Klimek F, Moore M A, Schneider E, Bannasch P
Institute for Experimental Pathology, German Cancer Research Center, Heidelberg.
Histochemistry. 1988;90(1):37-42. doi: 10.1007/BF00495704.
A new method for the histochemical demonstration of pyruvate kinase (PK) activity was developed using a semi-permeable membrane and ATP-dependent phosphorylation of glucose coupled with tetrazolium reduction via glucose-6-phosphate dehydrogenase (G6PD) in order to investigate normal liver tissue and neoplastic hepatic nodules induced by thioacetamide (TAA). A series of control reactions and comparison with microbiochemical analysis of microdissected lyophilised material were used to determine the specificity of the reaction. In agreement with earlier reports, an activity gradient in control liver decreasing from zone 3 to zone 1 was apparent both histochemically and after biochemical analysis. Liver neoplastic nodules induced by 25 weeks dietary thioacetamide administration and characterized by increased G6PD demonstrated a clear decrease in PK activity. In contrast, epithelial cells within areas of cholangiocellular tumour development were characterized by a strong increase. Comparison of the results with immunohistochemical and biochemical data from the literature indicate that the specific histochemical method described will be of great assistance in future assessment of disease and physiological alteration in activity of this key enzyme of glycolysis.
为了研究正常肝组织以及硫代乙酰胺（TAA）诱导的肿瘤性肝结节，开发了一种新的组织化学方法来显示丙酮酸激酶（PK）活性，该方法使用半透膜以及葡萄糖的ATP依赖性磷酸化，并通过葡萄糖-6-磷酸脱氢酶（G6PD）偶联四氮唑还原反应。通过一系列对照反应以及与显微解剖冻干材料的微生物化学分析进行比较，以确定该反应的特异性。与早期报告一致，对照肝脏中从3区到1区的活性梯度在组织化学和生化分析后均很明显。经25周硫代乙酰胺饮食给药诱导并以G6PD增加为特征的肝脏肿瘤性结节显示PK活性明显降低。相反，胆管细胞肿瘤发展区域内的上皮细胞表现为强烈增加。将结果与文献中的免疫组织化学和生化数据进行比较表明，所描述的特定组织化学方法将对未来评估该糖酵解关键酶活性的疾病和生理改变有很大帮助。
