Proteome analysis of cardiomyocyte proteins in forensic autopsy cases and application to forensic pathological diagnosis

法医尸检案件心肌细胞蛋白的蛋白质组分析及其在法医病理诊断中的应用

• 批准号: 17590580
• 负责人: TSUDA Ryouichi
• 金额: $ 2.3万
• 依托单位: Nagasaki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17590580/
• 关键词: biomolecular; protein; cardiomyocyte; two-dimensional gel electrophoresis; proteome; forensic molecular pathology; forensic pathological diagnosis; social medicine; α-エノラーゼ; 二次元電気泳動; 法病理診断; MnSOD

We performed a differential proteomic profiling study on expression changes of cardiomyocyte proteins ; desmin, troponin T, gelsolin, α-smooth muscle actin (α-SMA), manganese superoxide dismutase (MnSOD) and α-enolase in left ventricular muscles of forensic autopsy cases by two-dimensional gel electrophoresis.1.Desmin : Des min abnormalities were reported in cardiomyopathy and heart failure cases. The significant differences were found between asphyxia and sudden cardiac death (p<0.03), and head injury (p<0.02), between bleeding and head injury (p<0.05) in the ratio of phosphorylated to dephosphorylated desmin.2. Gelsolin : Gelsolin is an abundant actin binding protein and the protein expression was increased in cardiomyopathic case. We demonstrate that isoelectric point (pI) of several gelsolin spots shifted to acidic region in 16 cases.3. Troponin T: Cardiac troponin T is an essential component of the cardiac contraction regulations and the sensitivity of cardiac myofilaments to calcium. We detected altered troponin T spots in addition to original spots in 5. In 6 cases, we couldn't find the tropnin T spots.4. α-SMA : In experimental rat models, hypertrophic stimuli induce early expression of 'fetal genes' including α-SKA and α-SMA. The expression of α-SMA in 51 forensic autopsy cases was divided into 3 groups : 1. Strong (n=9), 2. Moderate (n=21), 3. Weak or Negative (n=21).5. MnSOD : MnSOD is one of the main antioxidant enzymes that protect the heart against ischemia-reperfusion injury. In 19 cases, a new MnSOD spot was detected in basic region.6. α-Enolase : α-Enolase is a glycolytic enzyme and the enzyme expression is observed in rat heart subjected ischemia-reperfusion injury. We demonstrated over a two-fold overexpression of α-enolase in 19 cases including four sudden cardiac death cases.

我们用双向凝胶电泳法对法医尸检病例左室肌中结蛋白、肌钙蛋白T、明胶蛋白、α-平滑肌肌动蛋白(α-SMA)、锰超氧化物歧化酶(MnSOD)和α-烯醇化酶的表达变化进行了差异蛋白质组学研究。1.Desmin：在心肌病和心力衰竭病例中发现Des-min异常。结果：1.窒息与心源性猝死(p&lt；0.03)、脑损伤(p&lt；0.02)、出血与脑损伤(p&lt；0.05)之间的磷酸化/去磷酸化脱氢酶比值差异有统计学意义。明胶蛋白：明胶蛋白是一种丰富的肌动蛋白结合蛋白，在心肌病病例中表达增加。我们证明了16例中几个明胶斑点的等电点(等电点)向酸性区域移动。肌钙蛋白T：心肌肌钙蛋白T是心脏收缩调节的重要组成部分，也是心肌肌丝对钙的敏感性。除原有斑点外，5例发现肌钙蛋白T斑点改变。6例未发现肌钙蛋白T斑点。α-SMA：在实验大鼠模型中，肥大刺激诱导包括α-SKA和α-SMA在内的‘胎儿基因’的早期表达。将51例法医尸检标本中α-SMA的表达分为3组：1.强阳性(n=9)，2.中度(n=21)，3.弱或阴性(n=21)。MnSOD：MnSOD是保护心脏免受缺血再灌注损伤的主要抗氧化酶之一。在19例患者中，在碱性区域检测到一个新的MnSOD斑点。α-Enolase：α-Enolase是一种糖酵解酶，在大鼠心脏缺血再灌注损伤中观察到该酶的表达。在19例患者中，包括4例心脏性猝死患者，α-enolase的过度表达超过两倍。