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Cited by 6Ion Transport and Channel RegulationNitric Oxide and Endothelin EffectsGastrointestinal motility and disorders

Direct and specific inhibition of constitutive nitric oxide synthase uniquely regulates brush border membrane Na-absorptive pathways in intestinal epithelial cells

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Palaniappan, Balasubramanian, Sundaram, Uma·2018·Nitric Oxide
DOI 10.1016/j.niox.2018.04.007PMC PMC7237230
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Abstract

Pharmacological manipulations of constitutive nitric oxide (cNO) levels have been shown to have variable effects on Na absorption in vivo and in vitro in different tissues. Species differences, untoward in vivo effects (e.g. ENS, blood flow) and pharmacological non-specificity may account for these confounding observations. Thus, to directly and specifically determine the effect of cNO on brush border membrane Na/H exchange (NHE3) and Na-dependent glucose co-transport (SGLT-1), we inhibited cNO synthase (NOS3) with its siRNA in rat small intestinal epithelial cells (IEC-18) in vitro. As expected, intracellular cNO levels were reduced in siRNA NOS3 transfected cells. In these cells, SGLT-1 was significantly reduced compared to control. In contrast, NHE3 was significantly increased in siRNA NOS3 transfected cells. To determine if SGLT-1 changes were secondary to altered Na/K-ATPase, its activity was measured and found to be increased in NOS3 silenced cells. The mechanism of inhibition of SGLT-1 was secondary to diminished affinity of the co-transporter for glucose in NOS3 silenced cells. In contrast, the mechanism of stimulation of NHE3 is by increasing BBM exchanger numbers in siRNA NOS3 cells while the affinity was unaffected. Western blot studies of immunoreactive BBM proteins also confirmed the kinetic studies. All these data indicates that direct and specific inhibition of NOS3 with its siRNA inhibits SGLT-1 while stimulating NHE3 in the BBM. Thus, cNO uniquely and compensatorily regulates BBM NHE3 and SGLT-1 to maintain cellular Na homeostasis and these unique alterations by cNO are mediated by its intracellular 2nd messenger cGMP.

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  • Intracellular cNO differentially regulates NHE3 and SGLT1 in vitro in IEC-18 cells and in vivo in rabbit small intestine.

    “Furthermore, intracellular cNO was shown to differentially regulate NHE3 and SGLT1 in vitro in IEC-18 cells [23,24,25] and in vivo in rabbit small intestine [25,55,56], which implies functional compensation between the two primary Na absorptive pathways.”

    From Regulation of Enterocyte Brush Border Membrane Primary Na-Absorptive Transporters in Human Intestinal Organoid-Derived Monolayers (2024) · cites this paper as [24]

    The Abstract and Discussion report that reducing intracellular cNO in IEC-18 cells inhibits SGLT-1 and stimulates NHE3. The Introduction reports the same opposing effects after pharmacological NO inhibition in rabbits, but cautions that the in vivo observations may reflect effects in multiple tissues; it does not establish intracellular cNO regulation in rabbit epithelial cells. (LLM, conf=0.86)

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  • The reciprocal regulation between NHE3 and SGLT1 in human enterocytes is consistent with compensatory regulation observed in mammalian intestinal cell models.

    “This is consistent with the compensatory regulation between NHE3 and SGLT1 observed in mammalian intestinal cell models [24,25,27], and therefore serves as proof of concept for using siRNA technology in patient-derived organoid monolayers.”

    From Regulation of Enterocyte Brush Border Membrane Primary Na-Absorptive Transporters in Human Intestinal Organoid-Derived Monolayers (2024) · cites this paper as [24]

  • The differential regulation of NHE3 and SGLT1 by cNO implies functional compensation between the two primary Na absorptive pathways.

    “Furthermore, intracellular cNO was shown to differentially regulate NHE3 and SGLT1 in vitro in IEC-18 cells [23,24,25] and in vivo in rabbit small intestine [25,55,56], which implies functional compensation between the two primary Na absorptive pathways.”

    From Regulation of Enterocyte Brush Border Membrane Primary Na-Absorptive Transporters in Human Intestinal Organoid-Derived Monolayers (2024) · cites this paper as [24]

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Cited by

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Authors
Palaniappan, B., & Sundaram, U.
Affiliations
Marshall University (US)
Corresponding author
Sundaram, Uma
ORCID
Venue
Nitric Oxide
Publisher
Elsevier BV
Published
2018-09
Volume
79
Pages
8-13
ISSN
1089-8603 (ISSN-L)
DOI
10.1016/j.niox.2018.04.007
PMC
PMC7237230
OpenAlex
W2800977872
Cited by
6
References
0
Citations by year
2024: 1 · 2022: 1 · 2021: 1 · 2019: 3
FWCI
0.36
Type
journal article
Language
en
Open access
Licence
Locations
  • Nitric Oxide (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • National Institute of Diabetes and Digestive and Kidney Diseases (DK108054, DK067420)
  • Veteran's Affairs Merit Review (BX003443)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ion Transport and Channel Regulation
  • Health Sciences › Medicine › Physiology › Nitric Oxide and Endothelin Effects
  • Health Sciences › Medicine › Gastroenterology › Gastrointestinal motility and disorders
Keywords
constitutive nitric oxide synthase, brush border membrane, SGLT1, Na+/K+-ATPase, intestinal epithelial cells, small intestine, Na+ homeostasis, cGMP signaling, siRNA knockdown
MeSH terms
Animals, Cells, Cultured, Epithelial Cells, Intestine, Small, Sodium, RNA, Small Interfering, Rats, Nitric Oxide Synthase Type III

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