CAJ | 학술논문

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基因大鼠模型中的作用.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更昔洛韦(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pLLtk cut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转基因大鼠肝脏组织进行病理形态学分析,结果显示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,产生了具有组织病理形态学与临床生物化学特征的肝细胞疾病模型.
목적 관찰단순포진병독흉감밀정격매(HSV-tk)재건립간장특이손상적전기인대서모형중적작용.방법 선취서령7～8주적웅성Wistar대서40지,통과HSV-tk재체진행전기인대서웅원핵현미주사,획득료목적기인HSV-tk정합여특이표체적전기인대서,진일보통과미정맥주사경석락위(GCV)유도전기인대서간장손상,이사록화탄(CCl4)손상대서위대조조관찰질병적진정.통과혈액적생화검사、간장여신장등조직적병리형태학분석래연구전기인대서적손상효응.결과 (1)질립pLLtk비질립pLLtk cut대HepG2세포유경고적살상효응(32％제고도53％).(2)F1대tk전기인대서제간장여고환외기타조직여기관균몰검측도HSV-tk적전록,설명HSV-tk적표체구유량호적조직특이성.(3)소유F1대tk대서재GCV처리후,경과혈액생화지표여조직병리형태학검사산생료생리공능여형태학개변적간장손상.(4)tk전기인대서간장조직진행병리형태학분석,결과현시병리형태학보편표현위점상혹병조성배사,염성세포침윤,유조망세포존재,간세포증생.결론 성공연제간장표체HSV-tk적전기인대서,통과GCV주사유도간장손상후,산생료구유조직병리형태학여림상생물화학특정적간세포질병모형.
Objective To study the role of herpes simplex virus thymidine kinase (HSV-tk) in establishing the transgenic rat model of liver specific injury.Methods Forty male Wistar rats of 7-8 weeks old were selected,the transgenic rats microinjection 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 transgenic 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,inflammatory 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 treatment of liver diseases.