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Recombinant adenovirus Ad-RUNrf2 reduces paraquat-induced A549 injury.
Hum Exp Toxicol. 2012 Nov; 31(11):1102-12.HE

Abstract

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.

Authors+Show Affiliations

Emergency Department, The First Affiliated Hospital of Wenzhou Medical College, Wenzhou, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

22736252

Citation

Cai, Q, et al. "Recombinant Adenovirus Ad-RUNrf2 Reduces Paraquat-induced A549 Injury." Human & Experimental Toxicology, vol. 31, no. 11, 2012, pp. 1102-12.
Cai Q, Lu Z, Hong G, et al. Recombinant adenovirus Ad-RUNrf2 reduces paraquat-induced A549 injury. Hum Exp Toxicol. 2012;31(11):1102-12.
Cai, Q., Lu, Z., Hong, G., Jiang, X., Wu, Z., Zheng, J., Song, Q., & Chang, Z. (2012). Recombinant adenovirus Ad-RUNrf2 reduces paraquat-induced A549 injury. Human & Experimental Toxicology, 31(11), 1102-12. https://doi.org/10.1177/0960327112450902
Cai Q, et al. Recombinant Adenovirus Ad-RUNrf2 Reduces Paraquat-induced A549 Injury. Hum Exp Toxicol. 2012;31(11):1102-12. PubMed PMID: 22736252.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Recombinant adenovirus Ad-RUNrf2 reduces paraquat-induced A549 injury. AU - Cai,Q, AU - Lu,Z, AU - Hong,G, AU - Jiang,X, AU - Wu,Z, AU - Zheng,J, AU - Song,Q, AU - Chang,Z, Y1 - 2012/06/26/ PY - 2012/6/28/entrez PY - 2012/6/28/pubmed PY - 2013/5/1/medline SP - 1102 EP - 12 JF - Human & experimental toxicology JO - Hum Exp Toxicol VL - 31 IS - 11 N2 - 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. SN - 1477-0903 UR - https://www.unboundmedicine.com/medline/citation/22736252/Recombinant_adenovirus_Ad_RUNrf2_reduces_paraquat_induced_A549_injury_ L2 - https://journals.sagepub.com/doi/10.1177/0960327112450902?url_ver=Z39.88-2003&amp;rfr_id=ori:rid:crossref.org&amp;rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -