Recombinant adenovirus Ad-RUNrf2 reduces paraquat-induced A549 injury
| Autor: | Guangliang Hong, Zhongqiu Lu, J Zheng, X Jiang, Z Chang, Qi-qi Cai, Q Song, Zong-sheng Wu |
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| Rok vydání: | 2012 |
| Předmět: |
Paraquat
Herbicides NF-E2-Related Factor 2 Health Toxicology and Mutagenesis Genetic Vectors Apoptosis General Medicine Biology Toxicology Molecular biology law.invention Adenoviridae chemistry.chemical_compound Oxidative Stress chemistry law Cell Line Tumor Gene expression Recombinant DNA Cytokines Humans |
| Zdroj: | Humanexperimental toxicology. 31(11) |
| ISSN: | 1477-0903 |
| Popis: | Objective: An RU486-inducible recombinant adenovirus-Nrf2 construct (Ad-RUNrf2) was constructed and expressed in H460 cells to determine whether Nrf2 gene expression can be regulated and to observe the effect of the adenovirus Ad-RUNrf2 on inflammatory cytokines, oxidative stress and apoptotic factors that mediate paraquat (PQ)-induced A549 cell injury. Methods: The Nrf2 gene within the RU486 (mifepristone)-inducible system was introduced into an adenovirus vector. A549 cells were transfected with Ad-RUNrf2, and Nrf2 expression was detected using Western blotting and real time-polymerase chain reaction (RT-PCR). RT-PCR, Western blots and enzyme-linked immunosorbent assay were used for observing the effect of RU486-induced Nrf2 expression on the inflammatory cytokines (interleukin-6 (IL-6), IL-10 and tumor necrosis factor-α (TNF-α)), oxidative stress factors (catalase (CAT) and malondialdehyde (MDA)) and apoptosis factors (caspase-3, caspase-9 and cytochrome C) that mediated PQ-induced A549 cell injury. Results: After infection of H460 cells by Ad-RUNrf2, RT-PCR and Western blot analyses showed that Nrf2 expression increased with additional RU486 doses. IL-6 and TNF-α protein and gene expression levels were significantly reduced, and IL-10 protein levels were significantly increased. Although IL-10 expression increased, it remained significantly lower than that of noninduced adenovirus infection and the simple virus exposure group. RU486 induced a significant reduction in MDA expression and increased CAT protein levels. Caspase-9 and caspase-3 protein and gene expression levels decreased in the RU486 induction group ( p < 0.05). Cytochrome C protein levels were not significantly reduced, but its gene expression was significantly decreased ( p < 0.05). Conclusion: Ad-RUNrf2 adenovirus was successfully constructed and can be stably expressed and regulated in cells. Ad-RUNrf2 can reduce PQ-induced inflammation, oxidative stress and apoptosis in A549 cells. |
| Databáze: | OpenAIRE |
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