The role of the imino transporter protein in sepsis-impaired intestinal proline absorption.
Recently, sepsis has been shown to impair intestinal amino acid absorption in addition to gut metabolic and barrier functions. We investigated intestinal proline absorption in a rabbit model of sepsis. Twelve hours after intraperitoneal injection of lipopolysaccharide, proline uptake by everted jejunal sacs prepared from septic animals (480.4 +/- 67.4 nmol per sac per hour) was significantly reduced compared with controls (846.8 +/- 73.5 nmol per sac per hour) (p < .001 by t test). We next investigated whether reduced expression of transporter proteins contributed to the impaired intestinal proline uptake during sepsis. The proline (imino) carrier of rabbit jejunum is covalently bound by fluorescein isothiocyanate (FITC) and/or phenylisothiocyanate with irreversible inhibition of proline uptake. This binding and inhibition is prevented by sodium chloride and L-proline. Single-cell suspensions of rabbit enterocytes were prepared 12 hours after intraperitoneal injection of lipopolysaccharide/saline or saline alone. Enterocytes were incubated for 30 minutes in tris(hydroxymethyl)aminomethane/ethylenediaminetetraacetate (Tris/EDTA) buffer; buffer with 1 mM phenylisothiocyanate; or buffer with 10 mM proline, 100 mM sodium chloride, and 1 mM phenylisothiocyanate. After incubation with 10 microM FITC in Tris/EDTA buffer for 15 minutes, the percent positivity and fluorescent intensity of FITC binding to enterocytes were determined by using flow cytometry. Sepsis significantly reduced the percentage of enterocytes binding FITC and the fluorescent intensity of FITC binding of proline/sodium chloride-pretreated or untreated cells. This suggests that sepsis depresses the expression of imino transporters by rabbit enterocytes, which may explain the reduced intestinal proline absorption.
Gardiner
K
K
Department of Surgery, Sinai Hospital, Baltimore, MD 21215.
Barbul
A
A
D026921:Amino Acid Transport Systems, Neutral; D000818:Animals; D016470:Bacteremia / Q000378:metabolism*; D002352:Carrier Proteins / Q000096:biosynthesis / Q000502:physiology*; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D004731:Endotoxins / Q000097:blood; D004912:Erythrocytes / Q000378:metabolism; D004926:Escherichia coli; D004927:Escherichia coli Infections / Q000378:metabolism*; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate / Q000494:pharmacology; D066298:In Vitro Techniques; D007408:Intestinal Absorption / Q000187:drug effects / Q000502:physiology*; D017879:Isothiocyanates; D007583:Jejunum / Q000378:metabolism*; D008297:Male; D011392:Proline / Q000378:metabolism*; D011817:Rabbits; D012965:Sodium Chloride / Q000494:pharmacology; D013393:Sucrase / Q000096:biosynthesis; D027981:Symporters; D013861:Thiocyanates / Q000378:metabolism
D026921:Amino Acid Transport Systems, Neutral; D002352:Carrier Proteins; D004731:Endotoxins; D017879:Isothiocyanates; D027981:Symporters; D013861:Thiocyanates; C041832:proline transport protein; C005441:phenylisothiocyanate; D012965:Sodium Chloride; C044101:endotoxin, Escherichia coli; D011392:Proline; D013393:Sucrase; D016650:Fluorescein-5-isothiocyanate