腺病毒对原代枯否细胞转染效率及增殖活性的影响  

Impacts of adenovirus on the transfection efficiency and proliferative activity of primary Kupffer cells

作　　者：朱彤[1] 涂文娟[1] 谈志丽 刘亮明[1] 

机构地区：[1]南京医科大学附属上海松江中心医院感染科,上海市201600

出　　处：《实用医学杂志》2016年第6期879-882,共4页The Journal of Practical Medicine

基　　金：国家自然科学基金项目(编号:81070357;30660066);上海市松江区科学技术攻关项目(编号:14SJGGYY22)

摘　　要：目的:研究腺病毒对原代肝枯否细胞(Kupffer cell,KC)的转染效率和增殖活性影响。方法:分离纯化大鼠肝KC,并以携带绿色荧光蛋白(green fluorescence protein,GFP)基因的腺病毒以不同的感染复数(multiplicity of infection,MOI)转染KC;24 h后,采用荧光显微镜和流式细胞术评估腺病毒的转染效率,以CCK-8比色法检测腺病毒对KC增殖活性的影响。结果 :经统计学处理,当病毒MOI值分别为0、100、300、500、700、900时,荧光显微镜以及流式细胞术测得不同组间GFP表达阳性细胞率均存在明显的差异(均P<0.05)。经CCK8比色法发现,当MOI为300、500、700、900时各组间细胞增殖活性存在明显差异(均P<0.05),而当MOI为0、100、300时各组间细胞增殖活性无明显差异(均P>0.05)。结论 :腺病毒能够有效转染大鼠肝脏KC细胞并表达绿色荧光蛋白。且随着病毒MOI值的增加,腺病毒转染效率逐渐增加。大剂量的病毒对KC增殖活性存在不良影响。Objective To investigate the impacts of adenovirus on the transfection efficiency and proliferative activity of primary Kupffer cells（KCs）. Methods Rat liver KCs were separated and purified by density gradient centrifugation, and was then transfected with adenovirus carrying green fluorescence protein（GFP） gene at different multiplicity of infection（MOI）. After 24h, the transfection efficiency was evaluated by fluorescence microscope and flow cytometry. The proliferative activity of KCs was assessed by colorimetric method.Results The positive percentages of GFP staining cells were statistically different among different doses of adenovirus（MOI 0, 100, 300, 500, 700 and 900） under fluorescence microscopy or by flow cytometry（P0.05 for all comparisons）. The cell proliferative activity had significant differences among MOI 300, 500, 700 and 900（P〈0.05 for all comparisons）, but had no differences among MOI 0, 100 and 300（P〉0.05 for all comparisons） by CCK8 assay. Conclusions KCs can effectively be transfected by GFP adenovirus; and with an increase in virus MOI, the transfection efficiency rises gradually. A higher dose of adenovirus may have a negative effect on cell proliferative.

关 键 词：原代枯否细胞 腺病毒 转染 绿色荧光蛋白 细胞增殖活性 

分 类 号：R373[医药卫生—病原生物学]