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Wogonin suppresses inflammatory response and maintains intestinal barrier function via TLR4-MyD88-TAK1-mediated NF-κB pathway in vitro

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Abstract

Aims and objective

Wogonin has multiple pharmacological effects, including anti-inflammatory effects. Here, we hypothesize that wogonin can protect intestinal barrier function in lipopolysaccharide (LPS)-induced Caco-2 cells, which is an in vitro model of intestinal inflammation.

Methods

We measured intestinal barrier function in LPS-induced Caco-2 cells by using transepithelial electrical resistance (TEER) and transport of fluorescent markers. A quantitative (q) RT-PCR and immunofluorescent staining analysis was used to detect the expression of tight junction proteins (claudin-1 and ZO-1) in LPS-induced Caco-2 cells. We measured inflammatory molecules in LPS-induced Caco-2 cells using ELISA and qRT-PCR. In addition, the expression of TLR4, MyD88 and TAK1 and their interaction, and NF-κB activity in LPS-induced Caco-2 cells were investigated by western blot analysis and immune-precipitation.

Results

We found that exposing Caco-2 cells to wogonin (10 and 50 μM for 24 h) attenuated the LPS-induced changes in TEER and transport of fluorescent markers. In addition, wogonin suppressed LPS-induced down-regulation of tight junction proteins (claudin-1 and ZO-1). Furthermore, LPS-induced up-regulation of inflammatory mediators, including interleukin (IL)-1β, IL-6 and IL-8, cyclooxygenase-2 (COX-2), inducible nitric oxide synthases (iNOS) were reduced after being pre-treated with wogonin. Moreover, wogonin not only inhibited the expression of TLR4, MyD88 and TAK1 and the interaction between these molecules, but also reduced NF-κB translocation to nucleus and its DNA-binding activity in LPS-induced Caco-2 cells.

Conclusion

Our results suggested that pre-treatment with wogonin could attenuate the TLR4-mediated inflammatory response and maintain intestinal barrier function in LPS-induced Caco-2 cells, thus might be a potential therapy for treating IBD.

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Conflict of interest

The authors declare no conflict of interest.

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Author notes

    Authors and Affiliations

    1. Department of Gastroenterology, The Affiliated Jiangyin Hospital of Southeast University Medical College, Jiangsu, 214400, People’s Republic of China

      Wenping Wang, Tingsong Xia & Xinpu Yu

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    1. Wenping Wang
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    2. Tingsong Xia
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    3. Xinpu Yu
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    Corresponding author

    Correspondence to Xinpu Yu.

    Additional information

    Responsible Editor: John Di Battista.

    W. Wang and T. Xia contributed equally to this work.

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    Wang, W., Xia, T. & Yu, X. Wogonin suppresses inflammatory response and maintains intestinal barrier function via TLR4-MyD88-TAK1-mediated NF-κB pathway in vitro. Inflamm. Res. 64, 423–431 (2015). https://doi.org/10.1007/s00011-015-0822-0

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