LPS/CD14 activation triggers SGLT-1-mediated glucose uptake and cell rescue in intestinal epithelial cells via early apoptotic signals upstream of caspase-3.
Recent findings indicate that enhanced glucose uptake protects enterocytes from excessive apoptosis and barrier defects induced by LPS exposure. The aim of this study was to characterize the mechanisms responsible for increased sodium-dependent glucose cotransporter (SGLT)-1 activity in enterocytes challenged with LPS. SGLT-1-transfected Caco-2 cells were incubated with LPS in high glucose media. LPS increased SGLT-1 activity in dose- and time-dependent fashion, and is due to increased V(max) of the cotransporter. Elevated apical expression of SGLT-1 was also demonstrated. This LPS-induced effect was colchicine-inhibitable, suggesting microtubule-dependent translocation of SGLT-1 onto apical surface. Immunofluorescence staining showed expression of CD14 on the apical surface, but no TLR-4, on these cells. Neutralizing anti-CD14 decreased the LPS-induced upregulation of SGLT-1 activity, whereas anti-TLR-4 had no effect. Pharmacological studies indicated that signaling for LPS-mediated SGLT-1 glucose uptake depends on caspase-8 and -9 activation, but occurs independently of caspase-3. The findings describe a novel feedback mechanism within the apoptotic signaling pathway for SGLT-1-dependent cytoprotection. The observation suggests a new function for CD14 on enterocytes, involving the induction of the caspase-dependent SGLT-1 activity, which ultimately leads to cell rescue. The understanding of these signaling events may shed light on enterocytic cytoprotection and homeostasis mechanism upon pro-apoptotic challenges.
Yu
Linda C H
LC
Department of Biological Sciences, BI 117 Mucosal Inflammation Research Group, University of Calgary, 2500 University Dr. N.W., Calgary, AB, T2N 1N4, Canada.
Turner
Jerrold R
JR
Buret
Andre G
AG
D017209:Apoptosis; D018938:Caco-2 Cells; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism*; D019610:Cytoprotection / Q000502:physiology; D020895:Enterocytes / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D018950:Lipopolysaccharide Receptors / Q000378:metabolism*; D008070:Lipopolysaccharides / Q000378:metabolism*; D015398:Signal Transduction; D051269:Sodium-Glucose Transporter 1 / Q000378:metabolism*; D014162:Transfection
D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; C495674:SLC5A1 protein, human; D051269:Sodium-Glucose Transporter 1; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; D005947:Glucose