【摘要】 用破碎、酶解、盐析三步法提纯大鼠肝组织Ⅰ型胶原蛋白,并用 ECL-Western blot测定正常及纤维化肝组织中Ⅰ型胶原蛋白的含量,来建立一种有效、简便的大鼠肝组织Ⅰ型胶原蛋白的提取、纯化及鉴定的方法,研究抗纤维化药物的作用机制。结果发现,该法提取Ⅰ型胶原蛋白的时间周期短,得率高,纯度达到PAGE纯的水平,并且检测灵敏度高。大鼠轻度肝纤维化时,即可发现Ⅰ型胶原蛋白含量明显增多,且可被抗纤维化药物逆转。结果表明该法具有简便、灵敏和无放射性污染等优点,为研究肝纤维化机制及筛选抗纤维化药物提供了有价值的方法。更多还原

【Abstract】 In order to establish a simple and effective method to extract, purify and detect type Ⅰ collagen fromrat livers, and study the mechanisms of drugs on the hepatic fibrosis, the liver fibrotic models were induced withpig serum in rats, and treated with colchicine. Type Ⅰ collagen in rat livers was extracted through three steps:crushing, acidolysing and salting out. The contents of type Ⅰ collagen from normal and fibrofic livers weredetected by SDS-PAGE and Western blot to determine its variety in hepatic fibrosis, at the same time the liverhistopathological changes were also evaluated. Results showed that the method could shorten the extractedduration and enhance the output rate, and the obtained collagen was proved to have high electrophoretic purity.It was also found that the content of type Ⅰ collagen in liver increased evidently during the hepatic fibrosis, andthe antifibrosis drug could reverse it. So we can draw a conclusion that a simple, rapid, sensitive and non-radioative method is established successfully. The method also provides a valuable means for studying thepathological mechanism of hepatic fibrosis, and screening antifibrosis drugs.更多还原