Hyperglycaemia, nestin expression and DNA synthesis in VSMCs. (Panels A & B) Exposure of 1st/2nd passage carotid artery VSMCs to 30 mM mannitol or 30 mM L-glucose for 24 hours did not influence nestin protein expression as compared to DMEM. A 24 and/or 48-hour exposure to 30 mM D-glucose significantly reduced nestin protein levels, whereas smooth muscle α-actin (SMA), caldesmon and smooth muscle-22α protein expression was unchanged. (*) Denotes P < 0.05 versus L-glucose and protein levels normalized to GAPDH. (Panel C) VSMCs exposed to 30 mM D-glucose for 24 hours attenuated 3H-Thymidine uptake. VSMCs treated with 10% FBS increased 3H-thymidine uptake, whereas 30 mM mannitol and 30 mM L-glucose had no effect (Panel D) VSMCs exposed to 30 mM D-glucose, 30 mM L-glucose or 30 mM mannitol for 24 hours had no effect on 3H-Leucine uptake, whereas a robust increase was observed with 10% FBS. Results are presented as % change versus DMEM and (*) denotes P < 0.05 versus L-glucose/mannitol.