James J, Bosch K S, Aronson D C, Houtkooper J M
Laboratory of Histology and Cell Biology, University of Amsterdam, The Netherlands.
Liver. 1990 Feb;10(1):1-5. doi: 10.1111/j.1600-0676.1990.tb00428.x.
By means of staining with Sirius Red F3BA in a saturated picric acid solution, the collagen contents of rat livers with varying degrees of fibrosis have been measured quantitatively in fixed and sectioned material, using both histophotometry in situ and extraction of bound dye with colorimetric analysis. These findings have been correlated with chemical assays of the hydroxyproline content in homogenates from the same livers. It appears that a highly significant correlation exists between both section-based analysis methods and the hydroxyproline content, the Spearman-Rank correlation coefficients being virtually identical. For analysis of collagen accumulation in rat liver, both section-based methods seem to be useful and reliable, the extraction method giving the quickest results for large-scale screening, and the histophotometric method being more appropriate to take readings in selected areas. With human liver material, indications have been obtained for the existence of large sampling errors due to inhomogeneous distribution of collagen deposits. Using the extraction method, no significant changes could be observed in the volume density of collagen during postnatal growth from 1 week to 21 months in rat liver: only on the third day after birth was a higher value of collagen/total protein obtained, possibly due to a higher water content of the hepatocytes. Partial hepatectomy was found to have no influence at all on the collagen content of rat liver during the period of restorative growth or after it.
通过在饱和苦味酸溶液中用天狼星红F3BA染色,对不同程度纤维化的大鼠肝脏固定切片材料中的胶原蛋白含量进行了定量测定,采用原位组织光度测定法和比色分析法提取结合染料。这些发现与来自同一肝脏匀浆中羟脯氨酸含量的化学测定结果相关。似乎基于切片的两种分析方法与羟脯氨酸含量之间都存在高度显著的相关性,斯皮尔曼等级相关系数实际上是相同的。对于大鼠肝脏中胶原蛋白积累的分析,基于切片的两种方法似乎都是有用且可靠的,提取法对于大规模筛选能给出最快的结果,而组织光度测定法更适合在选定区域进行读数。对于人类肝脏材料,由于胶原蛋白沉积分布不均,已发现存在较大的抽样误差迹象。采用提取法,在大鼠肝脏从出生1周到21个月的产后生长过程中,未观察到胶原蛋白的体积密度有显著变化:仅在出生后第三天获得了较高的胶原蛋白/总蛋白值,这可能是由于肝细胞含水量较高。发现部分肝切除在恢复性生长期间或之后对大鼠肝脏的胶原蛋白含量完全没有影响。