Histochemical Microquantitation for Extracellular Matrix Proteins and mRNAs in Mammalian Histologic Sections

冰冻组织切片中的细胞外基质(ECM)蛋白和mRNAs用本研究项目发展的组织化学微量方法进行了定量。例如，测定了实验性肾小球肾炎大鼠肾皮质和病毒性肝炎患者肝脏中ECM的含量。ECM含量的增加与肾脏疾病的常规生化指标有很好的相关性。免疫组织化学显微方法可定量评价系膜增生性肾小球肾炎的组织病理学诊断。这一新的组织化学微量定量技术可用于其他生物物质的定量，这些生物物质在细胞生理功能中发挥着重要作用。例如，冰冻组织切片中的快速肌球蛋白通过组织化学微量方法进行定量。用显微技术对哺乳动物骨骼肌中的快速肌球蛋白水平进行了定量。总之，该微量方法具有足够的灵敏度，可以检测小切片中ECM蛋白和mRNAs的含量，并使定量检测活检切片中的ECM水平成为可能。该微量方法可用于疾病的组织病理学诊断，精确评估器官的生理和病理特征，以及评估药物的疗效。

Extracellular matrix (ECM) proteins and mRNAs in frozen histologic sections were quantified by a histochemical microquantitation methods, which are developed in the present research project. For instance, ECM contents in renal cortex of rats with experimental glomerulonephritis and in liver from the patients with viral hepatitis were measured. The increase in ECM contents correlated well with conventional biochemical parameters for renal disorders. The immunohistochemical micromethod may make it possible to evaluate the histopathological diagnosis of mesangial proliferative glomerulonephritis quantitatively. This novel histochemical microquantitation technique is able to use for quantification of other biological substances, which plays an important role in the cellular physiological functions. For example, fast-myosin in frozen histological sections was quantified by a histochemical micromethod. Fast-myosin levels in mammalian skeletall muscles were quantified by the microtechnique. In conclusion, the micromethod is sensitive enough to measure the content of ECM proteins and mRNAs in the small sections, and make it possible to quantify the ECM levels in the biopsy sections. The micromethod is useful for histopathological diagnosis of the diseases, for precise evaluation of physiological and pathological characteristics of the organs, and for estimation of therapeutic efficacy of drugs.