Laboratório de Biologia Celular, Departamento de Ciências Básicas da Saúde, Universidade Federal de Ciências da Saúde de Porto Alegre (UFCSPA), 90050-170 Porto Alegre, RS, Brasil.
Département de Microbiologie-Infectiologie et d'Immunologie, Faculté de Médecine, Université Laval, Québec city, QC G1V 0A6, Canada.
Biochem Cell Biol. 2023 Jun 1;101(3):259-266. doi: 10.1139/bcb-2022-0058. Epub 2023 Jan 19.
The human epidermal melanocyte (hEM) are melanin-producing cells that provide skin pigmentation and protection against ultraviolet radiation. Although purinergic signaling is involved in skin biology and pathology, the presence of NTPDase members, as well as the rate of nucleotides degradation by melanocytes were not described yet. Therefore, in this study, we analyzed the expression of ectonucleotidases in hEM derived from discarded foreskin of male patients. The expression of purinergic enzymes was confirmed by mRNA and flow cytometry. Among the ectonucleotidases, ectonucleoside triphosphate diphosphohydrolase1 (NTPDase1) and ecto-5´-nucleotidase were the ectoenzymes with higher expressions. The hydrolysis rate for ATP, ADP, and AMP was low in comparison to other primary cells already investigated. The amount of ATP in the culture medium was increased after a scratch wound and decreased to basal levels in 48 h, while the NTPDase1 and P2X7 expressions increased. Therefore, it is possible to suggest that after cell injury, the ATP released by hEM into the extracellular space will be hydrolyzed by ectonucleotidases as the NTPDase1 that will control the levels of nucleotides in the skin micro-environment.
人类表皮黑素细胞（hEM）是产生黑色素的细胞，为皮肤提供色素沉着和抵御紫外线辐射。尽管嘌呤能信号参与皮肤生物学和病理学，但尚未描述 NTPDase 成员的存在以及黑素细胞中核苷酸的降解速度。因此，在这项研究中，我们分析了来自男性患者废弃包皮的 hEM 中核苷酸酶的表达。嘌呤能酶的表达通过 mRNA 和流式细胞术得到了证实。在这些外核苷酸酶中，核苷酸三磷酸二磷酸水解酶 1（NTPDase1）和外 5′-核苷酸酶是表达较高的外酶。与已研究的其他原代细胞相比，ATP、ADP 和 AMP 的水解率较低。划痕伤后培养介质中 ATP 的量增加，并在 48 小时后降至基础水平，而 NTPDase1 和 P2X7 的表达增加。因此，可以推测，在细胞损伤后，hEM 释放到细胞外空间的 ATP 将被 NTPDase1 等外核苷酸酶水解，从而控制皮肤微环境中核苷酸的水平。
