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NF-E2-Related Factor 2 Suppresses Intestinal Fibrosis by Inhibiting Reactive Oxygen Species-Dependent TGF-β1/SMADs Pathway

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Abstract

Background and Aims

This study aimed to evaluate the antifibrotic effects of NF-E2-Related Factor 2 (Nrf2) on intestinal fibrosis. Intestinal fibrosis is a common complication of Crohn’s disease; however, its mechanism of intestinal fibrosis is largely unclear.

Methods

BALB/c mice received 2,4,6-trinitrobenzene sulfonic acid weekly via intrarectal injections to induce chronic fibrotic colitis. They also diet containing received 1% (w/w) tert-butylhydroquinone (tBHQ), which is an agonist of Nrf2. Human intestinal fibroblasts (CCD-18Co cells) were pretreated with tBHQ or si-Nrf2 followed by stimulation with transforming growth factor-β1 (TGF-β1), which transformed the cells into myofibroblasts. The main fibrosis markers such as α-smooth muscle actin, collagen I, tissue inhibitor of metalloproteinase-1, and TGF-β1/SMADs signaling pathway were detected by quantitative real-time RT-PCR, immunohistochemical analysis, and Western blot analysis. Levels of cellular reactive oxygen species (ROS) were detected by dichlorodihydrofluorescein diacetate.

Results

tBHQ suppressed the intestinal fibrosis through the TGF-β1/SMADs signaling pathway in TNBS-induced colitis and CCD-18Co cells. Moreover, Nrf2 knockdown enhanced the TGF-β1-induced differentiation of CCD-18Co cells. ROS significantly increased in TGF-β1-stimulated CCD-18Co cells. Pretreatment with H2O2, the primary component of ROS, was demonstrated to block the effect of tBHQ on reducing the expression of TGF-β1. Moreover, scavenging ROS by N-acetyl cysteine could inhibit the increasing expression of TGF-β1 promoted by Nrf2 knockdown.

Conclusions

The results suggested that Nrf2 suppressed intestinal fibrosis by inhibiting ROS/TGF-β1/SMADs pathway in vivo and in vitro.

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Acknowledgments

The authors thank Experimental center of Shengjing Hospital for technical assistance.

Funding

Funding was provided by Science and Technology Program of Liaoning Province (Grant No. 2013225303).

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Authors and Affiliations

  1. Department of Gastroenterology, Shengjing Hospital of China Medical University, 39 Huaxiang Road, Tiexi District, Shenyang, 110022, Liaoning Province, China

    Yadi Guan, Yue Tan, Jun Yang, Dongxu Wang, Di Pan, Yan Sun & Changqing Zheng

  2. Department of Gastroenterology, The People’s Hospital Liaoning Provincial, 33 Wenyi Road, Shenhe District, Shenyang, 110013, Liaoning Province, China

    Weiyu Liu

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  1. Yadi Guan
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Contributions

CZ and YG conceived and designed the experiments; YG, DP and JY performed the experiments; YS, YG and DW analyzed the data and prepared figures; YG, YT and WL wrote the paper; CZ revised the manuscript for important intellectual content; all authors approval of the final version to be published.

Corresponding author

Correspondence to Changqing Zheng.

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Conflict of interest

The authors declare that they have no conflict of interest. The founding sponsors had no role in study design; in the collection, analysis and interpretation of data; in the writing of the report; and in the decision to submit the article for publication.

Ethical approval

All animal experiments were approved by the institutional care and animal use committee of the China Medical University and conducted in accordance with the National Institute of Health (NIH) Guide for the Care and Use of Laboratory Animals.

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Guan, Y., Tan, Y., Liu, W. et al. NF-E2-Related Factor 2 Suppresses Intestinal Fibrosis by Inhibiting Reactive Oxygen Species-Dependent TGF-β1/SMADs Pathway. Dig Dis Sci 63, 366–380 (2018). https://doi.org/10.1007/s10620-017-4710-z

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