利用单纯疱疹病毒胸苷嘧啶激酶建立肝脏特异损伤的转基因大鼠模型  

Establishment of liver injury model by specific transgenic rats with herpes simplex virus thymidine kinase

作　　者：徐涛[1] 金法[1] 李宁[1] 叶志鹏[1] 

机构地区：[1]浙江中医药大学附属第一医院普外科,杭州310005

出　　处：《中华实验外科杂志》2015年第8期2024-2027,共4页Chinese Journal of Experimental Surgery

摘　　要：目的观察单纯疱疹病毒胸苷嘧啶激酶（HSV—tk）在建立肝脏特异损伤的转丛凶大鼠模刷中的作用。方法选取鼠龄7～8周的雄性Wistar大鼠40只，通过HSV-tk载体进行转基因大鼠雄原核显微注射，获得了目的基因HSV—tk整合与特异表达的转基因大鼠，进一步通过尾静脉注射世泞洛韦（GCV）诱导转基因大鼠肝脏损伤，以四氯化碳（CCl4）损伤大鼠为对照组观察疾病的进程。通过血液的生化检查、肝脏与。肾脏等组织的病理形态学分析来研究转基因大鼠的损伤效应。结果（1）质粒pLLtk比质粒pLLtkcut对HepG2细胞有更高的杀伤效应（32％提高到53％）。（2）F1代tk转基因大鼠除肝脏与睾丸外其他组织与器官均没检测到HSV-tk的转录，说明HSV-tk的表达具有良好的组织特异性。（3）所有F1代tk大鼠在GCV处理后，经过血液生化指标与组织病理形态学检查产生了生理功能与形态学改变的肝脏损伤。（4）tk转基凶大鼠肝脏组织进行病理形态学分析，结果渺爪病理形态学普遍表现为点状或病灶性坏死，炎性细胞浸润，有凋亡细胞存在，肝细胞增生。结论成功研制肝脏表达HSV—tk的转基因大鼠，通过GCV注射诱导肝脏损伤后，产生了具有组织病州形态学与临床生物化学特征的肝细胞疾病模型。Objective To study the role of herpes simplex virus thymidine kinase （ HSV - tk） in cslablishing the transgenie rat model of liver specific injury. Methods Forty male Wistar rats of 7 - 8 weeks old were selected,the transgenic rats mieroinjection through the carrier, the expression of HSV - tk gene and the specific integration of transgenic rats ,further induce liver injury in transgenic rats by tail vein injection of GCV. Carbon tetrachloride （ CCl4 ） injury in rats as control group to observe the progress of the disease. Through biochemical tests of blood, the pathological analysis of damage of liver and kidney tissues of transgenic rats. Results （ 1 ） The plasmid pLLtk plasmid pLLtk cut than on HepG2 cell killing effect of higher（32% to 53% ）. （2）F1 generation TK transgenie rats in liver and testis and other tissues and organs were not detected the transcription of HSV - tk, indicating that HSV - tk expression is tissue specific good. 3 of all F1 tk rats ＇after GCV treatment, the blood biochemical indexes and histopathological examination of liver injury changes of physiological function and morphology. The liver tissue of 4TK transgenic rats for pathological analysis, results showed that pathological generally showed punctate or focal necrosis, in- flammatory cell infiltration, apoptosis, proliferation of liver cells. Conclusion The study has successfully developed the liver HSV -tk gene expression in rat liver cells, disease model with pathological and clinical features of biological chemical induced liver injury by GCV after injection, and provide scientific data for the study of the mechanism of liver diseases ,which laid the foundation for the study of the diagnosis and treat- merit of liver diseases.

关 键 词：肝脏特异性损伤 疱疹病毒胸苷嘧啶激酶 转基因大鼠模型 

分 类 号：R512.91[医药卫生—内科学]