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Role of Nitric Oxide in Wound Healing
Author(s) -
Ishida Hiroshi,
Ray Radharaman,
Ray Prabhati
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.5.a1072-a
Subject(s) - wound healing , nitric oxide , blot , nitric oxide synthase , chemistry , in vitro , western blot , enos , cell migration , andrology , microbiology and biotechnology , biology , biochemistry , medicine , immunology , organic chemistry , gene
Increased nitric oxide (NO) synthesized by NO synthase (NOS) mostly causes cytotoxity; however, cytoprotective effects of NO were also demonstrated. We investigated the role of NO in wound healing in vitro using the cultured monolayer normal human epidermal keratynocytes (NHEK) wound healing model with or without sulfur mustard (SM) treatment. When a wound was created on a confluent NHEK culture by scratching with a pipette tip, cells migration to the wounded area was observed at 4 h after wounding, indicating wound healing. This migration was completely inhibited by 10 ƒÝM SM exposure of cells immediately after wounding. The expression levels of NOSs were studied by Western blotting and fluorescence microscopic analyses using antibodies against nitric oxide synthases, NOS1 (nNOS), NOS2 (iNOS), and NOS3 (eNOS). The level of NOS2 determined by Western blotting increased at 7–16 h after wounding, and SM treatment markedly reduced this effect. The basal (control) level of NOS1 was similar to that of NOS2. However, the NOS1 level continuously decreased after wounding without any recovery up to 24 h. NOS3 was not detectable in either control or wounded cells. Immunofluorescence staining results showed a strong NOS2 expression in the migrating cells at the wounded edge at 7–16 h after wounding. SM treatment reduced the basal level of NOS2 and also suppressed the enhanced expression associated with wounding at 7–16 h. These results suggest that the production of NO by NOS2 may have an important role in human skin wound healing.