, yet. Here, we have developed a culture protocol to differentiate HCECs from donor's cornea. PACAP treatment prevented damage induced by growth factors deprivation of cells grown on transwell supports as revealed by TERR measurements. Moreover, this peptide significantly increased tight junction proteins expression by conferring resistance to endothelial barrier. This effect is also related to promotion of cell viability as demonstrated by MTT assay. Furthermore, PACAP stimulated repairing of corneal endothelium lesion as shown by wound healing analysis. In conclusion, our data suggest that this peptide could represent an important trophic factor in maintaining functionality of human corneal endothelium.
角膜的后表面包括维持基质水合和透明的内皮细胞。由于其有限的增殖能力,内皮细胞的丧失可能导致永久性混浊。在过去的几年中,不同的研究已经证明了垂体腺苷酸环化酶激活肽(PACAP)在不同眼部疾病中的保护作用。然而,其在人角膜内皮细胞(HCEC)中的作用尚未得到研究。在这里,我们开发了一种从供体角膜中分离 HCEC 的培养方案。PACAP 处理可防止在 Transwell 支持物上生长的细胞因生长因子剥夺而受损,这一点通过 TERR 测量得到证实。此外,该肽通过赋予内皮屏障抗性,显著增加了紧密连接蛋白的表达。通过 MTT 测定证实,这种作用也与促进细胞活力有关。此外,PACAP 刺激了角膜内皮细胞损伤的修复,如伤口愈合分析所示。总之,我们的数据表明,这种肽可能是维持人角膜内皮细胞功能的重要营养因子。