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Deficient Active Transport Activity in Healing Mucosa After Mild Gastric Epithelial Damage

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Abstract

Background

Peptic ulcers recur, suggesting that ulcer healing may leave tissue predisposed to subsequent damage. In mice, we have identified that the regenerated epithelium found after ulcer healing will remain abnormal for months after healing.

Aim

To determine whether healed gastric mucosa has altered epithelial function, as measured by electrophysiologic parameters.

Method

Ulcers were induced in mouse gastric corpus by serosal local application of acetic acid. Thirty days or 8 months after ulcer induction, tissue was mounted in an Ussing chamber. Transepithelial electrophysiologic parameters (short-circuit current, Isc. resistance, R) were compared between the regenerated healed ulcer region and the non-ulcerated contralateral region, in response to luminal hyperosmolar NaCl challenge (0.5 M).

Results

In unperturbed stomach, luminal application of hyperosmolar NaCl transiently dropped Isc followed by gradual recovery over 2 h. Compared to the starting baseline Isc, percent Isc recovery was reduced in 30-day healing mucosa, but not at 8 months. Prior to NaCl challenge, a lower baseline Isc was observed in trefoil factor 2 (TFF2) knockout (KO) versus wild type (WT), with no Isc recovery in either non-ulcerated or healing mucosa of KO. Inhibiting Na/H exchanger (NHE) transport in WT mucosa inhibited Isc recovery in response to luminal challenge. NHE2-KO baseline Isc was reduced versus NHE2-WT. In murine gastric organoids, NHE inhibition slowed recovery of intracellular pH and delayed the repair of photic induced damage.

Conclusion

Healing gastric mucosa has deficient electrophysiological recovery in response to hypertonic NaCl. TFF2 and NHE2 contribute to Isc regulation, and the recovery and healing of transepithelial function.

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Acknowledgments

We thank H.J. Lang, PhD (Aventis Pharma Deutschland) for the generous gift of HOE 694, John Cuppoletti, PhD (University of Cincinnati) for supplying the Ussing chamber, and Chet Closson (University of Cincinnati) for technical assistance with the microscopes. We are very grateful to Timothy C. Wang, MD (Columbia University) for supplying the TFF2-KO, Gary E. Shull, PhD and Roger T. Worrell, PhD (University of Cincinnati) for supplying the NHE2-KO, and Walter Witke, PhD and Jerrold R. Turner, MD, PhD for supplying the HuGE mice.

Funding

This work was supported by the National Institutes of Health (NIH) grant R01DK102551 (M.H.M., E.A.), the University of Cincinnati Research Council Faculty Research Grant (E.A.), Ryuji Ueno Award co-sponsored by the S&R Foundation and American Physiological Society (E.A.), and a VA Merit Award to JDK. This project was also supported in part by the NIH P30 DK078392; Live Microscopy Core and DNA Sequencing and Genotyping Core of the Digestive Disease Research Core Center in Cincinnati.

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Author notes

  1. Izumi Kaji

    Present address: Epithelial Biology Center, Section of Surgical Sciences, Vanderbilt University Medical Center, MRB IV 10435, 2213 Garland Ave., Nashville, TN, 37232, USA

Authors and Affiliations

  1. Department of Pharmacology and Systems Physiology, University of Cincinnati, ML0576, 231 Albert Sabin Way, Cincinnati, OH, 45267, USA

    Andrea L. Matthis, Kristen A. Engevik, Marshall H. Montrose & Eitaro Aihara

  2. Department of Medicine, University of California Los Angeles, Los Angeles, CA, USA

    Izumi Kaji, Yasutada Akiba & Jonathan D. Kaunitz

  3. University of California School of Medicine/Greater Los Angeles VA Healthcare System, Bldg. 114, Suite 217, 11301 Wilshire Blvd., Los Angeles, CA, 90073, USA

    Izumi Kaji & Yasutada Akiba

  4. University of California School of Medicine and Surgery/Greater Los Angeles VA Healthcare System, Bldg. 114, Rm. 217E, 11301 Wilshire Blvd., Los Angeles, CA, 90073, USA

    Jonathan D. Kaunitz

  5. Department of Surgery, University of California Los Angeles, Los Angeles, CA, USA

    Jonathan D. Kaunitz

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  1. Andrea L. Matthis
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Supplementary Fig. 1.

Hypertonic NaCl effect on resistance. Wild-type mouse muscle-stripped gastric corpus was mounted into an Ussing chamber (area = 0.1 cm2) and transepithelial R were measured as described in Methods. Representative time course of percent (%) normalized R after addition of 0 M (black), 0.25 M (green), 0.5 M (blue), or 1.0 M (red) NaCl to luminal nutrient solution. Time zero starts 10 min baseline measurement prior to 5 min (NaCl arrow) exposure to NaCl challenge. At t = 30 min (arrow) shows the time point where change in (Δ) R at 15 min post-NaCl was calculated. (JPEG 176 kb)

Supplementary Fig. 2.

Controls for artifacts during 0.5 M NaCl challenge. After reaching Isc steady state while either a glass coverslip (A) or permeable plastic wrap (B) was mounted in an Ussing chamber, 0.5 M NaCl was added luminally for 5 min (arrow). Outcomes identify Isc changes caused by solution changes that are distinct from tissue active ion transport. (JPEG 170 kb)

Supplementary Fig. 3.

Effect of omeprazole on basal Isc and percent Isc recovery. Wild-type (WT) mouse muscle-stripped gastric corpus was mounted into an Ussing chamber (area = 0.1 cm2). Short-circuit current (Isc) was measured as described in Methods. Results compared never ulcerated tissue (Control, black) or Intact tissue (black) versus the presence of 100 µM omeprazole (OMZ, green-) added a minimum of 30 min prior to 0.5 M NaCl challenge (5 min) to both luminal and serosal bath. (A) Baseline Isc of WT Control without OMZ (−) versus WT Control with OMZ (+), *P < 0.05 (paired two-tailed t test). (B) Percent (%) Isc recovery for WT Intact (from Fig. 2c) without OMZ (−) versus WT Control with OMZ (+). (C) Percent (%) Isc recovery for TFF2-KO Intact (from Fig. 4b) without OMZ (−) versus TFF2-KO Control with OMZ (+). Mean ± SEM (n = 4 WT Control or WT Intact or TFF2 Control, n = 5 TFF2-KO Intact). (JPEG 178 kb)

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Matthis, A.L., Kaji, I., Engevik, K.A. et al. Deficient Active Transport Activity in Healing Mucosa After Mild Gastric Epithelial Damage. Dig Dis Sci 65, 119–131 (2020). https://doi.org/10.1007/s10620-019-05825-x

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