title: Induction of IL-8(CXCL8) and MCP-1(CCL2) with oxidative stress and its inhibition with N-acetyl cysteine (NAC) in cell culture model using HK-2 cell.
abstract: Renal transplantation can often be complicated due to delayed graft function, which is a direct sequel of ischaemia reperfusion injury. The adverse outcome of delayed graft function is not only short term but the long-term function of the graft is also affected. Therefore, it is important to understand the mechanisms of ischaemia reperfusion injury. Reactive oxygen species are the key mediators in ischaemia reperfusion injury causing direct cell damage which also initiate inflammation by inducing chemokines. The presence of inflammation is a marker of severe delayed graft function. However, the effect of oxidative stress on the expression of key chemokines has not been fully established yet. Therefore, the aim of this study was to measure the oxidative stress response and the secretion of chemokines in a cell culture model that mimics the effects of ischaemia reperfusion injury in immortalised human renal proximal tubular epithelial cells, HK-2. Cells were treated with varying concentrations of hydrogen peroxide and markers of oxidative stress response and chemokine release were measured. Exposure to hydrogen peroxide induced a significant increase in the activity of the antioxidant enzyme glutathione peroxidase and the levels of the chemokines Interleukin-8 (IL-8; CXCL8) and MCP-1 (CCL2). A dose related increase of chemokine secretion was also observed. The cytokine Interleukin-1β (IL-1β) at 1 ng/ml significantly potentiated the expression of both IL-8 (CXCL8) and MCP-1 (CCL2) which showed synergistic response in the presence of hydrogen peroxide. Pre-incubation of the cells with the anti-oxidant N-acetyl cysteine (NAC) strongly suppressed the induction of both IL-8 and MCP-1 when stimulated with hydrogen peroxide and IL-1β. This study demonstrates the potential of anti-oxidants like N-acetyl cysteine in ameliorating the effects of ischaemia reperfusion injury thus suggesting a new therapeutic approach in renal transplantation. These findings can have potential implications for clinical use to prevent ischaemia reperfusion injury in renal transplantation.
authors.affiliation: Department of Musculoskeletal Biology, Institute of Ageing and Chronic Disease, University of Liverpool, UK; Peter Medawar Transplant Unit, Royal Liverpool University Hospital, Liverpool, UK.
authors.forename: Avneesh
authors.initials: A
authors.lastname: Kumar
authors.affiliation: Department of Musculoskeletal Biology, Institute of Ageing and Chronic Disease, University of Liverpool, UK.
authors.forename: Liliana
authors.initials: L
authors.lastname: Shalmanova
authors.affiliation: Peter Medawar Transplant Unit, Royal Liverpool University Hospital, Liverpool, UK.
authors.forename: Abdul
authors.initials: A
authors.lastname: Hammad
authors.affiliation: Department of Clinical Infection, Microbiology and Immunology, Institute of Infection & Global Health, University of Liverpool, UK.
authors.forename: Stephen E
authors.initials: SE
authors.lastname: Christmas
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology*; D018929:Cell Culture Techniques; D002460:Cell Line; D018932:Chemokine CCL2 / Q000276:immunology*; D006801:Humans; D053583:Interleukin-1beta / Q000276:immunology; D016209:Interleukin-8 / Q000276:immunology*; D016030:Kidney Transplantation; D018384:Oxidative Stress / Q000187:drug effects* / Q000276:immunology; D015427:Reperfusion Injury / Q000188:drug therapy* / Q000276:immunology
chemical_list: C111835:CCL2 protein, human; C508606:CXCL8 protein, human; D018932:Chemokine CCL2; C508585:IL1B protein, human; D053583:Interleukin-1beta; D016209:Interleukin-8; D000111:Acetylcysteine
keywords: CCL2); CXCL8); HK-2 (human kidney-2); Interleukin-8 (IL-8; Monocyte chemo attractant protein (MCP-1; N-acetyl cysteine (NAC); Oxidative stress