Bigliardi P L, Bigliardi-Qi M, Buechner S, Rufli T
Department of Dermatology, Kantonsspital Basel, Switzerland.
J Invest Dermatol. 1998 Aug;111(2):297-301. doi: 10.1046/j.1523-1747.1998.00259.x.
There is increasing evidence that neurotransmitters play a crucial role in skin physiology and pathology. The expression and production of proopiomelanocortin molecules such as beta-endorphin in human epidermis suggest that an opiate receptor is present in keratinocytes. In this paper we show that human epidermal keratinocytes express a mu-opiate receptor on both the mRNA level and the protein level. Performing polymerase chain reaction with cDNA libraries from human epidermal keratinocytes gave the polymerase chain reaction products of the expected length, which were confirmed as mu-opiate receptors by Southern blot analysis. Using in situ hybridization techniques with a specific probe for mu-opiate receptors we detected the receptor in human epidermis. There was a cytoplasmic expression in all layers of the epidermis, which was more distinct in the suprabasal layers. Immunohistochemistry using the mu-opiate receptor-specific antibody indicates that epidermis expresses protein as well, and that the protein level is more elevated in the basal layer. The correlation between the locations of both mRNA and protein expression in skin indicates that the mu-opiate receptor has not only been transcribed but also has a specific function. To prove a function of the receptor we performed a functional assay using skin organ cultures from human skin transplants. After 48 h incubation with Naloxone or beta-endorphin the expression of the mu-opiate receptor in epidermis was significantly downregulated compared with the control. These results show that a functional receptor indeed exists in human epidermis.
越来越多的证据表明,神经递质在皮肤生理和病理过程中起着至关重要的作用。人表皮中促阿片黑素皮质素分子如β-内啡肽的表达和产生表明角质形成细胞中存在阿片受体。在本文中,我们表明人表皮角质形成细胞在mRNA水平和蛋白质水平上均表达μ-阿片受体。用人表皮角质形成细胞的cDNA文库进行聚合酶链反应,得到了预期长度的聚合酶链反应产物,通过Southern印迹分析证实其为μ-阿片受体。使用针对μ-阿片受体的特异性探针的原位杂交技术,我们在人表皮中检测到了该受体。在表皮的所有层中均有细胞质表达,在基底层以上的层中更为明显。使用μ-阿片受体特异性抗体的免疫组织化学表明表皮也表达蛋白质,并且在基底层中蛋白质水平更高。皮肤中mRNA和蛋白质表达位置之间的相关性表明,μ-阿片受体不仅被转录,而且具有特定功能。为了证明该受体的功能,我们使用来自人皮肤移植的皮肤器官培养物进行了功能测定。与对照相比,用纳洛酮或β-内啡肽孵育48小时后,表皮中μ-阿片受体的表达明显下调。这些结果表明,人表皮中确实存在功能性受体。