肿瘤坏死因子-α和半胱氨酸天冬氨酸蛋白酶3在肝泡型棘球蚴周围单核细胞中的表达

中国寄生虫学与寄生虫病杂志 ›› 2012, Vol. 30 ›› Issue (3): 7-201-205.

肿瘤坏死因子-α和半胱氨酸天冬氨酸蛋白酶3在肝泡型棘球蚴周围单核细胞中的表达

杨宏强1,2，马少波1，卞志远1，李江1，邹泓3，张示杰1,2，彭心宇1,2 ，陈孝平2 *

1石河子大学医学院第一附属医院肝胆外科，石河子 832002； 2华中科技大学同济医学院附属同济医院肝脏外科中心，武汉430030；
3石河子大学医学院病理学教研室，石河子 832002

出版日期:2012-06-30 发布日期:2012-09-28

Expression of Tumor Necrosis Factor-α and Caspase-3 Protein in Monocytes Adjacent to the Invaded Echinococcus multilocularis in Liver

YANG Hong-qiang1,2, MA Shao-bo1, BIAN Zhi-yuan1, Li Jiang1, ZOU Hong3, ZHANG Shi-jie1,2, PENG Xin-yu1,2, CHEN Xiao-ping2*

1 Department of Hepatobiliary Surgery, the First Affiliated Hospital, Medical College of Shihezi University, Shihezi 832002, China; 2 Hepatic Surgery Center, Tongji Hospital, Huazhong University of Science and Technology, Wuhan 430030, China; 3 Department of Pathology, Medical College of Shihezi University, Shihezi 832008, China

Online:2012-06-30 Published:2012-09-28

摘要/Abstract

摘要： 目的探讨肿瘤坏死因子-α（TNF-α）和半胱氨酸天冬氨酸蛋白酶3 (caspase-3) 在肝泡型棘球蚴周围单核细胞中的表达。方法 40只雌性昆明小鼠随机分为实验组 (n=20) 和假手术对照组 (n=20)，实验组小鼠开腹直视下肝脏穿刺注射0.1 ml泡型棘球蚴混悬液，对照组注射等量生理盐水。感染6个月后处死小鼠，取肝组织，观察泡型棘球蚴的生长和转移情况；用苏木素-伊红 (HE) 染色法观察组织病理变化；免疫组织化学法检测TNF-α和caspase-3在肝泡型棘球蚴和淋巴转移灶周围组织中的表达；原位末端标记技术 (TUNEL) 检测肝组织和泡型棘球蚴病灶周围单核细胞的凋亡。结果感染6个月后，实验组小鼠肝脏可见大小不等的结节状或团块状的泡型棘球蚴组织，淋巴结转移率为45.0% (9/20）。HE染色后观察发现，实验组肝泡型棘球蚴和淋巴转移灶周围均有大量单核细胞浸润。免疫组化染色结果显示，实验组TNF-α和caspase-3蛋白在肝泡型棘球蚴和淋巴转移灶周围均有不同程度的阳性表达，单核细胞阳性细胞检出率分别为100%、100%和95%、100%，与对照组肝组织中阳性细胞检出率（5%和0）比较均有统计学意义（P＜0.01）。TUNEL结果显示，肝泡型棘球蚴和淋巴转移灶周围单核细胞发生凋亡，单核细胞阳性细胞检出率为100%，与对照组比较有统计学意义（P＜0.01）。结论泡型棘球蚴在感染宿主过程中，引起TNF-α蛋白高表达，可能参与了诱导肝泡型棘球蚴周围宿主单核细胞的凋亡，从而抑制宿主的免疫功能。

关键词: 泡型棘球蚴病；单核细胞；凋亡；肿瘤坏死因子-&alpha

Abstract: Objective To investigate the expression and its significance of tumor necrosis factor-α (TNF-α) and caspase-3 protein in monocytes adjacent to the invaded Echinococcus multilocularis in liver. Methods 40 female Kunming mice were randomly divided into experimental group (n=20) and sham operation (control) group (n=20). Mice in experimental group were infected with 20% E. multilocularis suspension (0.1 ml per mouse) through abdominal opening injection in liver and the mice in control group were injected with equal physiological saline. The mice were sacrificed at 6 months post-infection for observing the growth and metastasis of E. multilocularis. Pathological changes were observed by HE staining． The expression of TNF-α and caspase-3 protein in hydatid cyst and metastasis tissue were detected by immunohistochemistry staining and the apoptosis of the monocytes was measured by TUNEL． Results After 6 months post-infection, E. multilocularis were spread over the liver of the mice in experimental group. Metastasis rate of lymph nodes was 45.0%（9/20）. Infiltration of monocytes was observed around E. multilocularis in liver and lymph nodes with metastasis by HE staining. Immunohistochemistry showed that the positive expression rate of TNF-α and caspase-3 protein in monocytes was 100% and 100%, and 95% and 100% respectively around the cyst in experimental group, while the expression rate was only 5% and 0 respectively in the liver of the control mice (P＜0.01). The monocytes showed significant apoptosis by TUNEL in experimental group with a positive expression rate of 100%, with a significant difference between experiment group and the control (P＜0.01). Conclusion In the process of alveolar Echinococcus infection, the high expression of TNF-α protein might be associated with the apoptosis of monocytes, which may inhibit the host immunological function.

Key words: Alveolar echinococcosis； Monocyte； Apoptosis； Tumor necrosis factor-&alpha

杨宏强1,2，马少波1，卞志远1，李江1，邹泓3，张示杰1,2，彭心宇1,2 ，陈孝平2 *. 肿瘤坏死因子-α和半胱氨酸天冬氨酸蛋白酶3在肝泡型棘球蚴周围单核细胞中的表达[J]. 中国寄生虫学与寄生虫病杂志, 2012, 30(3): 7-201-205.

YANG Hong-qiang1,2, MA Shao-bo1, BIAN Zhi-yuan1, Li Jiang1, ZOU Hong3, ZHANG Shi-jie1,2, PENG Xin-yu1,2, CHEN Xiao-ping2*. Expression of Tumor Necrosis Factor-α and Caspase-3 Protein in Monocytes Adjacent to the Invaded Echinococcus multilocularis in Liver[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2012, 30(3): 7-201-205.