阿司匹林中断硫代乙酰胺诱发大鼠胆管上皮癌变

Aspirin interrupts bile duct carcinoma in rats induced by thioacetamide

目的构建胆管癌原位诱导大鼠模型,应用阿司匹林干预该模型,观察阿司匹林对胆管癌的影响。方法给予SD雄性大鼠饮用含有硫代乙酰胺(300 mg/L)的饮用水,第8、12、16和20周时H-E染色分别观察肝脏成瘤情况,采用免疫组化法检测碳酸酐酶2(CA-2)在肝脏组织中的表达。另选取SD雄性大鼠,在硫代乙酰胺饲养12周后开始用阿司匹林处理,第3、6个月取出肝脏,H-E染色观察肝脏成瘤情况,免疫组化法检测CA-2的表达变化。取胆管癌细胞QBC939,予以阿司匹林(0 mmol/L和5 mmol/L)处理,培养48 h后采用蛋白质印迹法检测细胞中CA-2的表达。结果硫代乙酰胺饲养第12周时部分大鼠肝内可见明显的纤维化;第16周时所有大鼠肝内均出现大量纤维组织增生,镜下可见肿瘤或疑似肿瘤;第20周时所有大鼠肝脏均形成肉眼和镜下可见的肿瘤。阿司匹林处理6个月后,大鼠肝内肿瘤或疑似肿瘤的发生率为28.6%(2/7),而未处理组中大鼠肝内肿瘤或疑似肿瘤的发生率为100%(7/7)。大鼠肝脏组织中CA-2的表达水平随胆管癌病情的进展逐渐升高,而阿司匹林处理后大鼠肝脏组织中CA-2的表达减弱。5 mmol/L阿司匹林处理48 h后QBC939细胞中CA-2的表达与未处理对照组相比下调(P<0.01)。结论硫代乙酰胺能成功诱导肝内胆管癌的发生,阿司匹林能阻断该过程,可用于预防肝内胆管癌。

Objective To establish a thioacetamide(TAA)-induced bile duct carcinoma rat model and to observe the effect of aspirin on bile duct carcinoma by interrupting the model.Methods Sprague-Dawley(SD)male rats were given drinking water containing TAA(300 mg/L)for 5 months,and tumor formations of the livers were observed by H-E staining at the 8th,12th,16th,and 20th weeks,respectively.The expression of carbonic anhydrase 2(CA-2)in liver tissues was also detected by immunohistochemistry.SD male rats were also selected and treated with aspirin after exposing to TAA for 12 weeks.And then the tumor formations of the livers were observed by H-E staining at the 3rd and 6th months,and the expression of CA-2 was detected by immunohistochemistry.Bile duct carcinoma cells QBC939 were obtained and treated with aspirin(0 mmol/L and 5 mmol/L),and the expression of CA-2 was detected by Western blotting after culturing for 48 h.Results Obvious fibrosis was found in the livers of some rats at 12th weeks after exposing to TAA;large number of fibrous tissue hyperplasia and microscopic or suspected tumors were found in the livers at the 16th weeks;visible tumors in the livers were found in all the rats at the 20th weeks.The incidence of hepatic tumors or suspected tumors was 28.6%(2/7)in rats treated with aspirin for 6 months,while it was 100%(7/7)in rats without aspirin treatment.The expression of CA-2 in liver tissues was gradually increased with the development of bile duct carcinoma in rats,while the expression of CA-2 in liver tissues was decreased after treating with aspirin.The expression of CA-2 in QBC939 cells treated with 5 mmol/L aspirin for 48 h was significantly decreased versus the untreated control group(P<0.01).Conclusion TAA can successfully induce intrahepatic bile duct carcinoma.Aspirin can interrupt the development of bile duct carcinoma and may be used to prevent intrahepatic bile duct carcinoma.