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Uric acid drives intestinal barrier dysfunction through TSPO-mediated NLRP3 inflammasome activation

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Abstract

Background and aim

Intestinal epithelial dysfunction is the foundation of various intestinal and extra-intestinal diseases, while the effects and mechanism of uric acid on the intestinal barrier are little known. TSPO has been shown to be related to the generation of ROS and is involved in regulating inflammation, whether uric acid drives intestinal epithelial dysfunction through TSPO-mediated NLRP3 inflammasome activation is unknown.

Methods

UOX gene knockout mouse (UOX-/-) were used for models of hyperuricemia. Fluorescein isothiocyanate (FITC)-labeled dextran was used to assess in vivo intestinal permeability. Serum lipopolysaccharide (LPS) and culture supernatants IL-1β were measured using ELISA Kit. IEC-6 exposed to different concentrations of uric acid was used for in vitro experiment. Protein content and mRNA were assessed using Western blotting and Q-PCR, respectively. Intracellular ROS was determined using flow cytometry and fluorescence microscope. Mitochondrial membrane potential was detected on an immunofluorescence. Small interfering RNA transfection was used to assess the interaction between translocator protein (TSPO) and NLRP3 inflammasome. N-acetyl-L-cysteine (NAC) was used as ROS scavenger.

Results

Our results showed that hyperuricemia mice were characteristic by increased intestinal permeability. Hyperuricemia upregulated TSPO, increased production of ROS and activated NLRP3 inflammasome, which resulted in lower expression of occludin and claudin-1. In vitro, we showed that soluble uric acid alone increased the expression of TSPO, depolarized mitochondrial membrane potential, increased ROS release and activated NLRP3 inflammasome, which further reduced the expression of occludin and claudin-1. Silencing TSPO suppressed NLRP3 inflammasome activation and increased expression of claudin-1 and occludin, which was accompanied by lower levels of ROS. Scavenging ROS also significantly inhibited NLRP3 inflammasome activation without change of TSPO, indicating that TSPO-mediated NLRP3 inflammasome activation was dependent on ROS.

Conclusions

In conclusion, uric acid drives intestinal barrier dysfunction through TSPO-mediated NLRP3 inflammasome.

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Acknowledgements

The study was supported by the National Natural Science Foundation of China (81671625, 81100554, 81901575), Shandong Province Natural Science Fund Project (ZR2018PH010 and ZR2014HM015), Young and Middle-Aged Scientists Research Awards Fund of Shandong Province (BS2012YY003), the Scientific and Technical Development Project of Department of Health of Shandong Province (2011QZ007 and 2016WS0259), a Project of Shandong Province Higher Educational Science and Technology Program (J14LK11) and the Scientific and Technical Development Project of Qingdao (12-1-4-20-jc, 2012-1-3-2-(1)-nsh, 2013-13-008-YY, 2014-1-72 and 17-3-3-15-nsh ).

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Author notes

  1. Qiulan Lv and Daxing Xu contributed equally to this work.

Authors and Affiliations

  1. Medical Research Center, Affiliated Hospital of Qingdao University, No. 167, Wutai Mountain Road, Qingdao, 266003, People’s Republic of China

    Qiulan Lv, Daxing Xu, Jinfeng Ma, Yan Wang, Xiaomin Yang, Peng Zhao, Liang Ma, Zhiyuan Li, Wan Yang, Xiu Liu & Shichao Xing

  2. The Key Laboratory of Mariculture of Chinese Ministry of Education, Ocean University of China, Qingdao, 266003, People’s Republic of China

    Guanpin Yang

  3. School of Cardiovascular Medicine and Science, King’s College London, BHF Centre, London, SE5 9NU, UK

    Shichao Xing

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  1. Qiulan Lv
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Contributions

SX and GY designed the study and revised the manuscript. QL and DX carried out the experiments and wrote the manuscript. YW, WY and XY contributed significantly to analyzing all the data. XL, JM, PZ conducted the mouse experiment. ZL and LM revised the manuscript. All authors approved the final version of the article.

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Correspondence to Shichao Xing.

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Lv, Q., Xu, D., Ma, J. et al. Uric acid drives intestinal barrier dysfunction through TSPO-mediated NLRP3 inflammasome activation. Inflamm. Res. 70, 127–137 (2021). https://doi.org/10.1007/s00011-020-01409-y

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