胆道闭锁肝内外胆系免疫炎性反应与临床预后相关分析

目的 测定胆道闭锁(BA)肝组织和肝门纤维块中白细胞分化抗原(CD68)、单核细胞趋化蛋白-1(MCP-1)和细胞间粘附因子-1(ICAM-1)的含量,分析肝内外组织免疫炎性改变与临床预后相关性,旨在了解其在发病过程的重要性.方法 取2005年7月至2006年5月在本院行BA Kasai根治术的21例患儿的肝门纤维块及肝脏组织标本;对照组为5例胆汁淤积综合征患儿和10例胆总管囊肿患儿的肝脏组织标本.采用免疫组化染色法进行CD68、MCP-1和ICAM-1染色.选用LEICA DM研究级生物显微镜,QWIN软件环境下测量抗体阳性细胞面积百分比和平均光密度.随访19例BA患儿术后3个月直接胆红素下降比率,与肝组织CD68、MCP-1和ICAM-1表达量进行相关性分析.结果 BA组肝组织CD68、MCP-1、ICAM-1表达较对照组明显增强,但肝门纤维块几乎没有表达;CD68、MCP-1、ICAM-1阳性表达量和表达强度明显高于对照组,其中ICAM-1最为显著;除了MCP-1外,CD68、ICAM-1在肝脏表达的表达量与表达强度呈线性正相关,相关系数分别是r=0.838,P=0.0002;r=0.565,P=0.035;BA组肝组织CD68阳性表达面积百分比与术后3个月直接胆红素下降率呈负相关(r=-0.645,P=0.023).结论 CD68阳性表达是BA免疫炎性反应过程的一个重要环节,MCP-1和ICAM-1在这一环节中起了一定的作用;肝组织CD68阳性表达可能提示预后不佳.

The correlative study of the intra-and extrahepatic biliary system inflammation and prognosis

Objective To detect the levels of CD68,MCP-1,and ICAM in the liver and porta hepatis fibrous tissue biliary atresia(BA),and investigate the relationship between intra-and extrahepatic biliary system inflammation and prognosis.Methods From Sept 2005 to May 2006,21 cases with BA underwent Kasai procedure.Biopsy of the liver and triangular cord of porta hepatis was performed in operation.As the control group,intraoperative biopsy was also performed in 5 cases with cholestasis syndrome and 10 cases with choledochal cyst. The levels of CD68,MCP-1,and ICAM in BN's liver and porta hepatis fibrous tissue were assessed via two-step immunohistochemistry.Leica DM microscope was used to obtain the immunohistochemistry image.Positive cells were counted in a complete section,and the total area of the section and median integrated optical density(IOD)of the positive cells were measured by Qwin Image Analysis software(Leica Image Systems).Expression level was defined as positive area proportion,and expression density was IOD.Direct bilirubin of the 19 cases with BA was followed up.The relationship between declining rate of direct bilirubin and the amounts of CD68,MCP1,and ICAM was assessed.SPSS software was used for statistical analysis.Results (1)CD68 was expressed in cytoplasm of liver Kupffer's cells,but not in the porta hepatis fibrous tissues.ICAM was expressed in bile duct epithelial cells and hepatocytes,but MCP-1 was rarely seen in porta hepatic.(2)All of the 3 antigens were expressed higher in BA group than those in control group,and ICAM was expressed much higher than the other two antigens.Levels of CD68 and ICAM were positively linear correlated with the intensities of them.Correlation coefficient was r=0.838,P=0.0002(CD68)and r=0.565,P=0.035(ICAM).(3)The levels of CD68 in the liver of BA were negatively correlated with the declining rate of direct bilirubin 3 months postoperatively.Correlation coefficient was r=-0.645,P=0.023.Conclusions CD68 plays an important role in the process of intra-and extrahepatic biliary system inflammation,and MCP and ICAM also participate in that process.High CD68 expression suggests bad prognosis of patients with BA.