Synergistic effects of ultrasound-targeted microbubble destruction and TAT peptide on gene transfection: An experimental study in vitro and in vivo
Synergistic effects of ultrasound-targeted microbubble destruction and TAT peptide on gene transfection: An experimental study in vitro and in vivo
复制标题
超声靶向微泡破坏和 TAT 肽对基因转染的协同效应：体外和体内实验研究
DOI：
10.1016/j.jconrel.2013.06.005
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发表时间：
2013-09-28
影响因子：
10.8
通讯作者：
Wang, Zhigang
中科院分区：
文献类型：
--
作者：
Zhou, Zhiyi;Zhang, Ping;Wang, Zhigang
细胞渗透性肽（CPP）和超声靶向微泡破坏（UTMD）在基因递送方面具有巨大的潜力。然而，由于CPPs的非特异性和UTMD的低转染效率，它们的应用受到限制。本研究通过脂质微泡将达特肽（TATp）和肝细胞生长因子（HGF）基因包裹在脂质微泡中，使TATp不被酶切，HGF基因不被降解，从而构建了一种“智能”基因传递系统。这种新策略具有UTMD和TATp的协同作用。本研究探讨了UTMD和TATp联合介导的HGF基因转染的有效性和安全性。MTT法和流式细胞仪分析结果表明，UTMD和TATp联合应用可提高HUVECs中HGF基因的表达，且对细胞活力无明显影响。在大鼠心肌梗死模型中，UTMD和TATp联合应用可使心肌组织中HGF的蛋白和mRNA表达最高，组织学结果显示UTMD和TATp联合应用可增强心肌微血管形成，减轻心肌纤维化。结论：联合应用UTMD和TATp是一种安全有效的基因转移技术。(C)2013年爱思唯尔B。版权所有© 2016
Cell-permeable peptides (CPPs) and ultrasound-targeted microbubble destruction (UTMD) have tremendous potential for gene delivery. However, their applications are limited due to nonspecificity of CPPs and low transfection efficiency of UTMD. Here, we developed a 'smart' gene delivery system by encapsulating TAT peptide (TATp) and hepatocyte growth factor (HGF) gene within lipid microbubbles, in which TATp was protected from being enzymatically cleaved and HGF gene was protected from degradation. This new strategy had synergistic effects of UTMD and TATp on gene transfection. We investigated the efficacy and safety of HGF gene transfection mediated by the combination of UTMD and TATp in vitro and in vivo. The results from MTT assay and flow cytometry analyses indicated that the combination of UTMD and TATp could enhance HGF gene expression in HUVECs without any significant side effect on cell viability. In rat myocardial infarction models, we demonstrated that the protein and mRNA expressions of HGF in myocardium caused by the combination of UTMD and TATp were the highest.Histopathological findings demonstrated that the combination of UTMD and TATp enhanced myocardial microvasculature and ameliorated myocardial fibrosis. In conclusion, the combination of UTMD and TATp might be a safe and efficient technique for gene delivery. (C) 2013 Elsevier B. V. All rights reserved.