Dunn M A, Rojkind M, Warren K S, Hait P K, Rifas L, Seifter S
J Clin Invest. 1977 Apr;59(4):666-74. doi: 10.1172/JCI108685.
Collagen synthesis was measured in liver slices obtained from mice with hepatosplenic schistosomiasis. Enlarged fibrotic livers from these mice contained 20 times more collagen than normal. This model of hepatic fibrosis results from an inflammatory granulomatous host response to Schistosoma mansoni ova in portal tracts, rather than from direct lover cell injury as with carbon tetrachloride-induced liver fibrosis. Collagen synthesis, as measured by the formation of labeled protein-bound hydroxyproline, occurred in granulomas isolated from fibrotic livers. Labeled collagen that cochromatographed with type I collagen was extracted with neutral salt solution from liver slices incubated with labeled proline. The free proline pool of the liver was doubled in infected mice; coordinately, liver slices from these animals showed maximal collagen production when the concentration of free proline in the medium was raised to 0.4 mM, the same level measured in the fibrotic livers. Under such conditions, collagen synthesis was at a rate equivalent to the formation of 5.4 nmol of protein-bound hydroxyproline per g liver in 6 h. In comparative incubations in medium containing 0.2 mM proline, fibrotic liver slices produced 16-fold more collagen than normal slices. The proline analogue, L-azetidine 2-carboxylic acid, effectively inhibited synthesis of labeled collagen by fibrotic liver slices. These studies show the synthesis of collagen in a reproducible animal model of the most prevalent form of human liver fibrosis. Difinitition of the controlling factors in this system is of interest for the general problem of fibrosis produced by immunological responses.
在患有肝脾血吸虫病的小鼠所获取的肝切片中测量了胶原蛋白的合成。这些小鼠肿大的纤维化肝脏所含的胶原蛋白比正常肝脏多20倍。这种肝纤维化模型是由宿主对门脉区曼氏血吸虫卵的炎性肉芽肿反应所致,而非像四氯化碳诱导的肝纤维化那样由直接的肝细胞损伤引起。通过标记的蛋白质结合羟脯氨酸的形成来测定,胶原蛋白合成发生在从纤维化肝脏分离出的肉芽肿中。与I型胶原共层析的标记胶原蛋白是用中性盐溶液从与标记脯氨酸一起孵育的肝切片中提取的。感染小鼠肝脏的游离脯氨酸池增加了一倍;相应地,当培养基中游离脯氨酸浓度提高到0.4 mM(纤维化肝脏中测得的相同水平)时,这些动物的肝切片显示出最大的胶原蛋白生成量。在这种条件下,胶原蛋白合成速率相当于每克肝脏在6小时内形成5.4 nmol蛋白质结合羟脯氨酸。在含有0.2 mM脯氨酸的培养基中进行的对比孵育中,纤维化肝切片产生的胶原蛋白比正常切片多16倍。脯氨酸类似物L-氮杂环丁烷2-羧酸有效地抑制了纤维化肝切片中标记胶原蛋白的合成。这些研究表明了在人类最常见的肝纤维化形式的可重复动物模型中胶原蛋白的合成。确定该系统中的控制因素对于免疫反应产生的纤维化这一普遍问题具有重要意义。