Potential role of recombinant adeno-associated virus human thioredoxin-PR39 in cell and vascular protection against hypoxia

• Published in: Experimental and therapeutic medicine Vol. 9; no. 5; pp. 1605 - 1610
• Main Authors: RUAN, XI-YUN, LIANG, YING-CHUN, DU, BIN, LIN, YOU-TING, GUO, YU-DONG, ZHAO, JING, LI, SHAN, LI, JI-FENG, SUN, QIN-JIAN, DU, YI-FENG
• Format: Journal Article
• Language: English
• Published: Greece D.A. Spandidos 01.05.2015; Spandidos Publications; Spandidos Publications UK Ltd
• Subjects: Adenoviruses; Angiogenesis; Apoptosis; Bioengineering; Care and treatment; Cerebral ischemia; Cloning; Dependoviruses; Enzymes; Fibroblasts; Gene expression; Gene therapy; Health aspects; Hypoxia; Ischemia; Kinases; Methods; Patient outcomes; Peptides; Polymerase chain reaction; Proteins; recombinant adeno-associated virus/human thioredoxin-PR39; Studies; Vascular endothelial growth factor; United States > US; hypoxia; gene therapy; recombinant adeno-associated virus/human thioredoxin-PR39; angiogenesis; ischemia
• ISSN: 1792-0981; 1792-1015
• DOI: 10.3892/etm.2015.2301

The aim of the present study was to successfully construct a recombinant adeno-associated virus (rAAV) vector containing the human thioredoxin (hTRX)-PR39 chimeric gene (rAAV/hTRX-PR39), and verify that the vector was able to maintain a sustained, stable and efficient expression to achieve protein production in the cell. In the present study, a chicken embryo model was utilized to analyze the therapeutical effect of rAAV/hTRX-PR39 in cerebral ischemia diseases. ECV304 cells were transfected with rAAV/hTRX-PR39 and incubated under conditions of 20, 5 and 1% O2. Subsequently, the expression levels of vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor (VEGFR)-1, VEGFR-2, fibroblast growth factor receptor (FGFR)-1 and syndecan-4 were detected by reverse transcription-quantitative polymerase chain reaction. Under hypoxic conditions, the mRNA expression levels of VEGF, VEGFR-1, VEGFR-2, FGFR-1 and syndecan-4 were found to increase in the PR39-transfected group when compared with the control group, while no statistically significant difference was observed between the PR39-transfected group and the control group under conditions of 20% O2. In addition, hTRX-PR39 was shown to increase the density of the vasculature and the survival rate of the chick embryos. Under hypoxic conditions, it was hypothesized that rAAV/hTRX-PR39 was capable of promoting angiogenesis, which may subsequently protect the cells from impairment by hypoxia. In conclusion, rAAV/hTRX-PR39 was demonstrated to promote vascularization and cell survival in hypoxia; thus, rAAV/hTRX-PR39 may have potential for use in therapy targeting cerebral ischemia.