DADS提高HSV-tk/GCV自杀基因系统对兔晶状体上皮细胞旁观者效应的机制  

Mechanism of DADS in the bystander effect of HSV-tk/GCV suicide gene therapy system in lens epithelial cells

作　　者：李忠庆[1,2] 谭浅[1] 丁芝祥[1] 刘丹[1] 

机构地区：[1]中南大学湘雅医院眼科,长沙410008 [2]聊城市光明眼科医院,山东聊城252000

出　　处：《中南大学学报（医学版）》2011年第4期329-334,共6页Journal of Central South University ：Medical Science

基　　金：湖南省自然科学基金(06jj2071)~~

摘　　要：目的:探讨二烯丙基二硫化物(diallyl disulfide,DADS)提高重组腺相关病毒(recombinant adeno-associated virus,rAAV)介导的单纯疱疹病毒胸苷激酶(herpes simplex virus kinase,HSV-tk)/丙氧鸟苷(ganci-clovir,GCV)系统对兔晶状体上皮细胞旁观者效应的可能机制。方法:应用免疫组织化学方法检测正常兔晶状体上皮细胞间隙连接蛋白43(connexin 43,Cx43)及转染HSV-tk基因的晶状体上皮细胞Cx43的表达,以及不同浓度的DADS诱导下的2种细胞的Cx43表达,MTT比色法检测细胞存活率。结果:兔晶状体上皮细胞Cx43及转染HSV-tk基因的晶状体上皮细胞的Cx43经DADS诱导后表达阳性明显增强,尤其以50μmol/LDADS最明显。MTT比色法检测显示DADS增强了HSV-tk/GCV自杀基因系统的旁观者效应。结论:DADS可以提高晶状体上皮细胞Cx43的表达,并可提高HSV-tk/GCV自杀基因系统对兔晶状体上皮细胞的旁观者效应。A Objective To investigate the mechanism and effect of diallyl disulfide （DADS） on the bystander effect of herpes simplex virus kinase/ganeiclovir （HSV-tk/GCV） suicide gene therapy system which was mediated by recombinant adeno-associated virus （rAAV） in lens epithelial cells. Methods Immunohistochemistry method was used to determine the distribution of connexin 43 （Cx43） protein in cultured rabbit lens epithelial cells and rabbit lens epithelial cells transfected by HSV-tk suicide gene. Cx43 protein was measured and analyzed in cultured rabbit lens epithelial cells and rabbit lens epithelial cells transfected by HSV-tk suicide gene that was induced by various DADS. Cell survival was determined by 3- （4,5-dimethylthiazol-2-yl） -2,5-diphenyhetrazolium bro- mide method. Results DADS increased the Cx43 protein expression in cultured rabbit lens epithe- lial cells and rabbit lens epithelial cells transfected by HSV-tk suicide gene, and especially in 50 μmol/L DADS. After combining with DADS, the bystander effect was significantly improved （P 〈 0.05）. Conclusion DADS can elevate the expression of Cx43 protein and enhance the bystander effect of HSV-tk/GCV suicide gene therapy system.

关 键 词：二烯丙基二硫化物 旁观者效应 单纯疱疹病毒胸苷激酶 间隙连接蛋白43 

分 类 号：R346[医药卫生—基础医学]