NEUROTROPHIC INFLUENCES ON CORNEAL EPITHELIAL-CELLS
NEUROTROPHIC INFLUENCES ON CORNEAL EPITHELIAL-CELLS
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DOI：
10.1006/exer.1994.1145
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发表时间：
1994-11-01
影响因子：
3.4
通讯作者：
BELMONTE, C
中科院分区：
文献类型：
--
作者：
GARCIAHIRSCHFELD, J;LOPEZBRIONES, LG;BELMONTE, C
神经支配在角膜结构和功能的维持中起着重要的作用。角膜神经中所含的神经肽可能参与了这些作用。本研究旨在探讨三叉神经和交感神经以及神经肽对角膜上皮细胞体外增殖活性的影响。将成年兔角膜上皮细胞单独或与三叉神经节感觉神经元或新生兔颈上神经节交感细胞共培养。神经元和角膜上皮细胞共享培养基，但缺乏物理接触。共培养2天或7天后，或P物质（SP）和/或降钙素基因相关肽（CGRP）处理后，用血细胞计测定角膜细胞数量，闪烁计数分析h -3-胸腺嘧啶与角膜上皮细胞的掺入情况。角膜上皮细胞与三叉神经共培养2 d后，细胞数量和h -3-胸腺嘧啶掺入量分别增加了3倍和2.5倍（P < 0.01; P < 0.05），但共培养7 d后，细胞数量和h -3-胸腺嘧啶掺入量的增加趋势逆转。共培养7 d后，交感神经元对角膜上皮细胞增殖有明显的刺激作用（P < 001）。单次给药SP （10 μ M）在1小时后产生h -3-胸腺嘧啶掺入增加，24小时后角膜上皮细胞数量增加。重复施用低浓度SP （0.1 μ M， 24小时6次）也可获得相同的效果。CGRP （1 μ M）对角膜细胞有丝分裂活性有抑制作用。同时给予0.1 μ M的SP可逆转这一效应。这些结果表明，三叉神经和交感神经细胞在体外调节角膜上皮细胞的增殖和分化。SP对上皮细胞有刺激作用，而CGRP对有丝分裂活性有抑制作用，提示感觉神经末梢所含多肽可能参与神经对上皮细胞增殖的影响。
Innervation plays an important role in the maintenance of corneal structure and function. Neuropeptides contained in corneal nerves may be involved in these effects. This study was aimed to determine the influence of trigeminal and sympathetic neurons and of neuropeptides on proliferative activity of corneal epithelial cells in vitro.Corneal epithelial cells from adult rabbits were cultured alone or cocultured either with sensory neurons from the trigeminal ganglion or sympathetic cells from the superior cervical ganglion of newborn rabbits. Neurons and corneal epithelial cells shared the culture medium but lacked physical contact. After 2 or 7 days of coculture, or after treatment with substance P (SP) and/or calcitonin gene-related peptide (CGRP), the number of corneal cells was determined with a haemocytometer, and the incorporation of H-3-thymidine into corneal epithelial cells analysed by scintillation counting.Cell number and H-3-thymidine incorporation increased 3 and 2.5 times respectively (P < 0.01; P < 0.05), after 2 days of coculture of corneal epithelial cells with trigeminal neurons but this increase was reversed after 7 days of coculture. Sympathetic neurons had a stimulating effect on corneal epithelial cell proliferation that was apparent after 7 days of coculture (P < 001). A single administration of SP (10 mu M) produced an increase in H-3-thymidine incorporation after 1 hr and an enhanced number of corneal epithelial cells 24 hr later. The same effect could be obtained by repeated administration of lower concentrations of SP (0.1 mu M, six times over a 24 hr period). CGRP (1 mu M) inhibited mitotic activity of corneal cells 12 hr after treatment. The effect could be reversed by simultaneous administration of SP (0.1 mu M).These results indicate that trigeminal and sympathetic neurons modulate proliferation and perhaps differentiation of corneal epithelium cells in vitro. A stimulating effect was also obtained with SP while mitotic activity was inhibited by CGRP, thus suggesting that peptides contained in sensory nerve terminals may contribute to neural influences on epithelial cell proliferation.