Kumar Avneesh, Shalmanova Liliana, Hammad Abdul, Christmas Stephen E
Department of Musculoskeletal Biology, Institute of Ageing and Chronic Disease, University of Liverpool, UK; Peter Medawar Transplant Unit, Royal Liverpool University Hospital, Liverpool, UK.
Department of Musculoskeletal Biology, Institute of Ageing and Chronic Disease, University of Liverpool, UK.
Transpl Immunol. 2016 Mar;35:40-6. doi: 10.1016/j.trim.2016.02.003. Epub 2016 Feb 22.
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.
肾移植常常会因移植肾功能延迟而变得复杂，移植肾功能延迟是缺血再灌注损伤的直接后果。移植肾功能延迟的不良后果不仅体现在短期，移植肾的长期功能也会受到影响。因此，了解缺血再灌注损伤的机制很重要。活性氧是缺血再灌注损伤中的关键介质，可导致直接的细胞损伤，还通过诱导趋化因子引发炎症。炎症的存在是严重移植肾功能延迟的一个标志。然而，氧化应激对关键趋化因子表达的影响尚未完全明确。因此，本研究的目的是在一个模拟缺血再灌注损伤效应的细胞培养模型中，检测永生化人肾近端小管上皮细胞HK-2的氧化应激反应和趋化因子分泌情况。用不同浓度的过氧化氢处理细胞，并检测氧化应激反应标志物和趋化因子释放情况。暴露于过氧化氢会导致抗氧化酶谷胱甘肽过氧化物酶的活性显著增加，以及趋化因子白细胞介素-8（IL-8；CXCL8）和单核细胞趋化蛋白-1（MCP-1；CCL2）水平显著升高。还观察到趋化因子分泌呈剂量相关增加。1 ng/ml的细胞因子白细胞介素-1β（IL-1β）显著增强了IL-8（CXCL8）和MCP-1（CCL2）的表达，在过氧化氢存在的情况下表现出协同反应。用抗氧化剂N-乙酰半胱氨酸（NAC）对细胞进行预孵育，在用过氧化氢和IL-1β刺激时，强烈抑制了IL-8和MCP-1的诱导。本研究证明了像N-乙酰半胱氨酸这样的抗氧化剂在减轻缺血再灌注损伤影响方面的潜力，从而为肾移植提出了一种新的治疗方法。这些发现可能对预防肾移植中缺血再灌注损伤的临床应用具有潜在意义。
