华支睾吸虫感染致胆管损伤肝纤维化的实验研究

摘要/Abstract

摘要： 目的探讨建立家兔华支睾吸虫感染模型时,不同感染剂量对家兔胆管和肝细胞损伤的差异,为造模提供参考依据。方法采用人工消化法提取华支睾吸虫囊蚴,分别以400、600、800个囊蚴经口灌胃15只家兔,设为低、中、高感染组,同时设生理盐水对照组(5只家兔）,感染90天后取动物肝脏标本行病理学检查。结果经囊蚴灌胃的家兔均成功感染华支睾吸虫,感染后4周粪便虫卵检出率达100%,低、中、高感染组成虫回收率最高分别为20.50%、15.17%和23.38%,低、中感染组间成虫数量比较差异无统计学意义（P=0.24）,低、中感染组与高感染组间成虫数量比较差异有统计学意义（P<0.01）。感染组家兔肝内胆管明显增粗,在肝内胆管和胆囊中找到大量活成虫;镜下见汇管区增大并有胶原成分积聚,肝内胆管增粗、胆管壁增厚,胆管及周围组织炎症,胆管内皮局部可见低级到高级的上皮内瘤变;Masoon染色观察,按Metavir肝纤维化评分,对照组5只均为肝纤维化F0期,实验组2只为F1期,4只为F2期,4只为F3期,低感染组与高感染组之间肝纤维化分期差异有统计学意义（P<0.05)。结论造模90天,600个囊蚴为家兔华支睾吸虫感染的最佳推荐剂量,满足胆管损伤、肝纤维化的实验造模要求。

关键词: 华支睾吸虫, 模型, 兔, 胆管损伤, 肝纤维化

Abstract: Objective To explore the difference of bile duct and hepatocyte damage caused by different doses of Clonorchis sinensis infection in rabbits, and to provide reference for the establishment of models.Methods The metacercarias of clonorchis sinensis were extracted by artificial digestion, collected and counted under microscope. 400, 600 and 800 metacercarias were given to 15 rabbits by oral in 3 groups named low-degree, medium-degree and high-degree infected group, normal saline was given to 5 rabbits which were served as control group. All rabbits were sacrificed after 90 days and the liver specimens were performed with pathological examination.Result sRabbits in 3 infected groups were all infected successfully. Egg positive rates in feces reached 100% in 4 weeks post infection. The highest adult Clonorchis sinensis recovery rate in low, medium and high infected-groups were 20.50%,15.17% and 23.38%. There was no statistical difference in the number of adults between the low and medium infection groups（P=0.24）,the comparison of number of adults between low and medium infected-groups with high infected-group showed significant differences（P<0.01）. The rabbit liver in the infected group was found that biliary ducts in the liver were thickened significantly, and a large number of live adult Clonorchis sinensis were found in the intrahepatic bile ducts and gallbladder; Observed with light microscope, the portal area was enlarged with collagen accumulation, the intrahepatic bile duct was thickened, the bile duct wall was thickened, the bile duct and surrounding tissues were inflamed, and low to high level intraepithelial neoplasia was observed in the endothelium of the bile duct. Masoon trichromic staining was observed and stage of fibrosis was quantified according to the Metavir scoring system -- liver fibrosis staging score. In the control group, 5 rabbits were all in the stage of F0 (no liver fibrosis), while in the infected group 2 were in the stage F1, 4 were in the stage F2, and 4 were in the stage F3, There was statistical difference in the stage of fibrosis between low and high infection groups（P<0.05）.Conclusions Infection for 90 days, the best recommended dose for Clonorchis sinensis infection model in rabbit is 600 metacercarias, it was suitable to the requirements for experimental model as biliary duct injury and hepatic fibrosis.

Key words: Clonorchis sinensis, Animal model, Rabbit, Biliary duct injury, Hepatic fibrosis

方竞, 周忠洋, 廖锦元. 华支睾吸虫感染致胆管损伤肝纤维化的实验研究[J]. 新发传染病电子杂志, 2019, 4(2): 112-116.

FANG Jing, ZHOU Zhong-yang, LIAO Jin-yuan. Experimental study on Clonorchis sinensis infection induced biliary duct injury and hepatic fibrosis[J]. Electronic Journal of Emerging Infectious Diseases, 2019, 4(2): 112-116.

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