Insulin attenuates vascular smooth muscle calcification but increases vascular smooth muscle cell phosphate transport.
Medial artery vascular smooth muscle cell (VSMC) calcification increases the risk of cardiovascular mortality in type 2 diabetes. However, the influence of insulin on VSMC calcification is unclear. We explored the effects of insulin on rat VSMC calcification in vitro and found that in a dose-dependent fashion, insulin attenuates VSMC calcification induced by high phosphate conditions as quantified by the o-cresolphthalein calcium (OCPC) method. In an in vitro model of insulin resistance in which cells are exposed to elevated insulin concentrations and the PI 3-kinase pathway is selectively inhibited, increased VSMC calcification was observed, suggesting that the PI 3-kinase pathway is involved in this attenuating effect of insulin. We postulated that insulin may also have an effect on phosphate or calcium transport in VSMC. We found that insulin increases phosphate transport at 3 and 24 h. This effect was mediated by increased Vmax for phosphate transport but not Km. Because type III sodium-phosphate co-transporters Pit-1 and Pit-2 are found in VSMC, we examined their expression by Western blot and real-time RT-PCR. Insulin stimulates Pit-1 mRNA modestly (*p<0.01 versus control), an effect inhibited by PD98059 but not by wortmannin. Pit-1 protein expression is induced by insulin, an effect also inhibited by PD98059 (*p<0.001 versus insulin alone). Our results suggest a role for insulin in attenuating VSMC calcification which may be disrupted in selective insulin signaling impairment seen in insulin resistance. This effect of insulin contrasts with its effect to induce phosphate transport in VSMC.
Wang
Cecilia C Low
CC
Research Service, Denver Veterans Affairs Medical Center, Denver, CO 80220, United States. Cecilia.Wang@UCHSC.edu
Sorribas
Victor
V
Sharma
Girish
G
Levi
Moshe
M
Draznin
Boris
B
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007328:Insulin / Q000378:metabolism*; D007333:Insulin Resistance; D007700:Kinetics; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D010710:Phosphates / Q000378:metabolism*; D051381:Rats; D012306:Risk; D015398:Signal Transduction; D050608:Sodium-Phosphate Cotransporter Proteins / Q000737:chemistry; D050817:Transcription Factor Pit-1 / Q000378:metabolism
D004791:Enzyme Inhibitors; D007328:Insulin; D010710:Phosphates; D050608:Sodium-Phosphate Cotransporter Proteins; D050817:Transcription Factor Pit-1; D002118:Calcium