IL-4 promotes asymmetric dimethylarginine accumulation, oxo-nitrative stress, and hypoxic response–induced mitochondrial loss in airway epithelial cells

• Published in: Journal of allergy and clinical immunology Vol. 138; no. 1; pp. 130 - 141.e9
• Main Authors: Pattnaik, Bijay, MSc, Bodas, Manish, PhD, Bhatraju, Naveen Kumar, PhD, Ahmad, Tanveer, PhD, Pant, Richa, MSc, Guleria, Randeep, MD, DM, Ghosh, Balaram, PhD, Agrawal, Anurag, MD, PhD
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01-07-2016; Elsevier Limited
• Subjects: ADMA; Allergy and Immunology; Amidohydrolases - metabolism; Apoptosis; Arginine - analogs & derivatives; Arginine - metabolism; Asthma; Asthma - etiology; Asthma - metabolism; Asthma - pathology; Binding sites; Calpain - metabolism; Cell Line; Cells, Cultured; Cytokines; Cytokines - metabolism; Diet; Epithelial Cells - drug effects; Epithelial Cells - metabolism; Flow cytometry; Humans; Hypoxia - metabolism; hypoxic response; IL-4; Immunoglobulins; Inflammation; Inflammation Mediators - metabolism; Interleukin-4 - metabolism; Interleukin-4 - pharmacology; Metabolism; mitochondria; Mitochondria - drug effects; Mitochondria - metabolism; mitochondrial transcription factor A; Nitric Oxide; Oxidative Stress; Pathogenesis; Peroxynitrous Acid - metabolism; Proteins; Proteolysis; Reactive Oxygen Species - metabolism; Respiratory Mucosa - metabolism; Rodents; Transcription factors; eNOS; HFA; Reactive oxygen species; HRP; Horseradish peroxidase; HIF-1α; AHR; PRMT; hypoxic response; Dimethylarginine dimethylaminohydrolase; HFR; Inducible nitric oxide synthase; Asymmetric dimethylarginine; iNOS; Mitochondrial reactive oxygen species; mitochondrial transcription factor A; mtROS; Peroxisome proliferator-activated receptor γ coactivator 1α; 4′,6-Diamidino-2-phenylindole; Ovalbumin; Hypoxia-inducible factor 1α; DAB; mitochondria; IL-4; High fructose; ADMA; AAI; DAPI; PGC-1α; Airway hyperresponsiveness; Allergic airway inflammation; Endothelial nitric oxide synthase; High fat; Protein-arginine methyl transferase; ROS; DDAH; TFAM; Diaminobenzidine; OVA
• ISSN: 0091-6749; 1097-6825
• DOI: 10.1016/j.jaci.2015.11.036

Background Obesity is known to increase asthma risk and severity. Increased levels of asymmetric dimethylarginine (ADMA), an endogenous nitric oxide synthase inhibitor, are associated with mitochondrial toxicity, asthma, and metabolic syndrome. IL-4 upregulates the expression of protein arginine methyltransferases, which are essential for ADMA formation. Importantly, cross-talk between IL-4, ADMA, and mitochondrial dysfunction could explain how obesity and IL-4 can synergize to exacerbate allergic inflammation. Objective We sought to investigate how IL-4, a key asthma-associated cytokine, can influence ADMA-related effects on lungs. Methods BEAS2B (bronchial epithelial) cells were treated with IL-4 followed by ADMA and investigated for oxo-nitrative stress and resultant mitochondrial toxicity after 48 hours by using flow cytometry, confocal imaging, immunoblotting, and fluorimetric assays. Results IL-4–induced mitotoxicity in BEAS2B cells was significantly higher in the presence of exogenous ADMA. IL-4 treatment led to proteolytic degradation of dimethylarginine dimethylaminohydrolase 2, which catabolizes ADMA. IL-4 pretreatment was associated with increased intracellular ADMA accumulation and increased ADMA-induced mitotoxicity. Airway epithelial cells treated with IL-4 followed by ADMA showed exaggerated oxo-nitrative stress and potent induction of the cellular hypoxic response, despite normoxic conditions. The hypoxic response was associated with reduced mitochondrial function but was reversible by overexpression of the mitochondrial biogenesis factor, mitochondrial transcription factor A. Conclusion We conclude that IL-4 promotes intracellular ADMA accumulation, leading to mitochondrial loss through oxo-nitrative stress and hypoxic response. This provides a novel understanding of how obesity, with high ADMA levels, and asthma, with high IL-4 levels, might potentiate each other and highlights the potential of mitochondrial-targeted therapeutics in obese subjects with asthma.