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Enteropathogenic Escherichia coli Infection Inhibits Intestinal Ascorbic Acid Uptake via Dysregulation of Its Transporter Expression

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Abstract

Background

Enteropathogenic Escherichia coli (EPEC) infection causes prolonged, watery diarrhea leading to morbidity and mortality. Although EPEC infection impacts nutrient transporter function and expression in intestinal epithelial cells, the effects of EPEC infection on intestinal absorption of ascorbic acid (AA) have not yet been investigated.

Aims

To investigate the effect of EPEC infection on intestinal AA uptake process and expression of both AA transporters.

Methods

We used two experimental models: human-derived intestinal epithelial Caco-2 cells and mice. 14C-AA uptake assay, Western blot, RT-qPCR, and promoter assay were performed.

Results

EPEC (WT) as well as ΔespF and ΔespG/G2 mutant-infected Caco-2 cells showed markedly inhibited AA uptake, while other mutants (ΔescN, ΔespA, ΔespB, and ΔespD) did not affect AA uptake. Infection also reduced protein and mRNA expression levels for both hSVCT1 and hSVCT2. EPEC-infected mice showed marked inhibitory effect on AA uptake and decreased protein and mRNA expression levels for both mSVCT1 and mSVCT2 in jejunum and colon. MicroRNA regulators of SVCT1 and SVCT2 (miR103a, miR141, and miR200a) were upregulated significantly upon EPEC infection in both Caco-2 and mouse jejunum and colon. In addition, expression of the accessory protein glyoxalate reductase/hydroxypyruvate reductase (GRHPR), which regulates SVCT1 function, was markedly decreased by EPEC infection in both models.

Conclusions

These findings suggest that EPEC infection causes inhibition in AA uptake through a multifactorial dysregulation of SVCT1 and SVCT2 expression in intestinal epithelial cells.

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Acknowledgments

The current study was supported by the National Institutes of Health grants DK107474 (VSS), DK56061 (HMS), AA018071 (HMS), and GM088790 (JSM), MH108154 (MGG), as well as a grant from the Department of Veterans Affairs (HMS). We thank Dr. Gail Hecht (Loyola University, Chicago) for providing EPEC mutant strains.

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Authors and Affiliations

  1. Department of Medicine, University of California Irvine, Irvine, CA, 92697, USA

    Christopher W. Heskett, Trevor Teafatiller, Hamid M. Said & Veedamali S. Subramanian

  2. Department of Physiology and Biophysics, University of California Irvine, Irvine, CA, 92697, USA

    Hamid M. Said

  3. Veterans Affairs Medical Center, Long Beach, CA, 90822, USA

    Hamid M. Said & Veedamali S. Subramanian

  4. Department of Anatomy, Physiology and Cell Biology, School of Veterinary Medicine, University of California Davis, Davis, CA, 95616, USA

    Carly Hennessey & Melanie G. Gareau

  5. Department of Cell Biology, Neurobiology and Anatomy, Medical College of Wisconsin, Milwaukee, WI, 53226, USA

    Jonathan S. Marchant

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  1. Christopher W. Heskett
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Correspondence to Veedamali S. Subramanian.

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Heskett, C.W., Teafatiller, T., Hennessey, C. et al. Enteropathogenic Escherichia coli Infection Inhibits Intestinal Ascorbic Acid Uptake via Dysregulation of Its Transporter Expression. Dig Dis Sci 66, 2250–2260 (2021). https://doi.org/10.1007/s10620-020-06389-x

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