二乙基亚硝胺诱导大鼠原位肝癌模型及在TACE中应用研究

Diethylnitrosamine-induced rat in situ liver cancer model and its application in transarterial chemoembolization therapy

目的 评价二乙基亚硝胺(DEN)口服诱导的大鼠肝癌模型的病理学特性及其在经导管动脉化疗栓塞术(TACE)中的应用价值。方法 60只Sprague-Dawley(SD)大鼠口服0.01%DEN诱导原位肝癌,分为肿瘤模型组(n=40)和TACE应用组(n=20)。肿瘤模型组大鼠自诱导后第6周开始每周行肝脏MRI检查,肝脏出现0.2 cm、0.5 cm结节时分别随机处死5只大鼠,行肝脏大体观、苏木精-伊红(HE)及Masson染色检查;其余大鼠继续每周行肝脏MR及每2周肺部CT检查,观察肿瘤生长、转移,并记录大鼠生存时间。TACE应用组大鼠采用开腹经胃十二指肠动脉逆行插管行介入操作,记录技术成功率和死亡率;术后3 d、7 d分别处死大鼠,检测肿瘤坏死。结果 肿瘤模型组大鼠肿瘤结节直径达0.2 cm的中位诱导时间为8(7,10)周,肝脏体积增大,肿瘤呈灰白色微小结节;HE染色显示肝实质炎性细胞浸润,部分假小叶形成,肿瘤细胞核肥大深染;Masson染色显示少量胶原纤维。肿瘤结节直径达0.5 cm的中位诱导时间为11(10,12)周,肝脏体积缩小,表面有大量结节;HE染色可见假小叶形成;Masson染色显示胶原纤维增多。部分大鼠出现肝外转移。中位生存时间为诱导后13(8,20)周。TACE应用组大鼠经DSA证实肿瘤为富血供;TACE技术成功为11/16,手术相关死亡为3/16;术后3 d、7 d肿瘤坏死率分别为(48.16±2.65)%、(50.36±2.92)%。结论 DEN诱导的大鼠原位肝癌较好地模仿了人类肝癌组织病理学特性,可用于TACE操作,是一种理想的介入治疗动物模型。

Objective To establish a diethylnitrosamine-induced(DEN-induced) rat in situ liver cancer model, and to evaluate its pathological characteristics and its application value in transarterial chemoembolization(TACE) therapy. Methods A total of 60 Sprague-Dawley(SD) rats were used in this study. The DEN of 0.01%concentration was orally administered to all the experimental rats to induce in situ liver cancer. The experimental rats were divided into tumor model group(n=40) and TACE treatment group(n=20). For the rats of tumor model group, liver MR imaging was performed once a week, which started from the 6th week after DEN administration, and every 5 randomly selected rats were sacrificed each time when the diameter of liver nodule reached 0.2 cm and 0.5 cm large. The liver samples were macroscopically examined, and after HE and Masson staining microscopical examination was performed. The remaining rats continued to receive liver MRI weekly and lung CT examination ones every 2 weeks to check the status of tumor growth and metastasis, and the survivals of rats were recorded. For the rats of TACE treatment group, laparotomy chemoembolization through retrograde catheterization via gastroduodenal artery was performed. The technical success rate,procedure-related mortality were documented. At 3 and 7 days after treatment, the rats were sacrificed and the tumor necrosis was evaluated. Results In tumor model group, the median induction time of liver nodule diameter reaching 0.2 cm was 8 weeks(7, 10 weeks). The macroscopic examination showed that the liver volume was enlarged, the tumor lesions presented as grey-white tiny nodules. After HE staining, in the liver parenchyma the inflammatory cell infiltration, formation of false lobules, and larger, darker-stained tumor nucleus were observed. After Masson staining, small amount of collagen fibers was demonstrated. The median induction time of liver nodule diameter reaching 0.5 cm was 11 weeks(10, 12 weeks). Macroscopically, the liver volume was reduced with numerous nodules on its surface. HE staining demonstrated the formation of false lobules, and Masson staining revealed that the collagen fibres were increased. Extrahepatic metastases were detected in some rats. The median survival time was 13 weeks after induction(8, 20 weeks). In TACE treatment group, DSA confirmed that the tumors were hypervascular lesions. The technical success rate of TACE was68.8%(11/16), the operation-related mortality was 8.8%(3/16). The postoperative 3-day and 7-day necrosis rates were(48.16±2.65)% and(50.36±2.92)% respectively. Conclusion DEN-induced rat in situ liver cancer model better mimics the histopathological characteristics of human liver cancer, which can be used in TACE operation, and this model is an ideal animal model for interventional research.(J Intervent Radiol, 2022,31: 686-692)