Topical hypochlorite ameliorates NF-κB–mediated skin diseases in mice
Thomas H. Leung, … , Susan J. Knox, Seung K. Kim
Thomas H. Leung, … , Susan J. Knox, Seung K. Kim
Published November 15, 2013
Citation Information: J Clin Invest. 2013;123(12):5361-5370. https://doi.org/10.1172/JCI70895.
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Research Article Genetics

Topical hypochlorite ameliorates NF-κB–mediated skin diseases in mice

Abstract

Nuclear factor-κB (NF-κB) regulates cellular responses to inflammation and aging, and alterations in NF-κB signaling underlie the pathogenesis of multiple human diseases. Effective clinical therapeutics targeting this pathway remain unavailable. In primary human keratinocytes, we found that hypochlorite (HOCl) reversibly inhibited the expression of CCL2 and SOD2, two NF-κB–dependent genes. In cultured cells, HOCl inhibited the activity of inhibitor of NF-κB kinase (IKK), a key regulator of NF-κB activation, by oxidizing cysteine residues Cys114 and Cys115. In NF-κB reporter mice, topical HOCl reduced LPS-induced NF-κB signaling in skin. We further evaluated topical HOCl use in two mouse models of NF-κB–driven epidermal disease. For mice with acute radiation dermatitis, topical HOCl inhibited the expression of NF-κB–dependent genes, decreased disease severity, and prevented skin ulceration. In aged mice, topical HOCl attenuated age-dependent production of p16INK4a and expression of the DNA repair gene Rad50. Additionally, skin of aged HOCl-treated mice acquired enhanced epidermal thickness and proliferation, comparable to skin in juvenile animals. These data suggest that topical HOCl reduces NF-κB–mediated epidermal pathology in radiation dermatitis and skin aging through IKK modulation and motivate the exploration of HOCl use for clinical aims.

Authors

Thomas H. Leung, Lillian F. Zhang, Jing Wang, Shoucheng Ning, Susan J. Knox, Seung K. Kim

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Figure 2

HOCl blocks intracellular NF-κB signaling independent of IκBα.

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HOCl blocks intracellular NF-κB signaling independent of IκBα. (A) Relat...
(A) Relative mRNA levels of Cxcl10 in TNF-α–stimulated mouse wild-type cells with (dotted line) and without (solid line) HOCl pretreatment. (B) Relative fold changes of luciferase activity in LPS-stimulated transgenic wild-type cells carrying an integrated NF-κB reporter gene driving luciferase with (dotted line) and without (solid line) HOCl pretreatment. (C) Relative mRNA transcript levels of Ccl2 mRNA in TNF-α–stimulated p65-deficient cells with (dotted line) and without (solid line) HOCl pretreatment. (D) Visualization of TNF-α–stimulated p65-dsRed nuclear translocation (TNF-α panels; red arrows). HOCl pretreatment blocks TNF-α–stimulated p65-dsRed fusion protein nuclear translocation (HOCl and HOCl + TNF-α panels). (E) Quantification of p65-dsRed nuclear translocation per 100 cells. (F) Relative mRNA transcript levels of Hmox1 in TNF-α–stimulated mouse wild-type cells with (dotted line) and without (solid line) HOCl pretreatment. (G) Western blot analysis of phospho-p38 (P-p38) and total p38 protein in HOCl-treated cells. Black lines indicate a grouping of images from different parts of the same gel. (H) Relative mRNA levels of Cxcl10 in TNF-α–stimulated IκBα-deficient cells with (dotted line) and without (solid line) HOCl pretreatment. (I) Western blot analysis of IκBα and IκBβ protein levels in TNF-α–stimulated wild-type cells with and without HOCl pretreatment. (J) Relative Ccl2 and Hmox1 mRNA transcript levels in TNF-α–stimulated wild-type cells with (gray bars) and without (black bars) H2O2 pretreatment. Data are presented as the average ± SEM.