肽修饰羧基化多壁碳纳米管基因载体的构建与体外评价

目的:制备一种肽修饰羧基化多壁碳纳米管(peptide modified carboxylated multiwalled carbon nanotubes,MHR)基因载体,考察其对HEK293细胞的转染效率及细胞毒性。方法:将羧化的多壁碳纳米管(MWCNTs)与精氨酸(arginine,R)和组氨酸(histidine,H)组成的短肽(HR)按照一定的质量比反应,通过酰胺键连接得到MHR。采用1 H-NMR、红外光谱以及热重分析对其结构进行鉴定。取纯化后的MHR,用激光粒度测定仪测定粒径和zeta电位,凝胶电泳法测定载体MHR对pEGFP质粒的包裹能力。用MHR/pEGFP纳米复合物与HE...

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Published in药学服务与研究 Vol. 18; no. 1; pp. 11 - 15
Main Author 夏清明;宫春爱;顾芬芬;胡楚玲;高申
Format Journal Article
LanguageChinese
Published 第二军医大学长海医院药学部,上海,200433%上海交通大学医学院附属新华医院药学部,上海,200093%嘉兴市妇幼保健院药学部,浙江嘉兴,314050 2018
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ISSN1671-2838
DOI10.5428/pcar20180103

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Summary:目的:制备一种肽修饰羧基化多壁碳纳米管(peptide modified carboxylated multiwalled carbon nanotubes,MHR)基因载体,考察其对HEK293细胞的转染效率及细胞毒性。方法:将羧化的多壁碳纳米管(MWCNTs)与精氨酸(arginine,R)和组氨酸(histidine,H)组成的短肽(HR)按照一定的质量比反应,通过酰胺键连接得到MHR。采用1 H-NMR、红外光谱以及热重分析对其结构进行鉴定。取纯化后的MHR,用激光粒度测定仪测定粒径和zeta电位,凝胶电泳法测定载体MHR对pEGFP质粒的包裹能力。用MHR/pEGFP纳米复合物与HEK293细胞共同培养,考察细胞摄取情况及相关基因转染情况,并测定MHR和MWCNTs-COOH对DU145和RAW264.7细胞的细胞毒性。结果:通过结构鉴定确定MHR合成成功。在氮磷比(N/P)=20时,HEK293细胞对MHR/pEGFP的摄取及转染效率高于其他N/P值时,其中N/P=20时,RAW264.7细胞对MHR/pGL3复合物的摄取率约为单体pGL3的2.4倍,差异具有统计学意义(P〈0.05)。细胞毒性实验表明,MHR作用于DU145和RAW264.7细胞24和48h后,当MHR浓度达640μg/ml时,DU145和RAW264.7细胞的活性仍然〉80%;而MWCNTs-COOH浓度达320μg/ml时,DU145和RAW264.7细胞活性明显降低;当浓度达640μg/ml时,细胞活性低于20%。结论:MHR有望成为一种高效、低毒的基因载体。
Bibliography:XIA QingMing1 , GONG ChunAi1 , GU FenFen2 , HU ChuLing3 , GAO Shen1(1 .Department of Pharmacy , Changhai Hospital, Second Military Medical University ,Shanghai 200433 ,China?2 .Department of Pharmacy ,Xinhua Hospital Affiliated to School of Medicine ,Shanghai Jiaotong University,Shanghai 200093, China;3 .Department of Pharmacy Jiaxing Maternity and Child Health Care Hospital ,Zhejiang Jiaxing 314050 ,China)
31-1877/R
peptide modified carboxylated multiwalled carbon nanotubes;gene vector;transfection;cytotoxicity
Objective:To prepare peptide modified carboxylated multiwalled carbon nanotubes(MHR)gene vector,and to evaluate its transfection efficiency and cytotoxicity in HEK293 cells.Methods:MHR was obtained by linking the carboxylated multiwalled carbon nanotubes(MWCNTs)with peptide(HR)consisting of arginine(R)and histidine(H)through amide group in a certain ratio of mass.The construction of MHR was characterized by 1 H-NMR,infrared spectrum and thermogravimetric analysis.The particle size and zeta potential of p
ISSN:1671-2838
DOI:10.5428/pcar20180103