Enhanced expression of group IIA secreted phospholipase A2 by elevated glucose levels in cytokine-stimulated rat mesangial cells and in kidneys of diabetic rats.
BACKGROUND
Group IIA secreted phospholipase A2 (sPLA2-IIA) has been implicated in various inflammatory processes including the kidney. Previously it has been shown that potent proinflammatory cytokines such as interleukin 1beta (IL-1beta) increase sPLA2-IIA expression and secretion in rat mesangial cells.


OBJECTIVE
The present study examines the effects of glucose on sPLA2-IIA regulation in interleukin-1beta (IL-1beta) treated mesangial cell cultures and in diabetic kidneys of Sprague-Dawley rats.


METHODS
Rat mesangial cells were grown either in low glucose (5.55 mM D-Glucose) or high glucose (25 mM) conditions followed by assessment of sPLA2-IIA transcription, expression and secretion after 24 h. The model of streptozotocin induced diabetes mellitus in Sprague-Dawley rats was used for the in vivo experiments. Diabetic kidneys where examined for sPLA2-IIA-mRNA and -protein expression as well as IL-1beta-levels at 2, 4 and 6 weeks after induction of diabetes mellitus.


RESULTS
Increased concentration of glucose had a weak, but significant stimulatory effect on sPLA2-IIA expression, which was markedly up-regulated (2-3-fold) in IL-1beta treated mesangial cells compared to the levels obtained in low glucose medium. Concerning the underlying mechanism, we found that high concentration of glucose increased the activity of the rat sPLA2-IIA-promoter, whereas mRNA stability was not affected. Furthermore, the in vivo experiments revealed a marked up-regulation of sPLA2-IIA mRNA and protein in the diabetic rat kidneys 2 - 4 weeks after induction. Since the strong up-regulation of sPLA2-IIA in vitro under high glucose conditions occurred mainly in presence of cytokine, we measured the levels of IL-1beta in diabetic kidneys by ELISA. We detected rat IL-1beta only in diabetic, but not in control rat kidneys.


CONCLUSIONS
The changes of sPLA2-IIA expression under increased glucose concentrations as well as in diabetic rat kidneys suggest a function of this enzyme as an acute phase protein providing lipid autacoids that may contribute to early changes in the course of diabetic nephropathy.
Vlachojannis
G J
GJ
Pharmazentrum Frankfurt, Germany.
Scholz-Pedretti
K
K
Fierlbeck
W
W
Geiger
H
H
Pfeilschifter
J
J
Kaszkin
M
M
D000704:Analysis of Variance; D000818:Animals; D015152:Blotting, Northern; D002478:Cells, Cultured; D016207:Cytokines / Q000494:pharmacology*; D003921:Diabetes Mellitus, Experimental / Q000503:physiopathology; D003928:Diabetic Nephropathies / Q000201:enzymology* / Q000503:physiopathology; D004195:Disease Models, Animal; D004790:Enzyme Induction; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005786:Gene Expression Regulation; D005920:Glomerular Mesangium / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology*; D006943:Hyperglycemia / Q000175:diagnosis / Q000201:enzymology*; D007668:Kidney / Q000378:metabolism / Q000503:physiopathology; D008297:Male; D010741:Phospholipases A / Q000235:genetics / Q000378:metabolism*; D054467:Phospholipases A2; D011336:Probability; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D012680:Sensitivity and Specificity; D013311:Streptozocin
D016207:Cytokines; D012333:RNA, Messenger; D013311:Streptozocin; D010741:Phospholipases A; D054467:Phospholipases A2