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Chronic Estrogen Deficiency Causes Gastroparesis by Altering Neuronal Nitric Oxide Synthase Function

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Abstract

Background

Gastroparesis affects predominantly females; however, the biological basis for this gender bias is completely unknown. Several lines of evidence suggest that nitrergic dependent stomach motility function is reduced in diabetic gastroparesis and that nNOS is estrogen-regulated.

Aims

The purpose of this study was to investigate whether reduced levels of estradiol-17β (E2) down-regulates tetrahydrobiopterin (BH4, a cofactor for nNOS dimerization and enzyme activity) biosynthesis and therefore nNOS mediated gastric motility would be impaired in a mouse model of chronic estrogen deficiency, follicle stimulating hormone receptor knock-out female mice (FORKO).

Methods

In-bred 12-week-old female FORKO mice were obtained from our FORKO breeding colony. Gastric emptying was measured in overnight fasting mice. Nitrergic relaxation (AUC/mg tissue) was measured at 2 Hz through electric field stimulation using gastric antrum strips prepared from WT and FORKO mice. Protein expression for nNOSα, BH4 biosynthesis enzymes (GCH-1, DHFR) and estrogen receptors (α, β) were measured in gastric antrum by western blotting. Levels of BH4 and oxidized BH2, B biopterin levels were determined by HPLC.

Results

In FORKO, compared to wild type (WT) stomachs we indentified (1) reduced (%) gastric emptying (64 ± 2.5 vs. 77.6 ± 0.88), (2) greater reduction in nitregic relaxation (−0.13 ± 0.012 vs. −0.28 ± 0.012), (3) increased nNOS dimerization (0.48 ± 0.02 vs. 0.34 ± 0.05), (4) decreased NO release whether measured at 24 h (0.6 ± 0.04 vs. 1.7 ± 0.22, p < 0.05) or at 48 h (3.4 ± 0.26 vs. 5.0 ± 0.15, p < 0.05) of incubation, (5) decreased GCH-1 (1.9 ± 0.06 vs. 2.3 ± 0.04), DHFR (1.8 ± 0.14 vs. 2.4 ± 0.07) and ERα (2.7 ± 0.4 vs. 3.9 ± 0.4) and (6) increased oxidized biopterin levels and decreased ratio of BH4 versus BH2 + B.

Conclusion

We conclude that chronic estrogen deficiency negatively modifies the function of both BH4 and nNOS thereby contributing to the development of gastroparesis in a FORKO mouse model.

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Acknowledgments

We thank Dr. Ayman Al-Hendy for the FORKO and SVE129 mice, and Dr. Diana Marver for review of the manuscript. Financial support for the original research was provided by NIH-NIDDK R21DKO76704 (PG), P60DK020593 pilot project funds (PG), and RCMI G12RR03032 (PG) start-up funds at Meharry Medical College, Nashville, TN, USA.

Conflict of interest

Dr. Gangula has filed a patent application for the use of BH4 in gastroparesis subjects through the University of Texas Medical Branch, Galveston, TX.

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Authors and Affiliations

  1. Department of Physiology, Meharry Medical College, 1005 Dr. D.B. Todd Jr. Blvd, Nashville, TN, 37208, USA

    K. Ravella & P. R. Gangula

  2. Center for Women’s Health Research, Meharry Medical College, Nashville, TN, 37208, USA

    A. Al-Hendy, C. Sharan & P. R. Gangula

  3. Department of Obstetrics and Gynecology, Meharry Medical College, Nashville, TN, 37208, USA

    A. Al-Hendy & C. Sharan

  4. Department of Cardiovascular Medicine, John Radcliffe Hospital, University of Oxford, Oxford, OX3 9DU, UK

    A. B. Hale & K. M. Channon

  5. Division of Gastroenterology and Hepatology, Emory University Medical Center, Atlanta, GA, USA

    S. Srinivasan

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  1. K. Ravella
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  3. C. Sharan
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  7. P. R. Gangula
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Correspondence to P. R. Gangula.

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Ravella, K., Al-Hendy, A., Sharan, C. et al. Chronic Estrogen Deficiency Causes Gastroparesis by Altering Neuronal Nitric Oxide Synthase Function. Dig Dis Sci 58, 1507–1515 (2013). https://doi.org/10.1007/s10620-013-2610-4

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