Mansbridge J N, Knapp A M
Psoriasis Research Institute, Palo Alto, California.
J Histochem Cytochem. 1993 Jun;41(6):909-14. doi: 10.1177/41.6.8315281.
We investigated the penetration of Lucifer Yellow into human and murine epidermis in 4-mm punch biopsies by incubation in dye solution. Lucifer Yellow was taken up freely by the dermis but penetrated only slightly into keratinocytes of the basal and suprabasal layers. However, progressive lateral diffusion was observed in the lowest layers of the stratum corneum, extending a distance of 1 mm in 6 hr. Under high magnification, Lucifer Yellow appeared to lie within rather than between corneocytes of this layer. Control samples stained with Lucifer Yellow after sectioning showed no preferential binding of the dye in this region. We concluded that the localization of staining was the result of diffusion from the cut edge of the stratum corneum. Lucifer Yellow penetration was insensitive to PMSF, 1:10 phenanthroline, or N-ethyl maleimide and was also observed in an in vivo injury, indicating that it was not an artifact of proteolytic or degenerative changes. In contrast, horseradish peroxidase failed to penetrate, suggesting molecular size limitation to channel entry. Diffusion of Lucifer Yellow beneath the stratum corneum marks a pathway for the lateral movement of small molecules of potential importance in the normal physiology of the skin, drug delivery, and pathology.
我们通过将4毫米打孔活检组织在染料溶液中孵育,研究了荧光素黄在人及小鼠表皮中的渗透情况。荧光素黄可自由被真皮摄取,但仅少量渗透到基底层和基底上层的角质形成细胞中。然而,在角质层的最底层观察到了渐进性的侧向扩散,6小时内扩散距离达1毫米。在高倍显微镜下,荧光素黄似乎位于该层角质形成细胞内而非细胞之间。切片后用荧光素黄染色的对照样本在该区域未显示出染料的优先结合。我们得出结论,染色的定位是角质层切缘扩散的结果。荧光素黄的渗透对苯甲基磺酰氟、1:10邻菲啰啉或N - 乙基马来酰亚胺不敏感,在体内损伤中也观察到了这种情况,表明这不是蛋白水解或退行性变化造成的假象。相比之下,辣根过氧化物酶未能渗透,提示通道进入存在分子大小限制。荧光素黄在角质层下方的扩散标志着一条小分子侧向移动的途径,这在皮肤的正常生理、药物递送及病理学中可能具有重要意义。