中国生物医学工程学报
中國生物醫學工程學報
중국생물의학공정학보
CHINESE JOURNAL OF BIOMEDICAL ENGINEERING
2010年
1期
129-136
,共8页
温玉婷%潘仕荣%郭振寰%王持%曾昕%吴红梅%冯敏
溫玉婷%潘仕榮%郭振寰%王持%曾昕%吳紅梅%馮敏
온옥정%반사영%곽진환%왕지%증흔%오홍매%풍민
聚酰胺-胺树枝状高分子%组氨酸%血清%基因转染%基因载体
聚酰胺-胺樹枝狀高分子%組氨痠%血清%基因轉染%基因載體
취선알-알수지상고분자%조안산%혈청%기인전염%기인재체
PAMAM%histidine%serum%gene transfection%gene vector
组氨酸的咪唑环由于其独特的质子化机理而受到了广泛的关注.研究表明,组氨酸修饰的基因载体可对抗血清对基因转染的影响,提高载体在血清中的基因转染效率.本研究利用氨解反应将组氨酸接枝到聚酰胺-胺树枝状高分子(PAMAM)的表面,制备了一种新型的PAMAM衍生物-组氨酸修饰的聚酰胺-胺树枝状高分子(His-PAMAM G4).利用~1H NMR对His-PAMAM G4进行表征,证明一个PAMAM G4分子上接枝37个组氨酸分子.对His-PAMAM G4/DNA复合物进行结合DNA能力、粒径、表面电位、粒子形态等理化性能的表征,证明该新型聚合物对DNA分子具有良好的压缩能力.细胞活性检测结果表明,His-PAMAM G4对Bel 7402和Hela细胞的毒性均显著低于未经修饰的PAMAM G4.His-PAMAM G4在血清中的转染效率与PAMAM G4相比大幅度提高,也显著高于阳离子聚合物PEI 25k和市售商品阳离子脂质体Lipofectamine.因此,His-PAMAM G4有望成为一种高效、安全、可在体内应用的非病毒基因载体.
組氨痠的咪唑環由于其獨特的質子化機理而受到瞭廣汎的關註.研究錶明,組氨痠脩飾的基因載體可對抗血清對基因轉染的影響,提高載體在血清中的基因轉染效率.本研究利用氨解反應將組氨痠接枝到聚酰胺-胺樹枝狀高分子(PAMAM)的錶麵,製備瞭一種新型的PAMAM衍生物-組氨痠脩飾的聚酰胺-胺樹枝狀高分子(His-PAMAM G4).利用~1H NMR對His-PAMAM G4進行錶徵,證明一箇PAMAM G4分子上接枝37箇組氨痠分子.對His-PAMAM G4/DNA複閤物進行結閤DNA能力、粒徑、錶麵電位、粒子形態等理化性能的錶徵,證明該新型聚閤物對DNA分子具有良好的壓縮能力.細胞活性檢測結果錶明,His-PAMAM G4對Bel 7402和Hela細胞的毒性均顯著低于未經脩飾的PAMAM G4.His-PAMAM G4在血清中的轉染效率與PAMAM G4相比大幅度提高,也顯著高于暘離子聚閤物PEI 25k和市售商品暘離子脂質體Lipofectamine.因此,His-PAMAM G4有望成為一種高效、安全、可在體內應用的非病毒基因載體.
조안산적미서배유우기독특적질자화궤리이수도료엄범적관주.연구표명,조안산수식적기인재체가대항혈청대기인전염적영향,제고재체재혈청중적기인전염효솔.본연구이용안해반응장조안산접지도취선알-알수지상고분자(PAMAM)적표면,제비료일충신형적PAMAM연생물-조안산수식적취선알-알수지상고분자(His-PAMAM G4).이용~1H NMR대His-PAMAM G4진행표정,증명일개PAMAM G4분자상접지37개조안산분자.대His-PAMAM G4/DNA복합물진행결합DNA능력、립경、표면전위、입자형태등이화성능적표정,증명해신형취합물대DNA분자구유량호적압축능력.세포활성검측결과표명,His-PAMAM G4대Bel 7402화Hela세포적독성균현저저우미경수식적PAMAM G4.His-PAMAM G4재혈청중적전염효솔여PAMAM G4상비대폭도제고,야현저고우양리자취합물PEI 25k화시수상품양리자지질체Lipofectamine.인차,His-PAMAM G4유망성위일충고효、안전、가재체내응용적비병독기인재체.
The unique protonation mechanism of imidazole in histidine molecules attracts wide research attention in recent years. It has been reported by literatures that the impact of serum to the transfection efficiency could be reduced by modifying gene vectors with histidine to obtain enhanced transfection efficiency. In this work, histidine was conjugated to the surface of PAMAM G4 using aminolysis reaction to construct a PAMAM derivative (His-PAMAM G4) as a new gene vector. The chemistry of His-PAMAM G4 was characterized using 1H NMR, which showed that one PAMAM was linked with 37 histidine. The DNA binding ability, particle size, zeta potential, and the morphology of the His-PAMAM G4/DNA complexe were investigated. Experimental results revealed that the His-PAMAM G4 could sufficiently condense DNA. MTT assay indicated that the cytotoxicity of the His-PAMAM G4 was obviously lower than PAMAM G4 control to Bel 7402 cells and Hela cells. The transfection efficiency of the His-PAMAM G4 detected by GFP flow cytometry was significantly enhanced in reference with that of PAMAM G4 control in serum. Therefore, the His-PAMAM G4 holds a potential of nonviral gene vectors in gene therapy in vivo.