Glucocorticoid stimulation of ileal Na+ absorptive cell brush border Na+/H+ exchange and association with an increase in message for NHE-3, an epithelial Na+/H+ exchanger isoform.
Methylprednisolone stimulates rabbit ileal neutral NaCl absorption; and aminoglutethimide, which decreases glucocorticoid levels, decreases NaCl absorption. Studies were carried out to determine the mechanism of these effects and to determine which members of the gene family of mammalian Na+/H+ exchangers were involved. Rabbits were treated subcutaneously with methylprednisolone (40 mg daily for 24 or 72 h), aminoglutethimide (100 mg twice daily for 72 h), or saline as a control. Ileal brush border membranes were prepared by magnesium precipitation, and brush border Na+/H+ exchange was determined by 22Na+ uptake over 3-8 s. The 22Na+ uptake experiments were performed in the presence of a voltage clamp using either valinomycin/potassium or tetramethylammonium/nitrate to eliminate potential contributions by other electrogenic transport processes. Methylprednisolone treatment approximately doubled ileal brush border Na+/H+ exchange, whereas aminoglutethimide led to a 50% decrease in Na+/H+ exchange. These effects were specifically on Na+ uptake with an acid inside pH gradient, whereas diffusive Na+ uptake (no pH gradient), glucose-dependent Na+ uptake, and glucose and Na+ equilibrium volumes were not affected. To determine if the increase in Na+/H+ exchange was associated with an increase in message expression, mRNA levels were measured by ribonuclease protection assay. Methylprednisolone stimulated the NHE-3 mRNA level by 4-6-fold at 24 h, which remained increased at 72 h. In contrast, messages for NHE-1 and NHE-2 were not affected by methylprednisolone. In summary, 1) methylprednisolone stimulation of rabbit ileal Na+ absorption is due to stimulation of ileal villus cell brush border Na+/H+ exchange; 2) basal ileal brush border Na+/H+ exchange is dependent on glucocorticoid levels; and 3) an increase in NHE-3 message, but not in NHE-1 or NHE-2 message, correlates with the stimulation of ileal brush border Na+/H+ exchange. It is likely that NHE-3 is an Na+/H+ exchanger that is involved in ileal Na+ absorption.
Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
C H
CH
Yun
S
S
Gurubhagavatula
S A
SA
Levine
J L
JL
Montgomery
S R
SR
Brant
M E
ME
Cohen
E J
EJ
Cragoe
J
J
Pouyssegur
C M
CM
Tse
M
M
Donowitz
D000616:Aminoglutethimide / Q000494:pharmacology; D000818:Animals; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000235:genetics; D004848:Epithelium / Q000378:metabolism; D005947:Glucose / Q000378:metabolism*; D007082:Ileum / Q000378:metabolism*; D007408:Intestinal Absorption / Q000187:drug effects*; D007413:Intestinal Mucosa / Q000378:metabolism*; D007700:Kinetics; D008297:Male; D008775:Methylprednisolone / Q000494:pharmacology*; D008871:Microvilli / Q000187:drug effects / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D011817:Rabbits; D012016:Reference Values; D012964:Sodium / Q000378:metabolism*; D017923:Sodium-Hydrogen Exchangers; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D013393:Sucrase / Q000378:metabolism
D002352:Carrier Proteins; D012333:RNA, Messenger; D017923:Sodium-Hydrogen Exchangers; D000616:Aminoglutethimide; D012964:Sodium; D013393:Sucrase; D000254:Sodium-Potassium-Exchanging ATPase; D005947:Glucose; D008775:Methylprednisolone