摘要
目的制备携Pik3cb shRNA的超顺磁性氧化铁纳米微泡,研究其物理特性,并通过在体实验评价其抑制血管平滑肌细胞增殖的效果。材料与方法反相微乳法制备SPIO纳米微泡,利用静电吸附效应负载Pik3cb shRNA后得到携Pik3cb shRNA的SPIO纳米微泡(SPIO-shPik3cb),检测其粒径大小及形貌,琼脂糖凝胶电泳检测其结合质粒的能力,小动物超声仪检测体外超声成像效果和靶向性。成功建立大鼠颈动脉球囊损伤模型,分组进行转染实验。HE染色观察新生血管内膜的增生情况,测量并比较内膜与中膜面积(I/M)比值。Western blot检测磷酸化Akt相关蛋白的表达。结果SPIO-shPik3cb负载质粒前后的平均粒径分别为(143.60±18.27)nm和(151.45±11.18)nm,可以紧密结合质粒,具有一定的体外超声成像效果和靶向性。HE染色结果显示SPIO-shPik3cb组转染实验14 d后的I/M比值为0.48±0.08,低于建模对照组的0.96±0.12和Pik3cb shRNA组的0.66±0.07,能显著抑制新生内膜的增生(t=4.872,P<0.05)。Western blot结果显示SPIO-shPik3cb能明显下调磷酸化Akt蛋白的表达(P<0.05)。结论SPIO-shPik3cb适用于基因药物的递送,在磁场和靶向超声的双重作用下,可以通过下调磷酸化Akt蛋白的表达,有效抑制血管平滑肌细胞增殖。
Purpose To prepare superparamagnetic iron oxide(SPIO)nanoparticles loaded with Pik3cb shRNA,study their physical properties,and evaluate their effect on inhibiting the proliferation of vascular smooth muscle cells by in vivo experiments.Materials and Methods SPIO magnetic nanoparticles were prepared by reversed-phase microemulsion method,and the SPIO-shPik3cb were loaded with Pik3cb shRNA by electrostatic adsorption effect,their particle size and morphology were examined.The imaging effect and targeting were examined by small animal ultrasonography.A rat carotid artery balloon injury model was established and grouped for transfection experiments.I/M ratio of intima-media to intima-media area was measured by HE staining and compared.The expression of phosphorylated Akt was measured by Western bolt.Results The mean particle size of SPIO-shPik3cb before and after plasmid loading were(143.60±18.27)nm and(151.45±11.18)nm,respectively,which could tightly bind the plasmid and had certain in vitro ultrasound imaging effect and targeting.HE staining showed that the I/M ratio of SPIO-shPik3cb group after 14 d of injury was 0.48±0.08.The transfection efficiency was lower than that of the control group(0.96±0.12)and the Pik3cb shRNA group(0.66±0.07),which significantly inhibited the proliferation of endothelium(t=4.872,P<0.05).Western blot results showed that SPIO-shPik3cb significantly downregulated the expression of phosphorylated Akt(P<0.05).Conclusion SPIO-shPik3cb is suitable for gene drug delivery and can effectively inhibit the proliferation of vascular smooth muscle cells by down-regulating the expression of phosphorylated Akt protein under the dual action of magnetic field and targeted ultrasound.
作者
王宇豪
马小五
方玲玲
李林
李妙
张平洋
WANG Yuhao;MA Xiaowu;FANG Lingling;LI Lin;LI Miao;ZHANG Pingyang(Department of Cardiovascular Ultrasound,Nanjing First Hospital,Nanjing Medical University,Nanjing 210006,China)
出处
《中国医学影像学杂志》
CSCD
北大核心
2022年第6期529-534,共6页
Chinese Journal of Medical Imaging
基金
南京市医学科技发展重点项目(ZKX17026)。
关键词
超声检查
超顺磁性氧化铁
靶向微泡
血管平滑肌细胞
Ultrasonography
Superparamagnetic iron oxide
Target nanoparticle
Vascular smooth muscle cells
作者简介
通信作者:张平洋,zhpy28@126.com。
