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pH敏脂质体对反义寡核苷酸抗流感病毒活性的影响
引用本文:陈忠斌,王升启,王弘,朱宝珍,孙志贤.pH敏脂质体对反义寡核苷酸抗流感病毒活性的影响[J].中国生物化学与分子生物学报,1999,15(4):553-557.
作者姓名:陈忠斌  王升启  王弘  朱宝珍  孙志贤
作者单位:军事医学科学院放射医学研究所!北京100850,军事医学科学院放射医学研究所!北京100850,军事医学科学院放射医学研究所!北京100850,军事医学科学院放射医学研究所!北京100850,军事医学科学院放射医学研究所!北京100850
基金项目:国家“863”项目,军队“95”重点项目
摘    要:为了研究具有临床应用前景的 A S O D N 脂质体转运系统,以临床药用大豆磷脂为主要原料制备了p H 敏脂质体,并测定了脂质体体外转染活性、p H 敏特性、细胞毒性和对 A S O D N 抗流感病毒活性的影响 结果发现,批号为 98051903,98051102 和 98051202 的脂质体具有较高转染活性,但只有lipofectin 转染活性的 1/50~1/100当质粒/脂质体( W / W )为 1∶4~1∶8,转染时间为 3~5 h,质粒量为 05 μg,转染后 24~48 h 内检测时转染活性最高 脂质体 98051202 表现明显 p H值依赖溶解红细胞膜特性,而脂质体 98051102 和 98051903 的 p H 敏特性不明显 脂质体细胞毒性明显降低,如 98051903、98051102 和 98051202 的毒性分别是 lipofectin 毒性的 1/16、1/8 和 1/4p H 敏脂质体 98051202 具有促进 A S O D N 抗流感病毒作用,当 A S O D N 浓度为 02 μm ol/ L 时,p H 敏脂质体 98051202 使其抗病毒活性提高 5 倍,但 A S O D N 浓度较高时p H 敏脂质体对 A S O D N抗

关 键 词:pH敏脂质体  反义寡核苷酸  转染  细胞毒性  抗流感病毒活性  
收稿时间:1999-08-20

Effects of pH sensitive Liposomes on Anti influenza Virus Activity of Antisense Oligodeoxynucleotides
CHEN Zhongbin,WANG Shengqi,WANG Hong,ZHU Baozhen,SUN Zhixian.Effects of pH sensitive Liposomes on Anti influenza Virus Activity of Antisense Oligodeoxynucleotides[J].Chinese Journal of Biochemistry and Molecular Biology,1999,15(4):553-557.
Authors:CHEN Zhongbin  WANG Shengqi  WANG Hong  ZHU Baozhen  SUN Zhixian
Institution:(Beijing Institute of Radiation Medicine,Beijing 100850
Abstract:In order to develop an efficient delivery system for antisense oligodeoxynucleotides(ASODN),pH sensitive liposome was prepared and the transfection activity,pH sensitivity, in vitro cytotoxicity and the influence on anti influenza virus of ASODN,No4ODN were assayed Using plasmid pGL3 containing luciferase gene as reporter system,it was found that liposomes 98051903,98051102 and 98051202 showed higher in vitro transfection activity,but their transfection efficiency rates were only 1/50~1/100 of that of lipofectin The highest transfection efficiency could be reached when pGL3 and liposome were mixed in the ratio of 1∶4~1∶8(W/W),with the incubation time of pGL3/liposomes mixture with MDCK cell being 3~5 hours,and the amount of pGL3 0 5 μg,and the assayed time point between 24~48 hours following transfection Liposome 98051202 showed a disruptive property for the red blood cells in a pH dependent manner,whereas liposome 98051903 and 98051102 did not exhibit pH sensitivity The toxicity of prepared liposome was markedly lower than that of lipofectin The lowest toxic concentration of liposome 98051903,98051102 and 98051202 were only 1/16,1/8 and 1/4 of that of lipofectin,respectively Liposome 98051202 showed a significant improved anti influenza virus activity of No4ODN when the concentration of No4ODN was 0 2 μmol/L,but this improvement of antiviral activity of No4ODN declined when the concentration of No4ODN was higher These results indicated that pH sensitive liposomes had the potentials as an efficient drug delivery system for anti influenza virus ASODN with low toxicity
Keywords:pH  sensitive liposome  Antisense oligonucleotides    In vitro  transfection  Cytotoxicity  Anti  influenza virus activity
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