Inhibition of Neointima Hyperplasia, Inflammation, and Reactive Oxygen Species in Balloon-Injured Arteries by HVJ Envelo
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ORIGINAL ARTICLE
Inhibition of Neointima Hyperplasia, Inflammation, and Reactive Oxygen Species in Balloon-Injured Arteries by HVJ Envelope Vector-Mediated Delivery of Superoxide Dismutase Gene Shoa-Lin Lin 1,2 & Jwu-Lai Yeh 3,4,5 & Pei-Chia Tsai 6 & Tsung-Hsien Chang 7,8 & Wei-Chun Huang 2,6 & Song-Tay Lee 9 & Michael Wassler 10 & Yong-Jian Geng 10 & Erna Sulistyowati 3,11 Received: 11 December 2017 / Revised: 18 August 2018 / Accepted: 22 August 2018 # The Author(s) 2018
Abstract Extracellular superoxide dismutase (EC-SOD) has been implicated in regulation of vascular function but its underlying molecular mechanism is largely unknown. These two-step experiments investigate whether hemagglutinating virus of Japan envelope (HVJ-E) vector-mediated EC-SOD gene delivery might protect against neointima formation, vascular inflammation, and reactive oxygen species (ROS) generation, and also explore cell growth signaling pathways. The first in-vitro experiment was performed to assess the transfection efficacy and safety of HVJ-E compared to lipofectamine®. Results revealed that HVJ-E has higher transfection efficiency and lower cytotoxicity than those of lipofectamine®. Another in-vivo study initially used balloon denudation to rat carotid artery, then delivered EC-SOD cDNA through the vector of HVJ-E. Arterial section with H&E staining from the animals 14 days after balloon injury showed a significant reduction of intima-to-media area ratio in EC-SOD transfected arteries when compared with control (empty vector-transfected arteries) (p < 0.05). Arterial tissue with EC-SOD gene delivery also exhibited lower levels of ROS, as assessed by fluorescent microphotography with dihydroethidium staining. Quantitative RT-PCR revealed that EC-SOD gene delivery significantly diminished mRNA expression of tumor necrosis factor (TNF)-α and interleukin (IL)-1β (p < 0.05 in all comparisons). An immunoblotting assay from vascular smooth muscle cell (VSMC) cultures showed that the EC-SOD transfected group attenuated the activation of MEK1/2, ERK1/2, and Akt signaling significantly. In conclusion, EC-SOD overexpression by HVJ-E vector inhibits neointima hyperplasia, inflammation, and ROS level triggered by balloon injury. The modulation of cell growth-signaling pathways by EC-SOD in VSMCs might play an important role in these inhibitory effects. Keywords Gene therapy . Reactive oxygen species . Restenosis . Superoxide dismutase
Shoa-Lin Lin and Jwu-Lai Yeh contributed equally to this work. * Shoa-Lin Lin [email protected] 1
Intensive Care Unit, Yuan’s General Hospital, 162, Cheng-Kung First Road, Lingya District, Kaohsiung 80249, Taiwan
2
School of Medicine, National Yang-Ming University, Taipei, Taiwan
3
Department of Pharmacology and Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan
4
Department of Medical Research, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan
5
Department of Marine Biotechnology and Resources, National Sun Yat-sen University, Kaohsiung, Taiwa
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