激光共聚焦显微镜对肝组织TGF-β_1的荧光定量分析

Quantitative analysis of transform growth factor-β_1 expression in liver tissue with confocal laser microscopy

目的探索一种能够准确定量反映肝组织转化生长因子-β1(TGF-β1)表达变化的新方法。方法以间接免疫荧光组织化学染色配合激光共聚焦显微镜扫描,对21例慢性乙型肝炎、乙肝后肝硬化患者肝组织TGF-β1的表达进行荧光定量分析,并与肝组织G-S分期半定量记分以及血清肝纤维化指标透明质酸(HA)、层黏连蛋白(LN)、Ⅲ型前胶原(PCⅢ)、Ⅳ型胶原(Ⅳ-C)等进行相关分析。结果发现肝组织TGF-β1的荧光定量分析与肝组织G-S分期半定量记分、血清肝纤维化指标等均显著相关(P<0.05),可以准确的定量反映肝组织TGF-β1的变化,反映肝纤维化的程度。结论应用间接免疫荧光配合激光共聚焦显微镜扫描是一种较为理想的定量反映肝组织TGF-β1表达及肝纤维化程度的新方法。

Objective To find a new method that can analyze transform growth factor-β1 （TGF-β1） expression quantitatively in liver tissue. Methods 21 chronic hepatitis and liver cirrhosis patients＇ serum hyaluronic acid （ HA）, laminin （ LN）, proconagen type Ⅲ （ PC Ⅲ ）, Collagen Ⅳ （Ⅳ-C） were measured by radioimmunoassay ; their liver tissues were obtained by biopsy. On the one hand, these liver slices were stained by HE, and their G-S grade was semi-quantified; on the other hand, these liver slices were used to stain the expression of TGF-β1 by indirect-immunofluorescence, analyzed by confocal laser microscopy. Result The fluorescence expression of TGF-β1 remarkably correlated with the serum liver fibrosis parameters and liver G-S grade （ P 〈 0. 05 ）. Conclusion Indirect-immunofluorescence combined with confocal laser microscopy is a new and good method to analyze TGF-β1 expression quantitatively, in liver tissue.