一种肝组织切片胶原定量分析法的实验性研究

Experimental study of a micromethod for collagen determination in hepatic tissue sections

目的 探讨肝纤维化组织切片上胶原测定方法。方法 用二甲基亚硝氨（ＤＭＮ） 诱导ＳＤ大鼠肝纤维化模型，常规肝组织切片，ＨＥ、Ｍａｓｓｏｎ 三色和苦味酸天狼红染色，苦味酸固绿天狼红染色比色测定及羟脯氨酸（Ｈｙｐ）含量测定。结果 对照组和肝纤维化明显组每张切片胶原含量分别为０．２３μｇ和１．０μｇ，非胶原蛋白含量分别为０．０６１ｍｇ 和０ ．０７１ｍｇ；切片胶原测定分别为３７ ．２３μｇ／ｍｇ·ｐｒｏ 和１２３．７μｇ／ｍｇ·ｐｒｏ；切片胶原与肝纤维化程度变化相一致，与肝组织Ｈｙｐ 含量具有良好的相关性，ｒ 均为０．９４ ，Ｐ＜０ ．０１ 。结论 切片胶原测定是一种简便、准确、可靠的肝纤维化程度的量化诊断方法。

Objective To search for a micromethod for quantitative collagen determination in tissue sections with liver fibrosis. Methods Dimethylnitrosamine (DMN) was used to produce hepatic fibrosis in SD rat model. Liver sections were stained with HE stain, Masson's trichome stain and Picric acid Sirus red stain. Collagen in liver sections was tested with Picric acid Sirus red and Fast green stain colorimetric technique, and liver hydroxyproline (Hyp) content was also determined.Results In sections of control and liver fibrosis group, collagen contents were 0.23μg and 1.0μg, respectively; non collagenes protein were 0.061mg and 0.071mg, respectively; collagen contents in the sections were 37.23μg/mg protein and 123.7μg/mg protein, respectively. The collagen content in sections was in consistency with the degree of hepatic fibrosis, and there was better correlation between Hyp content and collagen content in these sections. Their coefficients “r” were 0.94 ( P< 0.01).Conclusion These results suggest that the micromethod for collagen determination in tissue sections is a simple, accurate and reliable method for quantitative diagnostic indices in hepatic fibrosis.