【摘要】 分别用兔抗大鼠L型或K型丙酮酸激酶(PyK)的抗体为第一抗体,并以异硫氰酸荧光素(FITC)标记的羊抗兔IgG为第二抗体,用双抗体法对正常大鼠肝、肾切片及小鼠HepA腹水肝癌(简称HepA)细胞涂片作免疫荧光染色,借以探讨L型及K型PyK在这些组织中的定位。结果发现:(1)L型PyK主要存在于肝实质细胞,而K型PyK可能主要存在于柯否细胞。(2)HepA细胞的胞浆,被K型PyK抗体染色后呈强烈的荧光。(3)L型PyK仅存在于大鼠肾脏皮质中,而K型PyK在大鼠肾皮质和髓质中均较丰富。以上两型PyK在肾脏中的分布与皮质和髓质的糖代谢特性不同是相一致的。因肾髓质较皮质含有较多的K型PyK,故其氧化率低于皮质,而酵解率和Crabtree效应高于皮质。更多还原

【Abstract】 By using rabbit antibody against purified rat liver type L or type K pyruvate kinase (PyK) as the first antibody, and fluoresoein isothiocyanate-labelled sheep IgG against rabbit antibody as the second antibody, the localization of the type L and the type K PyK in normal rat liver, kidney and mouse Hep A ascites hepatoma (Hep A) has been studied by double antibody immunofluorescence method. It has been revealed that: (1) Type L PyK is located in parenohymal cells predominently, and type K PyK is mainly in Kupffer’s cells. (2) Hep A cells give strong fluorescence when stained by anti-type K PyK antibody. (3) In rat kidney, type L PyK is present in the cortex only, whereas type K PyK is located in both the cortex and the medulla in significant amount. The localization of the two types of PyK are in correlation with the difference of carbohydrate metabolism in the kidney cortex and medulla. The medulla contains more type K PyK than the cortex, so the oxidation rate in the medulla homogenate is lower than that in the cortex, whereas the glycolytio rate and the Orabtree effect are higher.更多还原