实验性阿霉素心肌炎心肌肌钙蛋白Ⅰ免疫组化研究

An immunohistochemical study with cardiac troponin I in experimental adriamycin myocarditis

心肌肌钙蛋白I(cTnI)是诊断心肌损伤敏感特异的生化指标 ,已广泛应用于缺血性心肌损伤的诊断 ,对于终末期心力衰竭中cTnI的改变也做了较为深入研究 ,但关于心肌炎中cTnI的研究尚很少。目的 应用抗cTnI单克隆抗体对实验性心肌炎进行免疫组化研究 ,观察其在炎症心肌中的定位分布 ,初步探讨cTnI在心肌炎评估、诊断中的重要意义 ,为cTnI应用于心肌炎诊断提供重要实验和理论依据。方法 提取纯化人cTnI,制备抗cTnI单克隆抗体和SD大鼠心肌炎模型 ,对正常和心肌炎大鼠心肌行HE和SP染色。结果 HE染色 :正常大鼠心肌细胞呈梭形 ,核清蓝色 ,肌纤维呈深红色 ,胞浆淡红色。心肌炎大鼠心肌散在局灶病变区 ,表现为两种不同的变化 ,一是心肌细胞变性伴不同程度的心肌坏死 ,二是肌纤维溶解坏死。SP染色 :正常心肌细胞不着色。炎症区心肌细胞着色变深呈不均匀细颗粒状。坏死区心肌细胞和肌纤维均不着色 ,仅余网状细胞骨架呈棕褐色着色。结论 应用cTnI能较好地反映心肌炎症的部位、程度 ,结合外周血cTnI水平的测定 。

Background Cardiac troponin Ⅰ(cTnI), as a sensitive and specific biochemical mark of cardiac injuries, has been widely used in diagnosing of myocadial ischemia, recently its alteration in end stage heart failure has also been recognized profoundly, but it was known fewer in myocarditis.Objective\ This study was designed to determine the inflammatory region and degree in experimental myocarditis by immunohistochemistry, and to assess the test performance characteristics of histionic cardiac troponin Ⅰ (cTnI) for diagnosis of myocarditis.Methods Balb/c mice were immunized with cTnI. Several high specific monoclonal anti-cTnI antibodies were developed after cell fusion, cloning and screening. The localization and degree of myocardial inflammation were analyzed by HE and SP staining.Results The 3D2 were selected as the first antibody among the five hydridoma cell lines. After SP staining, the area of inflammatory myocardium displayed as an uneven granular material, the reticulate cell frameworks in the necrosis area were dyed brown, while the normal myocardium were not be stained.Conclusion The specific monoclonal anti-cTnI antibody could reveal the localization and degree of the inflammation in myocarditis, and could be used as a diagnostic method to myocarditis.