Antisense GLUT-1 protects mesangial cells from glucose induction of GLUT-1 and fibronectin expression.
A stable clone of rat mesangial cells expressing antisense GLUT-1 (i.e., MCGT1AS cells) was developed to protect them from high glucose exposure. GLUT-1 protein was reduced 50%, and the 2-deoxy-[(3)H]glucose uptake rate was reduced 33% in MCGT1AS. MCLacZ control cells and MCGT1 GLUT-1-overexpressing cells were used for comparisons. In MCLacZ, 20 mM D-glucose increased GLUT-1 transcription 90% vs. no increase in MCGT1AS. Glucose (8 mM) and 12 mM xylitol [a hexose monophosphate (HMP) shunt substrate] did not stimulate GLUT-1 transcription. An 87% replacement of the standard 8 mM D-glucose with 3-O-methylglucose reduced GLUT-1 transcription 80%. D-Glucose (20 mM) increased fibronectin mRNA and protein by 47 and 100%, respectively, in MCLacZ vs. no increases in MCGT1AS. Fibronectin synthesis was elevated 48% in MCGT1 and reduced 44% in MCGT1AS. We conclude that 1) transcription of GLUT-1 in response to D-glucose depends on glucose metabolism, although not through the HMP shunt, and 2) antisense GLUT-1 treatment of mesangial cells blocks D-glucose-induced GLUT-1 and fibronectin expression, thereby demonstrating a protective effect that could be beneficial in the setting of diabetes.
Heilig
C W
CW
Division of Nephrology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA. cheilig@welch.jhu.edu
Kreisberg
J I
JI
Freytag
S
S
Murakami
T
T
Ebina
Y
Y
Guo
L
L
Heilig
K
K
Loberg
R
R
Qu
X
X
Jin
Y
Y
Henry
D
D
Brosius
F C
FC
D018528:ATP-Binding Cassette Transporters / Q000235:genetics* / Q000378:metabolism; D027322:Amino Acid Transport System X-AG; D000818:Animals; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D002999:Clone Cells; D016373:DNA, Antisense / Q000494:pharmacology; D005353:Fibronectins / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D005920:Glomerular Mesangium / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D005947:Glucose / Q000494:pharmacology*; D007763:Lac Operon; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D014161:Transduction, Genetic
D018528:ATP-Binding Cassette Transporters; D027322:Amino Acid Transport System X-AG; D016373:DNA, Antisense; D005353:Fibronectins; D012333:RNA, Messenger; D015500:Chloramphenicol O-Acetyltransferase; D005947:Glucose