Inhibition of Neointima Hyperplasia, Inflammation, and Reactive Oxygen Species in Balloon-Injured Arteries by HVJ Envelope Vector-Mediated Delivery of Superoxide Dismutase Gene

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.

Inflammatory responses induced by Helicobacter pylori on the carcinogenesis of gastric epithelial GES‑1 cells

“…The cDNA was prepared using M-MLV reverse transcriptase (Promega Corporation, Madison, WI, USA). A mixture consisting of RNA (500 ng/µl; 2 µl), 5X M-MLV RT buffer (12 µl), dNTP mixes (2.5 mM; 6 µl), RNase inhibitor (30 U/µl; 1 µl), M-MLV RT (5 U/µl; 4 µl), Oligo dT (18) primer (500 ng/µl; 3 µl) and diethyl pyrocarbonate water (17 µl) was cultured at 37˚C for 1 h. RT-qPCR was performed on an Eppendorf PCR system (Eppendorf, Hamburg, Germany). cDNA (1 µl), forward and reverse primers (each 0.5 µl), SYBR ® Premix ExTaq (8 µl; Takara Biotechnology Co., Ltd., Dalian, China) and ddH 2 O (9 µl) were added to the mix, followed by 32 cycles of amplification as follows: Denaturation at 94˚C for 2 min; 32 cycles of 94˚C for 20 sec, 55˚C for 35 sec, 72˚C for 25 sec; and extension at 72˚C for 5 min.…”

“…The host cells employ reactive oxygen species and nitric oxide to eliminate an invading pathogen, but this reaction mechanism could also regulate the inflammatory response of host cells (18). A total of 2 major enzymes (iNOS and COX-2) involved in the metabolism of HP-infected GES-1 cells (Fig.…”

The Effects of Pro-Inflammatory and Anti-Inflammatory Agents for the Suppression of Intimal Hyperplasia: An Evidence-Based Review

“…Inflammation within the vascular wall is a hurdle, and manipulating the inflammatory cytokines and chemokines is possibly the key piece in solving the puzzle. Other than that, research related to inhibition of intimal hyperplasia has also been extensively done using various approaches, which utilized gene therapy and vector-mediated gene delivery [ 49 , 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 , 60 , 61 , 62 , 63 , 64 ], anti-inflammatory and anti-proliferative drugs [ 57 ], compounds with anti-inflammatory actions such as plants [ 55 , 65 ], herbal medications [ 50 ], marine sources [ 52 ] and polyunsaturated fatty acids [ 56 ].…”

Bilirubin Nanomedicines for the Treatment of Reactive Oxygen Species (ROS)-Mediated Diseases

“…31 In addition, monoamine oxidase 32 and cytochrome b5 reductase 33 were also reported to have physiological activities to promote the formation of ROS. There is an endogenous antioxidative system in organisms, which mainly consists of superoxide dismutase (SOD) (scavenges O 2 • − ), 6 glutathione peroxidase (GPx) (eliminates H 2 O 2 and lipid peroxide), 8 and catalase (catalyzes the decomposition of H 2 O 2 ). 7 Therefore, enhancing the activity of these enzymes and accelerating the transformation and deactivation of free radicals can protect the cells from the damaging effects of excess ROS.…”

“…Nowadays, one of the effective strategies is to use antioxidants to scavenge redundant ROS. Antioxidants are categorized as nonenzymatic and enzymatic antioxidants. , Enzymatic antioxidants mainly include superoxide dismutase, catalase, and glutathione peroxidase, whereas nonenzymatic antioxidants mainly consist of polyphenols (anthocyanins, lycopene, resveratrol, and tea polyphenols), vitamins (vitamins E, C, and A, and carotene complex), and other kinds of natural products. , Among them, inartificial nonenzymatic antioxidants have increasingly attracted great interest from supporters and pharmaceutical companies due to their abundant sources, significant antioxidant activities, and rare side effects. − …”