Hossain Mohammad A, De Souza Ayesha I, Bagul Atul, MacPhee Iain A M, Kessaris Nicos, Morsy Mohamed A
Department of Renal Transplantation, St. George's Hospital NHS Trust, Blackshaw Rd., Tooting, London SW17 0QT, United Kingdom.
Cardiovascular Sciences Research Centre, Division of Clinical Sciences, St. George's, University of London, Cranmer Terrace, Tooting, London SW17 0RE, United Kingdom.
J Proteomics. 2014 Aug 28;108:133-45. doi: 10.1016/j.jprot.2014.05.008. Epub 2014 May 24.
The use of donation after circulatory death (DCD) kidneys for transplantation is increasing. Subsequent delayed graft function is related to ischaemia/reperfusion injury (I/R), warm ischaemia (WI) being one of the main contributing factors. This proteomics study aimed to identify candidate biomarkers of WI.
Termination biopsies were obtained over 180min in 6 pigs. Proteins were subjected to differential in-gel electrophoresis (DIGE) and identified using LC MS/MS.
Thirty nine protein spots showed significant changes in expression (ANOVA, p<0.05). Peroxiredoxin-3 and -6 (PRX3 and PRX6) were expressed with a fold change (FD) of +1.8 (p=0.03 and 0.02 respectively). A significant upregulation of Alpha-2-HS-glycoprotein (A2HSG, FD+1.9, p=0.047) and heat-shock protein 70-1b (HSP70-1b, FD+2.1 p=0.002) was recorded.
The expression of PRX3, PRX6 and HSP70-1b during the first 30min of WI may be critical in measuring cellular responses. This is the first large animal model to describe the novel candidate biomarker, structural protein A2HSG. A2HSG upregulation during WI alone in this study is encouraging and further assessment in a DCD auto-transplant model is warranted.
Warm ischaemia (WI) during donation after circulatory death (DCD) organ retrieval is associated with higher rates of post transplant organ dysfunction. The cellular and molecular mechanism of this paradigm is poorly reported. The work carried out in this large animal study has been performed to enable better understanding of protein expression during DCD WI at the time of retrieval. We have identified differential increased expression of PRX3, PRX6 and HSP70 during the first 30min of WI. Observation of this behaviour has not been reported before. Application of these results in a reperfusion model or autograft animal study would further help study of the named proteins as clinical biomarkers of WI. Alpha 2-HS Glycoprotein (A2HSG) species were also differentially expressed during the WI period. This remains a novel finding. Assessment of A2HSG is also recommended for further study in a reperfusion context. Previous reports of A2HSG have suggested an association in chronic kidney disease and diabetes, but no association with WI has previously been noted in either small or large animals.
用于移植的心脏死亡后捐赠（DCD）肾脏的使用正在增加。随后的移植肾功能延迟与缺血/再灌注损伤（I/R）有关，热缺血（WI）是主要促成因素之一。这项蛋白质组学研究旨在识别WI的候选生物标志物。
在6头猪身上于180分钟内获取终止活检组织。对蛋白质进行差异凝胶内电泳（DIGE），并使用液相色谱串联质谱（LC MS/MS）进行鉴定。
39个蛋白点显示出表达的显著变化（方差分析，p<0.05）。过氧化物酶体增殖物激活受体3和6（PRX3和PRX6）的表达倍数变化（FD）为+1.8（分别为p=0.03和0.02）。记录到α2-HS糖蛋白（A2HSG，FD+1.9，p=0.047）和热休克蛋白70-1b（HSP70-1b，FD+2.1，p=0.002）显著上调。
WI最初30分钟内PRX3、PRX6和HSP70-1b的表达可能对测量细胞反应至关重要。这是第一个描述新型候选生物标志物结构蛋白A2HSG的大型动物模型。本研究中仅WI期间A2HSG的上调令人鼓舞，有必要在DCD自体移植模型中进行进一步评估。
心脏死亡后捐赠（DCD）器官获取过程中的热缺血（WI）与移植后器官功能障碍的较高发生率相关。这种模式的细胞和分子机制报道较少。在这项大型动物研究中开展的工作是为了更好地理解DCD WI获取时的蛋白质表达。我们已经确定WI最初30分钟内PRX3、PRX6和HSP70表达差异增加。此前尚未报道过这种行为。将这些结果应用于再灌注模型或自体移植动物研究将进一步有助于将所命名的蛋白质作为WI的临床生物标志物进行研究。α2-HS糖蛋白（A2HSG）在WI期间也有差异表达。这仍然是一个新发现。也建议在再灌注背景下对A2HSG进行进一步研究。此前关于A2HSG的报道表明其与慢性肾病和糖尿病有关，但在小型或大型动物中此前均未发现其与WI有关。
