Weak Ultraviolet B Enhances the Mislocalization of Claudin-1 Mediated by Nitric Oxide and Peroxynitrite Production in Human Keratinocyte-Derived HaCaT Cells

• Published in: International journal of molecular sciences 2020-10, Vol.21 (19), p.7138
• Main Authors: Kobayashi, Mao, Shu, Shokoku, Marunaka, Kana, Matsunaga, Toshiyuki, Ikari, Akira
• Format: Article
• Language: English
• Subjects: Apoptosis; Barriers; Cadaverine - analogs & derivatives; Cadaverine - pharmacology; Calcium (intracellular); Calcium ions; Calcium oxide; Capsaicin receptors; Cell Survival - radiation effects; Clathrin; Claudin-1 - metabolism; Disruption; Electrical resistivity; Endocytosis; Endocytosis - drug effects; Fluorescence; HaCaT Cells; Health aspects; Humans; Inhibitors; Internalization; Irradiation; Keratinocytes - metabolism; Localization; Mammals; Nitric oxide; Nitric Oxide - metabolism; Nitric Oxide Synthase - metabolism; Nitric-oxide synthase; Oxidative stress; Peroxynitrite; Peroxynitrous Acid - biosynthesis; Phosphorylation; Phosphorylation - radiation effects; Physiological aspects; Proteins; Signal transduction; Signal Transduction - drug effects; Signal Transduction - radiation effects; siRNA; Skin; T cells; Tight Junctions - metabolism; Tight Junctions - radiation effects; Transient receptor potential proteins; TRPV Cation Channels - metabolism; Tyrosine; Ubiquitination; Ubiquitination - radiation effects; Ultraviolet radiation; Ultraviolet Rays
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms21197138

A tight junction (TJ) makes a physical barrier in the epidermal cells of skin. Ultraviolet (UV) light may disrupt the TJ barrier, but the mechanism has not been well clarified. Weak UVB (5 mJ/cm ) caused mislocalization of claudin-1 (CLDN1), a component of the TJ strand, and disruption of TJ barrier in human keratinocyte-derived HaCaT cells. The UVB-induced mislocalization of CLDN1 was inhibited by monodansylcadaverine (MDC), a clathrin-dependent endocytosis inhibitor, suggesting that UVB enhances the internalization of CLDN1. Transepidermal electrical resistance and paracellular flux of lucifer yellow, a fluorescent hydrophilic marker, were rescued by MDC. UVB changed neither the total nor phosphorylation levels of CLDN1, but it increased both mono-ubiquitination and tyrosine nitration levels of CLDN1. Fluorescence measurements revealed that UVB increased intracellular free Ca , nitric oxide (NO), and peroxynitrite contents, which were inhibited by Opsin2 (OPN2) siRNA, suggesting that OPN2 functions as a UVB sensor. The effects of UVB were inhibited by an antagonist of transient receptor potential type vanilloid 1 (TRPV1) and Ca chelator. Both NO donor and peroxynitrite donor induced the mislocalization of CLDN1 and disruption of TJ barrier, which were rescued by a NO synthase (NOS) inhibitor and a peroxynitrite scavenger. Weak UVB irradiation induced the disruption of TJ barrier mediated by mislocalization of CLDN1 in HaCaT cells. The OPN2/TRPV1/NOS signaling pathway may be a novel target for preventing destruction of the TJ barrier by UVB irradiation.