Adult rat cardiomyocytes were maintained in normal (NG, 5.5 mM) or high glucose (HG, 25.5 mM) medium for 12 h. Contractile and intracellular Ca2 + properties were measured using a video edge-detection system including peak shortening (PS), maximal velocity of shortening/relengthening (¡À dL/dt), time-to-PS (TPS), time-to-90 % relengthening (TR90), rise in intracellular Ca2 + Fura-2 fluorescence intensity and intracellular Ca2 + decay. Production of ROS/O2? and mitochondrial integrity were examined using fluorescence imaging, aconitase activity and Western blotting.
High glucose triggered abnormal contractile and intracellular Ca2 + properties including reduced PS, ¡À dL/dt, prolonged TR90, decreased electrically-stimulated rise in intracellular Ca2 + and delayed intracellular Ca2 + clearance, the effects of which were ablated by the PKC¦ÂII inhibitor LY333531. Inhibition of PKC¦ÂII rescued glucose toxicity-induced generation of ROS and O2?, apoptosis, cell death and mitochondrial injury (reduced aconitase activity, UCP-2 and PGC-1¦Á). In vitro studies revealed that PKC¦ÂII inhibition-induced beneficial effects were mimicked by the NADPH oxidase inhibitor apocynin and were canceled off by mitochondrial uncoupling using FCCP.
These findings suggest the therapeutic potential of specific inhibition of PKC¦ÂII isoform in the management of hyperglycemia-induced cardiac complications.