腺病毒载体在小鼠肝脏的分布及其表达规律研究

Distribution and expression of adenovirus vector in mice with MHV-3-induced fulminant hepatitis

目的研究腺病毒载体介导的β-半乳糖苷酶(LacZ)报告基因在小鼠肝脏的分布及表达情况,评价腺病毒载体作为基因治疗转导系统的有效性及安全性,为进一步的目的基因导入及表达奠定基础。方法将Balb/cJ小鼠随机分为正常组及暴发性肝炎组,给小鼠经尾静脉注射2×108PFU腺病毒,收集24h、48h、72h、96h、120h和第7天小鼠血清,检测谷丙转氨酶和总胆红素,同时取肝脏、肺脏、心脏、肾脏标本进行X-gal染色,以观察腺病毒载体在各脏器的表达情况。取MHV-3诱导的暴发性肝炎组小鼠24h、48h、72h、96h肝脏,检测腺病毒载体在肝脏的表达情况。结果腺病毒载体主要在小鼠肝脏表达,并于72小时表达最高,在正常小鼠及暴发性肝炎小鼠肝脏中表达效率分别约55.7%和25.7%,之后逐渐消减;正常组小鼠注射腺病毒载体后,肝功能无明显变化。结论腺病毒载体介导的报告基因可在肝脏高效表达且未见明显的肝损害,可作为基因治疗中安全有效的基因传递系统用于治疗肝脏疾病。

Objective To study the distribution and expression of β-galactosidase （LacZ） reporter gene mediated by adenovirus vector in the liver of normal mice or mice with fulminant hepatitis, so as to evaluate the efficacy and safety of adenovirus as a gene therapy vector transduction system. Methods The normal Balb/cJ mice were randomly assigned to normal and fulminant hepatitis group. Each mouse was injected with adenovirus at 2×10^8PFU by tail vein injection,then the normal mice serum was collected to detect ALT and TBil at 24h,48h, 72h,96h,120h, and the seventh day,meanwhile,the mice organs including liver,lung,heart,and kidney were collected and stained by X-gal in order to observe the expression of adenovirus vector in mice organ;the expression of adenovirus vector in liver of fulminant hepatitis mice induced by MHV-3 was detected. Results Adenovirus mainly expressed in the livers of mice;In normal group and fulminant hepatitis group,the highest expression efficiency was at 72 hours,reached 55.7% and 25.7%,respectively,then it followed by a gradual reduction of adenovirus expression; The adenovirus infection didn＇t show obvious injury in liver function. Conclusions The adenovirus vector is highly expressed in the liver with no significantly liver damage; Adenovirus vector might be as an effective and safe gene delivery system for liver disease gene therapy.