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  

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

作　　者：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] 

机构地区：[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　　处：《中国寄生虫学与寄生虫病杂志》2012年第3期201-205,共5页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金资助项目(No.81160199);国家科技部支撑计划项目(No.2009BAI82B06);新疆生产建设兵团医药卫生科技攻关项目(No.2010GG54);石河子大学重大科技攻关项目(No.gxji2008-zdgg06)~~

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cas-pase-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。Objective To investigate the expression and its significance of tumor necrosis factor-or （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 metastasisby HE staining. Immunohistochemistry showed that the positive expression rate of TNF-α and caspase-3 protein in mono- cytes 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.

关 键 词：泡型棘球蚴病 单核细胞 凋亡 肿瘤坏死因子-Α 

分 类 号：R383-33[医药卫生—医学寄生虫学]