Thioredoxin gene recombination and expression in Neuro-2A cells

doi:10.3969/j.issn.2095-4344.2013.11.019

Abstract

Abstract:

BACKGROUND: Studies on antioxidant genes Thioredoxin (TRX) have attracted more attention gradually, but relevant studies from the aspect of gene therapy are fewer.
OBJECTIVE: To study the protective effect of cells expressing the corresponding proteins in TRX transfected Neuro-2A cells and to analyze the possible mechanism underlying the protective effect.
METHODS: Plasmids PIRES2-EGFP-TRX were used to transfect Neuro-2A cells and to establish a cell line which could stably express TRX proteins identified by reverse transcription-PCR. Different concentrations of hydrogen peroxide were used to treat normal cells and transfected cells for the establishment of oxidative stress models. Cell morphology, cell survival, glutathione concentration and DNA strand breaks situation were observed after oxidative damage.
RESULTS AND CONCLUSION: Normal cells and transfected cells were both damaged by hydrogen peroxide; however, the transfected cells were superior to the normal cells in cell damage, cell survival,
intracellular glutathione level and degree of DNA strand breaks. These findings indicate that TRX gene in Neuro-2A cells can be reconstructed and expressed successfully, which plays a certain protective. This effect may be achieved through scavenging oxygen free radicals, maintaining intracellular glutathione levels, thereby protecting cellular DNA against oxidative damage.

Key words: tissue construction, cytology experiment in tissue construction, Thioredoxin, transfection, Neuro-2A cells, oxidative stress, protection, oxygen free radicals, glutathione, stress models, single cell gel electrophoresis, tissue construction photographs-containing paper

CLC Number:

R318

Wang Long, Lü Guo-feng. Thioredoxin gene recombination and expression in Neuro-2A cells[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(11): 2026-2031.

Figures/Tables 7

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