胆道闭锁肝组织CD14、CD34、TGF-β1表达研究

Expression of CD14, CD34 and transforming growth factor-β1 in liver biopsy of biliary atresia

目的探讨肝组织细胞表面糖蛋白14（CD14），血管内皮生长因子34（CD34），转化生长因子β1（TGF-β1）在胆道闭锁（Biliary atresia，BA）肝组织中的表达及其在肝纤维化中的作用。方法选取2013年1月至2014年1月我院BA患）L（Kasai组）、外院BA接受肝移植自体肝组织（移植组）、胆管扩张症（扩张组，作为对照组），每组15例，术中留取肝组织。进行HE及免疫组化染色检测三种蛋白在肝组织中表达及组织定位情况，分析其与肝纤维化之间的关系。应用图像分析软件（Image-Pro-Plus）分别计算CD14，TGF-β1积分光密度值（IOD），CD34阳性表达面积（Area），SPSS17．0软件统计分析三组蛋白之间表达差异及其相关性。结果CD14表达于肝髓窦内皮细胞，成纤维细胞，炎细胞；TGF-β1表达于成纤维细胞、胆管上皮细胞、炎细胞；CD34表达于汇管区血管内皮细胞。与对照组比较，实验组三种蛋白表达增加，从肝纤维化Ⅰ级至Ⅳ级中，CD14、TGF-β1蛋白表达量呈逐渐下降趋势，CD34表达逐步增多。实验组中三种蛋白在肝组织表达差异有统计学意义（P〈0．05），其中CD14与TGF-β1肝组织表达相关性显著，TGF-β1、CD34肝组织表达呈正相关（P〈0．01）。结论BA肝纤维化进展期CD14，TGF-β1表达逐渐增加，参与BA肝纤维化过程，并对BA肝硬化形成起到推动作用。到移植终末期，各种细胞缺失，纤维组织增多，三组蛋白表达逐渐下降。

Objective To explore the expression of surface glyeoprotein protein hepatocyte 14 （CD14）, vascular endothelial growth factor 34 （CD34） and transforming growth factor-β1 （TGF-β1） in liver biopsy of biliary atresia （BA） and examine the role of liver fibrosis. Methods From January 2013 to January 2014, liver biopsy specimens were collected intraoperatively from BA patients undergoing Kasai procedure （Kasai group）, BA patients after Kasai procedure with liver transplantation （transplantation group） and congenital biliary dilation patients （CBD group as control） （n = 15 each）. The hematoxylin ＆ eosin staining and immunohistoehemistry were used to observe the degree of liver fibrosis and detect the expression of CD14, CD34 and TGF-β1 and determine their localizations. The image analysis software of Image-Pro-Plus was used to analyze the values of integrated optical density （IOD） for CD14 and TGF-β1 proteins in different groups and CD34 positive area. SPSS 17. 0 statistical software was used for single factor ANOVA analysis （P〈0. 05）. Pearson correlation analysis （P〈0. 01） showed that there was correlation between three groups about the expression of different proteins. Results CD14 was expressed in myeloid liver sinusoidal endothelial cells, fibroblasts and inflammatory cells. And TGF-β1 was expressed in fibroblasts, biliary epithelial cells and inflammatory cells while CD34 in portal area and proliferating vascular endothelial cells. Compared with CBD group, the expressions of CD34, CD14 and TGF-β1 proteins were up-regulated in experimental group. From the perspective of liver fibrosis grading （stage I-IV） in BA. The expression of CD14 and TGF-β1 proteins decreased gradually while the expression of CD34 increased. The difference was statistically significant between three groups about the expression of CD14, TGF-β1 and CD34 proteins （P〈0. 05）, the expression of CD14 and TGF-β1 proteins were significantly correlated （P〈0. 05） whereas the expression of TGF-β1 and CD34 proteins significantly positively correlated.Conclusions The expression of CD14 and TGF-β1 in liver fibrosis increase gradually in B.A. And liver cirrhosis may be accelerated. In advanced disease, all cells are damaged and become substituted by fibrosis. And the levels of three proteins gradually decline.