Bristol Heart Institute and Bristol Medical School, University of Bristol, Bristol BS2 8HW, UK.
National Heart and Lung Institute, Imperial College, London SW3 6LY, UK.
Int J Mol Med. 2022 Jun;49(6). doi: 10.3892/ijmm.2022.5133. Epub 2022 Apr 15.
Our earlier work has shown inter‑disease and intra‑disease differences in the cardiac proteome between right (RV) and left (LV) ventricles of patients with aortic valve stenosis (AVS) or coronary artery disease (CAD). Whether disease remodeling also affects acute changes occuring in the proteome during surgical intervention is unknown. This study investigated the effects of cardioplegic arrest on cardiac proteins/phosphoproteins in LV and RV of CAD (n=6) and AVS (n=6) patients undergoing cardiac surgery. LV and RV biopsies were collected during surgery before ischemic cold blood cardioplegic arrest (pre) and 20 min after reperfusion (post). Tissues were snap frozen, proteins extracted, and the extracts were used for proteomic and phosphoproteomic analysis using Tandem Mass Tag (TMT) analysis. The results were analysed using QuickGO and Ingenuity Pathway Analysis softwares. For each comparision, our proteomic analysis identified more than 3,000 proteins which could be detected in both the pre and Post samples. Cardioplegic arrest and reperfusion were associated with significant differential expression of 24 (LV) and 120 (RV) proteins in the CAD patients, which were linked to mitochondrial function, inflammation and cardiac contraction. By contrast, AVS patients showed differential expression of only 3 LV proteins and 2 RV proteins, despite a significantly longer duration of ischaemic cardioplegic arrest. The relative expression of 41 phosphoproteins was significantly altered in CAD patients, with 18 phosphoproteins showing altered expression in AVS patients. Inflammatory pathways were implicated in the changes in phosphoprotein expression in both groups. Inter‑disease comparison for the same ventricular chamber at both timepoints revealed differences relating to inflammation and adrenergic and calcium signalling. In conclusion, the present study found that ischemic arrest and reperfusion trigger different changes in the proteomes and phosphoproteomes of LV and RV of CAD and AVS patients undergoing surgery, with markedly more changes in CAD patients despite a significantly shorter ischaemic period.
我们之前的研究表明，在主动脉瓣狭窄（AVS）或冠状动脉疾病（CAD）患者的右心室（RV）和左心室（LV）之间，心脏蛋白质组存在疾病间和疾病内差异。尚不清楚疾病重塑是否也会影响手术干预期间蛋白质组中发生的急性变化。本研究调查了心脏停搏对接受心脏手术的 CAD（n=6）和 AVS（n=6）患者 LV 和 RV 心脏蛋白/磷酸蛋白的影响。在缺血冷血心脏停搏之前（术前）和再灌注后 20 分钟（术后）收集手术期间的 LV 和 RV 活检。组织立即冷冻，提取蛋白质，并用串联质量标签（TMT）分析提取的提取物进行蛋白质组学和磷酸蛋白质组学分析。使用 QuickGO 和 Ingenuity Pathway Analysis 软件分析结果。对于每个比较，我们的蛋白质组学分析在术前和术后样本中均检测到超过 3000 种蛋白质。心脏停搏和再灌注与 CAD 患者 LV 中的 24 个（LV）和 120 个（RV）蛋白质的显著差异表达相关，这些蛋白质与线粒体功能、炎症和心脏收缩有关。相比之下，尽管缺血性心脏停搏时间明显更长，但 AVS 患者仅显示 3 个 LV 蛋白和 2 个 RV 蛋白的差异表达。CAD 患者中有 41 个磷酸蛋白的相对表达发生显著改变，而 AVS 患者中有 18 个磷酸蛋白的表达发生改变。炎症途径与两组磷酸蛋白表达的变化有关。在同一心室室的两种疾病比较中，发现与炎症和肾上腺素能和钙信号有关的差异。总之，本研究发现，缺血性停搏和再灌注在手术中 CAD 和 AVS 患者的 LV 和 RV 蛋白质组和磷酸蛋白质组中引发不同的变化，尽管 CAD 患者的缺血期明显缩短，但变化明显更多。
