Troglitazone prevents glucose-induced insulin resistance of insulin receptor in rat-1 fibroblasts.
Troglitazone (CS045), a compound belonging to the thiazolidine diones, is being tested as a new oral antidiabetic agent. Evidence exists from animal studies and clinical trials with non-insulin-dependent diabetes mellitus patients that Troglitazone might reduce insulin resistance. The molecular mechanism of this effect is not understood. In this study, we investigated whether Troglitazone might interfere with the mechanism of glucose-induced insulin resistance. Several studies indicate that hyperglycemia reduces the kinase activity of the insulin receptor in different cell types. This effect is paralleled by translocation of several protein kinase C (PKC) isoforms, and it can be prevented by PKC inhibitors, which suggests that glucose-induced receptor desensitization is mediated by activation of PKC. We studied the effect of hyperglycemia on the insulin receptor kinase activity and its modulation by Troglitazone in rat-1 fibroblasts that stably overexpress the human insulin receptor. Before stimulation with insulin (10(-7) M), cells were acutely exposed to hyperglycemic conditions in the absence or presence of Troglitazone (0.01-2 micrograms/ml). The insulin receptor was solubilized from a plasma membrane fraction or whole cell lysates, and proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotted against antiphosphotyrosine and anti-insulin receptor beta-subunit (CT 104) antibodies. Acute hyperglycemia (25 mM glucose) induced a significant inhibition of the insulin receptor kinase (IRK) activity within 30 min (inhibition to 30 +/- 12.5% of maximal insulin-stimulated beta-subunit phosphorylation, n = 9, P < 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)
Kellerer
M
M
Institute for Diabetes Research, Munich, Germany.
Kroder
G
G
Tippmer
S
S
Berti
L
L
Kiehn
R
R
Mosthaf
L
L
Häring
H
H
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000818:Animals; D002460:Cell Line; D002839:Chromans / Q000494:pharmacology*; D005347:Fibroblasts / Q000378:metabolism*; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D007004:Hypoglycemic Agents / Q000494:pharmacology*; D015151:Immunoblotting; D007163:Immunosorbent Techniques; D007333:Insulin Resistance; D007546:Isoquinolines / Q000494:pharmacology; D007700:Kinetics; D010766:Phosphorylation; D019000:Phosphotyrosine; D010879:Piperazines / Q000494:pharmacology; D051381:Rats; D011972:Receptor, Insulin / Q000037:antagonists & inhibitors* / Q000187:drug effects* / Q000378:metabolism; D013844:Thiazoles / Q000494:pharmacology*; D045162:Thiazolidinediones; D000077288:Troglitazone; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism
D002839:Chromans; D007004:Hypoglycemic Agents; D007546:Isoquinolines; D010879:Piperazines; D013844:Thiazoles; D045162:Thiazolidinediones; D019000:Phosphotyrosine; D014443:Tyrosine; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D011972:Receptor, Insulin; D000077288:Troglitazone; D005947:Glucose