{
  "abstract": "Coronary artery disease (CAD) is a leading cause of global morbidity and mortality 1. In CAD, inflammation produces systolic and diastolic cardiac dysfunction. While cytokines and oxidative stress are well recognised pathogenic mediators, their interaction and contribution to cardiac cellular dysfunction remains unclear. Here, we determined the extent to which clinically relevant cytokines levels can elevate oxidative stress in cardiac myoblasts. We also began to elucidate the underlying sources of reactive oxygen species.In accordance with local and IRAS (ID: 247341) ethical approval, preoperative serum samples were collected from patients scheduled for coronary revascularisation surgery. Serum interleukin-6 (IL-6), -10 (IL-10), -1β (IL-1β) and tumour necrosis factor alpha (TNF-α) concentrations were measured using high-sensitivity ELISA (Abcam, UK & Invitrogen, USA). Cardiac myoblasts (H9c2) were incubated with these cytokines for 1 hour then loaded with CM-DCFDA for photometric measurement of oxidative stress.Average IL-6, IL-1β, IL-10 and TNF-α concentrations were 9.1 ± 1.36, 1.46 ± 0.26, 4.21 ± 0.59 and 2.70 ± 0.61 pg/mL, respectively. At these, and supra-pathological concentrations, only TNF-α and IL-6 produced a significant increase in DCFDA fluorescence. For instance, at a concentration of 1 ng/mL, TNF-α and IL-6 increased fluorescence by 71.67 ± 14.90% (n = 20, p = 0.0002) and 41.46 ± 12.49 (n = 20 , p = 0.0007) respectively. This effect was abolished by N-acetyl cysteine and attenuated by the NADPH oxidase inhibitor VAS3947. However, the xanthine oxidoreductase inhibitor Febuxostat only attenuated the effect in response to TNF-α.These data suggest that at clinically relevant concentrations, TNF-α and IL-6, but not IL-1β or IL-10 can increase oxidative stress in cardiac myoblasts. At least part of this effect is NADPH oxidase and xanthine oxidoreductase dependent. Our next experiments will determine whether these findings can be reproduced in iPSC-derived cells and probe the contribution of mitochondrial dysfunction.",
  "authors": [
    {
      "affiliations": [
        "University of Salford, Salford, United Kingdom"
      ],
      "name": "Bethan Samphire-Noden"
    },
    {
      "affiliations": [
        "University of Salford, Salford, United Kingdom"
      ],
      "name": "Alicia Staley"
    },
    {
      "affiliations": [
        "University of Salford, Salford, United Kingdom"
      ],
      "name": "Matthew Jones"
    },
    {
      "affiliations": [
        "University of Salford, Salford, United Kingdom"
      ],
      "name": "Sarah Withers"
    },
    {
      "affiliations": [
        "Blackpool Victoria Hospital, Blackpool, United Kingdom"
      ],
      "name": "Mohamad Nidal Bittar"
    },
    {
      "affiliations": [
        "University of Salford, Salford, United Kingdom"
      ],
      "name": "David Greensmith"
    }
  ],
  "title": "207 The cellular basis of coronary artery disease; interaction of cytokines and oxidative stress in cardiac myoblasts.",
  "uid": "ca34595f-885e-5e3a-8931-e6c84e749f30"
}