Retinoic acid improves baseline barrier function and attenuates TNF-α-induced barrier leak in human bronchial epithelial cell culture model, 16HBE 14o

• Published in: PloS one 2020-12, Vol.15 (12), p.e0242536-e0242536
• Main Authors: Callaghan, Patrick J, Rybakovsky, Elizabeth, Ferrick, Bryan, Thomas, Sunil, Mullin, James M
• Format: Article
• Language: English
• Subjects: Anti-Inflammatory Agents - pharmacology; Antiviral agents; Antiviral drugs; Attenuation; Biology and Life Sciences; Bronchi - cytology; Bronchi - drug effects; Bronchi - metabolism; Cell culture; Cell Line; Chronic obstructive pulmonary disease; COVID-19; Cytokine storm; Cytokines; Electrical resistivity; Epithelial cells; Experiments; Extracellular signal-regulated kinase; Humans; Immune system; Inflammation; Inflammation - drug therapy; Inflammation - metabolism; Interleukin; Interleukin 6; Inulin; Lung diseases; Lungs; Mannitol; Medical research; Medicine and Health Sciences; Morbidity; Permeability; Permeability - drug effects; Phosphorylation; Physical Sciences; Physiology; Proteins; Research and Analysis Methods; Respiratory Mucosa - cytology; Respiratory Mucosa - drug effects; Respiratory Mucosa - metabolism; Retinoic acid; Tight Junctions - drug effects; Tight Junctions - metabolism; Tretinoin - pharmacology; Tumor Necrosis Factor-alpha - metabolism; Tumor necrosis factor-α
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0242536

Retinoic acid (RA) has been shown to improve epithelial and endothelial barrier function and development and even suppress damage inflicted by inflammation on these barriers through regulating immune cell activity. This paper thus sought to determine whether RA could improve baseline barrier function and attenuate TNF-α-induced barrier leak in the human bronchial epithelial cell culture model, 16HBE14o- (16HBE). We show for the first time that RA increases baseline barrier function of these cell layers indicated by an 89% increase in transepithelial electrical resistance (TER) and 22% decrease in 14C-mannitol flux. A simultaneous, RA-induced 70% increase in claudin-4 attests to RA affecting the tight junctional (TJ) complex itself. RA was also effective in alleviating TNF-α-induced 16HBE barrier leak, attenuating 60% of the TNF-α-induced leak to 14C-mannitol and 80% of the leak to 14C-inulin. Interleukin-6-induced barrier leak was also reduced by RA. Treatment of 16HBE cell layers with TNF-α resulted in dramatic decrease in immunostaining for occludin and claudin-4, as well as a downward "band-shift" in occludin Western immunoblots. The presence of RA partially reversed TNF-α's effects on these select TJ proteins. Lastly, RA completely abrogated the TNF-α-induced increase in ERK-1,2 phosphorylation without significantly decreasing the TNF-driven increase in total ERK-1,2. This study suggests RA could be effective as a prophylactic agent in minimizing airway barrier leak and as a therapeutic in preventing leak triggered by inflammatory cascades. Given the growing literature suggesting a "cytokine storm" may be related to COVID-19 morbidity, RA may be a useful adjuvant for use with anti-viral therapies.