CELLULAR PATHOPHYSIOLOGY OF HEPATIC FIBROSIS

肝脏含有一种细胞外基质， 窦周（Disse）间隙，成分与基底层相似 一般是上皮组织。 原发性肝癌肝细胞的研究进展 文化表明，基质，或其组成蛋白质，发挥主要作用， 在调节肝脏特异性功能表达中的作用。 肝细胞 似乎以饱和的方式与基质蛋白相互作用，表明 特定受体的存在。 在本申请中，我们建议 分离基底膜主要蛋白质的肝细胞表面受体， 层（纤连蛋白，胶原IV和层粘连蛋白），使用最近描述的 将培养的肝细胞上的受体原位交联到 基质蛋白作为培养底物。 交联是 通过可裂解的光活化的异双官能的 试剂4 ′-叠氮基偶氮苯-4-氧代琥珀酰亚胺酯。 初步研究由 主要研究者已经证明， 不同的分子大小，可能代表个体的受体 基质蛋白可以通过这种方法分离。 抗体（两者 多价和单克隆）将被提高到这些蛋白质， 表征它们并操纵细胞-基质相互作用， 文化 它们也将用于开发受体的免疫测定法。 肝脏中的浓度。 存在肝细胞癌 伴随肝脏炎症的功能障碍不仅反映了 Disse间隙中的异常成分，但也改变了 基质受体 这些实验预计将提供详细的新的 细胞-基质相互作用及其对肝特异性 功能协调发展的

The liver contains an extracellular matrix that is localized to the perisinusoidal (Disse) space and similar in composition to the basal lamina of epithelial tissues generally. Recent studies of hepatocytes in primary culture suggest that the matrix, or its constituent proteins, play a major role in modulating the expression of liver-specific functions. Hepatocytes appear to interact with matrix proteins in a saturable manner, suggesting the presence of specific receptors. In this application, we propose to isolate hepatocyte surface receptors for the major proteins of the basal lamina (fibronectin, collagen IV and laminin) using a recently described method for in situ cross-linking of receptors on cultured hepatocytes to the matrix protein serving as culture substratum. The cross-linking is accomplished by means of the cleavable photoactivable heterobifunctional reagent 4'-azidoazobenzene-4-oxysuccinimide ester. Preliminary studies by the Principal Investigator have demonstrated that hepatocyte proteins of distinct molecular size, potentially representing receptors for individual matrix proteins, can be isolated by this approach. Antibodies (both polyvalent and monoclonal) will be raised to these proteins, both for characterizing them and for manipulating cell-matrix interaction in culture. They will be used also to develop an immunoassay for receptor concentration in the liver. The possibility exists that hepatocellular dysfunction accompanying hepatic inflammation reflects not only a matrix of abnormal composition in the Disse space but also altered expression of matrix receptors. The experiments are expected to provide detailed new insight into cell-matrix interaction and its regulation of liver-specific functions.