Abstract：
AIMS/HYPOTHESIS:Hyperinsulinaemia and insulin resistance, as well as low IGF-I, have been implicated in the pathogenesis of cardiovascular disease. Little is known about direct effects of IGF-I and insulin on human coronary artery smooth muscle cells (HCASMCs). Our aim was to characterise the expression and function of IGF-I receptor (IGF-IR) and insulin receptor (IR) in HCASMCs. MATERIALS AND METHODS:Cultured HCASMCs were used. mRNA expression was measured by quantitative real-time RT-PCR analysis. Receptor proteins, phosphorylation of beta-subunits and the presence of hybrid IR/IGF-IR were analysed by immunoprecipitation and western blotting. DNA synthesis and glucose metabolism were assessed using [3H]thymidine incorporation and D-[U-14C]glucose accumulation respectively. RESULTS:The mRNA expression of IGF-IR was approximately eight-fold higher than that of IR in HCASMCs. The presence of IGF-IR and IR could be demonstrated by immunoprecipitation and western blot analysis. Phosphorylation of the IGF-IR beta-subunit was obtained by IGF-I at 10(-10)-10(-8) mol/l and insulin at 10(-8) mol/l. Insulin and IGF-I at 10(-10)-10(-9) mol/l phosphorylated the IR beta-subunit. When immunoprecipitated with monoclonal anti-IR alpha-subunit or IGF-IR alpha-subunit antibodies, we found bands in slightly different positions, suggesting the presence of hybrid IR/IGF-IR. IGF-I at 10(-9)-10(-8) mol/l significantly stimulated [3H]thymidine incorporation and at a concentration of 10(-9)-10(-7) mol/l also D-[U-14C]glucose accumulation in HCASMCs. Insulin at 10(-9)-10(-7) mol/l had no effect on DNA synthesis, but increased glucose accumulation at 10(-7) mol/l. CONCLUSIONS/INTERPRETATION:Our study provides experimental evidence that IGF-IR and possibly hybrid IR/IGF-IR play a role in HCASMCs.
journal_name
Diabetologiajournal_title
Diabetologiaauthors
Chisalita SI,Arnqvist HJdoi
10.1007/s00125-005-1890-4subject
Has Abstractpub_date
2005-10-01 00:00:00pages
2155-61issue
10eissn
0012-186Xissn
1432-0428journal_volume
48pub_type
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