中国社区医师Issue(24)：5-6,8,3.DOI:10.3969/j.issn.1007-614x.2014.24.1
阳离子磷酸胆碱聚合物载AT1受体反义寡核苷酸转染大鼠心肌成纤维细胞的研究
Study on cationic phosphorylcholine polymer load AT1 receptor of antisense oligonucleotide to transfected rat myocardium fibroblasts
孙敏 1鲁永织 2程莹 1王志荣 1张卓琦1
作者信息
- 1. 221002徐州医学院附属医院心内科 江苏徐州
- 2. 266011山东省青岛市市立医院肿瘤二科
- 折叠
摘要
Abstract
Objective:To investigate the impact of novel cationic phosphorylcholine polymer of MPC30-DEA70 load AT1 receptor of antisense oligonucleotide(AS-ODN) on the expression of AT1 receptor and type I collagen in cultured rat myocardial. Methods:MTT assay for detection of MPC and cardiac fibroblasts cell compatibility;application of laser scanning confocal microscope(CLAM) to detect complex in the cell,flow cytometry(FCM) detected transfection efficiency and fluorescence intensity, the expression of Western blot detection of AT1 receptor protein and collagen I.Results:When the concentration of MPC>40 μg/ml cytotoxicity will obviously appeared,under the confocal laser scanning microscopy,most of gene complexes marked by FAM distributed in nucleus around,a small amount of them distributed in the nucleus.Flow cytometry analysis revealed that with N/P ratio increased the efficiency of transfection and fluorescence intensity increased obviously.The Western blot results showed that expression of intracellular AT1 receptor and type I collagen was reduced accordingly after transfection complexes.Conclusion:MPC30-DEA70 can carry AT1-R AS-ODN in cultured rat cardiac fibroblasts with high transfection rate in the safe concentration range,and inhibit the expression of AT1R and type I collagen.So then we can infer that cationic phosphorylcholine polymer can effectively load antisense oligonucleotide(AS-ODN),targeted inhibition of gene expression.This research will provide certain theoretical basis for study on the novel non viral gene vector.关键词
磷酸胆碱聚合物/AS-ODN/AT1受体/非病毒基因载体Key words
Phosphorylcholine polymer/AS-ODN/AT1 receptor/Non viral gene vector引用本文复制引用
孙敏,鲁永织,程莹,王志荣,张卓琦..阳离子磷酸胆碱聚合物载AT1受体反义寡核苷酸转染大鼠心肌成纤维细胞的研究[J].中国社区医师,2014,(24):5-6,8,3.
