单纯疱疹病毒Ⅰ型胸苷激酶基因及更昔洛韦系统对S-D大鼠的毒副作用

Toxicity studies of herpes simplex virus thymidine kinase gene(TK) and ganciclovir(GCV) system in SD rats

旨探讨将单纯疱疹病毒Ⅰ型胸苷激酶基因载体生产细胞（pLTKcSN／VPC）脑内注射和腹腔注射更昔洛韦（GCV）在大鼠体内的急性系统性毒副作用，以及载体生产细胞（VPC）在大鼠脑内的存在时问。S-D大鼠24只，雌雄各半，随机分为2组。第1组颅内注射pLTKcSN／VPC，细胞总量3ｘ106/只，7天后腹腔注射GCV30mg·kg-1·d-1，共7天。GCV用药结束后1周处死动物，进行备脏器组织学检查及TK序列的聚合酶链反应（PCR）和原位杂交检测。第2组，采用脑内注射等量整合有β半乳糖苷酶基因（β-gal／nVPC），注射后的9、14、21和30天各处死3只动物，取注射针道周围脑组织行冰冻切片x-gal组化染色，观察VPC在大鼠脑内的生存时限。结果：第1组动物的各脏器组织学病理改变主要为脑内手术部位蛛网膜下腔出血、个别动物有心肌间质灶性炎症和肝细胞脂肪变性。TK序列聚合酶链反应（PCR）检测动物体内各脏器均未检出TK序列。x-gal组化染色见9天和21天两个时问点分别有1只动物脑内有阳性细胞存在。本文结果提示：pLTKcSN／VPC及GCV系统对大鼠脑及全身各脏器有轻微非特异性急性毒副作用，pLTKcSN／VPC在大鼠体内的生存时间约为3周。为pLTKCSN／VPC及GCV系统的临床试验治疗恶性脑胶质瘤提供了脑内注射PLTK。SN／VPC的安全性依据。

To evaluate the toxicity effects of retroviral-mediated herpes simplex virus thymidine kinase gene (TK) transfer and ganciclovir in rats as well as the life span of retrovirus producer cells in situ. Twenty-four SD rats wererandomly divided into two groups. Rats in group 1 received TK vector producer cells(pLTKcSN /VPC ) 3 ｘ l06 throughstereotactic intracerebral injection,rats in group 2 were injected with Lacz gene marked PA3l7 cells (β-gal/nVPC) with the same dose as that of group l. Seven days later, animals in group l were given ganciclovir 30mg. kg-1. d-1 intra-peritoneally for another 7 days. The animals were then sacrificed, historical examination as well as PCR and hybridiza-tion in situ detection of TK sequence of all organs were carried out. The animals in group 2 were sacrificed on the 9, 12,2l and 30 days after β-gal/nVPC injection (3 rats at each timepoint, x-gal histochemical stairing of the brain slices wereperformed. In group l, no significant pathological changes could be detected in all organs, PCR examination of TK se-quence also showed negative results. The in situ hybridization of TK sequence revealed that there was sporadic TK posi-tive cells inthe heart, lung, liver and colon mucous membrane. In group 2, the x-gal examination unveiled that there were x-gal positive cells on day 9 and 21 after cells injection, but the staining results of the 30th days were negative. The pLTKcSN/VPCand GCV system has slight toxicity effects on brain, heart, liver and lung,the life span of the VPC in rat brain is about 3 weeks.