慢性乙型肝炎血瘀病机与肝脏纤维组织沉积及血管新生病理变化关系研究

Investigation of pathological changes of hepatic fibrous tissue deposition and angiogenesis in chronic hepatitis B with blood stasis

目的探讨肝组织胶原纤维沉积、CD34及细胞角蛋白19(cytokeratin 19,CK19)水平对慢性乙型肝炎血瘀病机判识的价值,以期为中医辨治慢性乙型肝炎提供参考。方法以128例行肝组织活检术的慢性乙型肝炎患者为研究对象,根据中医四诊信息将患者分为血瘀证组和非血瘀证组。对患者肝组织切片进行苏木素-伊红、网状纤维、Masson染色,以及CD34及CK19免疫组化染色;采用图像分析软件计算肝组织切片全片胶原纤维(Masson染色)、CD34及CK19的阳性表达水平,比较血瘀证组和非血瘀证组肝组织胶原纤维、CD34及CK19表达差异。结果①肝组织纤维化分期S2、S3和S4期的胶原纤维、CD34和CK19阳性面积显著大于S1期(P<0.05),S4期的胶原纤维和CD34的阳性面积显著高于S2和S3期(P<0.05)。②炎症活动度G4级的胶原纤维阳性面积显著大于G1级、G2级和G3级(P<0.05),G1级、G2级和G3级之间差异无统计学意义(P>0.05);不同炎症活动度分级之间的CD34和CK19阳性表达差异无统计学意义(P>0.05)。③血瘀证与非血瘀证患者的纤维化分期差异有统计学意义(P<0.05),血瘀证患者胶原纤维、CD34阳性表达显著高于非血瘀证患者(P<0.05)。结论慢性乙型肝炎患者肝组织胶原纤维与CD34阳性表达可作为肝纤维化程度的量化指标;胶原纤维沉积及血管新生可能是慢性乙型肝炎血瘀病机肝病理组织学的主要变化,可为中医辨证治疗提供部分参考依据。

Objective To explore the value of collagen fibers deposition,CD34 and Cytokeratin 19( CK19) expression in liver tissue in the diagnosis of chronic hepatitis B with blood stasis syndrome,in order to provide some reference for traditional Chinese medicine( TCM) syndrome differentiation and treatment on chronic hepatitis B.Methods Research objects were 128 hepatitis B patients undergoing liver biopsy. According to TCM four diagnostic information,the patients were divided into blood stasis syndrome group and non-blood stasis syndrome group.Hematoxylin-eosin staining,reticular fiber staining,Masson staining,CD34 and CK19 immunohistochemical staining were performed on the liver tissue sections,and then the positive expressions of collagen,CD34 and CK19 were calculated by the image analysis software to observe and compare the changes of collagen fiber,CD34 and CK19 expression in liver tissue. Results(1) The positive expressions of collagen fibers, CD34, and CK19 in patients with liver fibrosis stages S2,S3 and S4 were significantly higher than those in patients with S1 stage( P<0.05). The expression of collagen fibers and CD34 in patients with S4 stage were significantly higher than those in stages S2 and S3( P<0.05).(2)The positive area of collagen fibers in G4 degree of inflammatory activity was significantly larger than those in G1,G2 and G3 degree( P<0.05). There was no significant difference between G1,G2 and G3 degree( P>0.05) and no significant difference in positive expression of CD34 and CK19 between different degrees of inflammatory activity( P>0.05).(3) There was a significant difference in fibrosis stages between patients with blood stasis syndrome and non-bloody stasis syndrome( P<0.05). The positive expression of collagen fiber and CD34 in patients with blood stasis syndrome was significantly higher than that in patients without blood stasis syndrome( P<0.05). Conclusion Positive expression of collagen fiber and CD34 in liver tissue of patients with chronic hepatitis B can be a quantitative index of liver fibrosis. Collagen fiber deposition and angiogenesis may be the main changes in liver histopathology of chronic hepatitis B with blood stasis syndrome,which may provide some references for TCM syndrome differentiation and treatment on chronic hepatitis B.