Riezzo Irene, Cantatore Santina, DeCarlo Dania, Fiore Carmela, Neri Margherita, Turillazzi Emanuela, Fineschi Vittorio
Department of Forensic Pathology, Ospedale Colonnello D'Avanzo, Viale degli Aviatori 1, 71100 Foggia, Italy.
Curr Vasc Pharmacol. 2015;13(1):78-90. doi: 10.2174/15701611113119990004.
Cardiac muscle necrosis is associated with inflammatory cascade that clears the infarct from dead cells and matrix debris, and then replaces the damaged tissue with scar, through three overlapping phases: the inflammatory phase, the proliferative phase and the maturation phase. Western blotting, laser confocal microscopy, Raman microscopy are valuable tools for studying the inflammatory response following myocardial infarction both humoral and cellular phase, allowing the identification and semiquantitative analysis of proteins produced during the inflammatory cascade activation and the topographical distribution and expression of proteins and cells involved in myocardial inflammation. Confocal laser scanning microscopy (CLSM) is a relatively new technique for microscopic imaging, that allows greater resolution, optical sectioning of the sample and three-dimensional reconstruction of the same sample. Western blotting used to detect the presence of a specific protein with antibody-antigen interaction in the midst of a complex protein mixture extracted from cells, produced semi-quantitative data quite easy to interpret. Confocal Raman microscopy combines the three-dimensional optical resolution of confocal microscopy and the sensitivity to molecular vibrations, which characterizes Raman spectroscopy. The combined use of western blotting and confocal microscope allows detecting the presence of proteins in the sample and trying to observe the exact location within the tissue, or the topographical distribution of the same. Once demonstrated the presence of proteins (cytokines, chemokines, etc.) is important to know the topographical distribution, obtaining in this way additional information regarding the extension of the inflammatory process in function of the time stayed from the time of myocardial infarction. These methods may be useful to study and define the expression of a wide range of inflammatory mediators at several different timepoints providing a more detailed analysis of the time course of the infarct.
心肌坏死与炎症级联反应相关，该反应清除梗死区域的死亡细胞和基质碎片，然后通过三个重叠阶段用瘢痕替代受损组织：炎症期、增殖期和成熟期。蛋白质免疫印迹法、激光共聚焦显微镜、拉曼显微镜是研究心肌梗死后炎症反应（包括体液和细胞阶段）的重要工具，可用于鉴定和半定量分析炎症级联激活过程中产生的蛋白质，以及心肌炎症中涉及的蛋白质和细胞的拓扑分布及表达。共聚焦激光扫描显微镜（CLSM）是一种相对较新的显微成像技术，具有更高的分辨率，能够对样品进行光学切片和对同一样品进行三维重建。蛋白质免疫印迹法用于在从细胞中提取的复杂蛋白质混合物中通过抗体 - 抗原相互作用检测特定蛋白质的存在，产生的半定量数据易于解释。共聚焦拉曼显微镜结合了共聚焦显微镜的三维光学分辨率和拉曼光谱对分子振动的敏感性。蛋白质免疫印迹法和共聚焦显微镜的联合使用可以检测样品中蛋白质的存在，并试图观察其在组织内的确切位置或相同蛋白质的拓扑分布。一旦证明蛋白质（细胞因子、趋化因子等）的存在，了解其拓扑分布就很重要，这样可以获得关于心肌梗死后炎症过程随时间延长的额外信息。这些方法可能有助于研究和定义多种炎症介质在几个不同时间点的表达，从而更详细地分析梗死的时间进程。
