Chronic estrogen deficiency causes gastroparesis by altering neuronal nitric oxide synthase function

K. Ravella, Ayman Al-Hendy, C. Sharan, A. B. Hale, K. M. Channon, S. Srinivasan, P. R. Gangula

Research output: Contribution to journalArticle

14 Citations (Scopus)

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 BH 4 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.

Original languageEnglish (US)
Pages (from-to)1507-1515
Number of pages9
JournalDigestive Diseases and Sciences
Volume58
Issue number6
DOIs
StatePublished - Jun 1 2013

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Gastroparesis
Nitric Oxide Synthase Type I
Estrogens
Biopterin
Pyloric Antrum
Stomach
Gastric Emptying
Dimerization
FSH Receptors
Sexism
Knockout Mice
Estrogen Receptors
Electric Stimulation
Area Under Curve
Breeding
Estradiol
Fasting
Down-Regulation
Western Blotting
High Pressure Liquid Chromatography

Keywords

  • Biopterins
  • Estrogen
  • Follicle stimulating hormone
  • Gastroparesis
  • Nitric oxide
  • nNOS dimerization

ASJC Scopus subject areas

  • Gastroenterology
  • Physiology

Cite this

Ravella, K., Al-Hendy, A., Sharan, C., Hale, A. B., Channon, K. M., Srinivasan, S., & Gangula, P. R. (2013). Chronic estrogen deficiency causes gastroparesis by altering neuronal nitric oxide synthase function. Digestive Diseases and Sciences, 58(6), 1507-1515. https://doi.org/10.1007/s10620-013-2610-4

Chronic estrogen deficiency causes gastroparesis by altering neuronal nitric oxide synthase function. / Ravella, K.; Al-Hendy, Ayman; Sharan, C.; Hale, A. B.; Channon, K. M.; Srinivasan, S.; Gangula, P. R.

In: Digestive Diseases and Sciences, Vol. 58, No. 6, 01.06.2013, p. 1507-1515.

Research output: Contribution to journalArticle

Ravella, K, Al-Hendy, A, Sharan, C, Hale, AB, Channon, KM, Srinivasan, S & Gangula, PR 2013, 'Chronic estrogen deficiency causes gastroparesis by altering neuronal nitric oxide synthase function', Digestive Diseases and Sciences, vol. 58, no. 6, pp. 1507-1515. https://doi.org/10.1007/s10620-013-2610-4
Ravella, K. ; Al-Hendy, Ayman ; Sharan, C. ; Hale, A. B. ; Channon, K. M. ; Srinivasan, S. ; Gangula, P. R. / Chronic estrogen deficiency causes gastroparesis by altering neuronal nitric oxide synthase function. In: Digestive Diseases and Sciences. 2013 ; Vol. 58, No. 6. pp. 1507-1515.
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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 BH 4 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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AU - Ravella, K.

AU - Al-Hendy, Ayman

AU - Sharan, C.

AU - Hale, A. B.

AU - Channon, K. M.

AU - Srinivasan, S.

AU - Gangula, P. R.

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N2 - 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 BH 4 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.

AB - 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 BH 4 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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KW - Estrogen

KW - Follicle stimulating hormone

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KW - Nitric oxide

KW - nNOS dimerization

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