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Respiratory syncytial virus infection up-regulates TLR7 expression by inducing oxidative stress via the Nrf2/ARE pathway in A549 cells

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Abstract

In order to better understand the early pathways of the pathogenesis of, and immune response to, RSV, herein, we explored the relationship between TLR7 expression and oxidative stress induction following RSV infection in A549 cells. We studied the intervening effects of the Nrf2/ARE pathway agonist butylated hydroxyanisole (BHA) and inhibitor trigonelline (TRI) on TLR7 modulation or oxidative stress induction. For comparison purposes, we set up seven treatment groups in this study, including RSV-treated cells, BHA + RSV-treated cells, TRI + RSV-treated cells, normal cell controls, inactivated RSV controls, BHA controls and TRI controls. We measured changes in TLR7, IL-6, TNF-α mRNA using RT-PCR and IL-6, TNF-α and IL-1β protein using ELISA as well as TLR7, Nrf2 and HO-1 protein using Western blot in A549 cells from the different treatment groups. We also assessed changes in cell proliferation and measured changes in ·OH and NO in A549 cells from the different treatment groups. The results indicate that TLR7 up-regulation is related to RSV infection and the induction of oxidative stress and that TLR7 expression was mediated by the anti-inflammatory effects of Nrf2/ARE pathway inhibitors or agonists. Our experiments may help elucidate the underlying pathology of RSV infection and suggest potential therapeutic targets for drug development and the prevention of RSV-induced human diseases.

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Abbreviations

RSV:

Respiratory syncytial virus

TLR7:

Toll-like receptor 7

pi:

Post infection

BHA:

Butylated hydroxyanisole

TRI:

Trigonelline

Nrf2:

NF-E2-related factor 2

TLRs:

Toll-like receptors

OH:

Hydroxyl radicals

ssRNA:

Single-stranded RNA

COPD:

Chronic obstructive pulmonary disease

AOE:

Antioxidant enzyme

ROS:

Reactive oxygen species

HO-1:

Heme oxygenase-1

HEp-2:

Human laryngeal carcinoma epithelial cells

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Acknowledgements

We thank Ms. **ao-yan Zhang for her technical assistance and LetPub (www.letpub.com) for its linguistic assistance during the preparation of this manuscript.

Funding

This work was funded by the research programs from the National Natural Science Foundation of China (Grant No. 81371797), the program from the Natural Science Foundation of Anhui Province (Grant No. 1308085MH129) and the key project of Natural Science Research of Anhui Education Department (Grant No. KJ2012A152).

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Author notes

  1. Tao Sun, Hai-yang Yu and Chuan-long Zhang contributed equally to this study and should be considered as co-first authors.

Authors and Affiliations

  1. Department of Microbiology, School of Basic Medical Science, Anhui Medical University, Hefei, 230032, People’s Republic of China

    Tao Sun, Hai-yang Yu, Tong-na Zhu & Sheng-hai Huang

  2. School of Life Sciences, Anhui Medical University, Hefei, 230032, People’s Republic of China

    Sheng-hai Huang

  3. Department of Pediatrics, The People’s Hospital of Lu’an City, Lu’an, 237005, Anhui, People’s Republic of China

    Chuan-long Zhang

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  1. Tao Sun
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Correspondence to Sheng-hai Huang.

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Handling Editor: Bert K. Rima.

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Sun, T., Yu, Hy., Zhang, Cl. et al. Respiratory syncytial virus infection up-regulates TLR7 expression by inducing oxidative stress via the Nrf2/ARE pathway in A549 cells. Arch Virol 163, 1209–1217 (2018). https://doi.org/10.1007/s00705-018-3739-4

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