High glucose induces apoptosis of human peritoneal mesothelial cells by caspase-3 activation

Objective To investigate the mechanism of high-glucose-induced injury to human peritoneal mesothelial cells(HPMC). Methods (1)The cultured HPMCs were exposed to culture medium containing different concentrations of glucose(1. 5% , 2. 5% , 4. 25% )for 48 hours and 4. 25% mannitol and normal culture medium were as control. Then apoptosis was observed by flow cytometry and caspase-3 activity was measured by ApoAlert?CPP33/Caspase-3 Assay kits. (2) The cultured HPMCs were exposed to 4.25% glucose culture medium containing different concentrations of caspases inhibitor, Z-VAD. fmk (25, 50, 100 μmol/L) for 48 hours and 4. 25% glucose culture medium containing DMSO was as control. Then apoptosis was observed by flow cytometry and caspase-3 activity was measured by ApoAlert?CPP33/ Caspase-3 Assay kits as well. Results (1) Glucose increased caspase-3 activity in a concentration-dependent manner. Compared to control, caspase-3 activity was significantly higher in 4. 25% glucose group and 2. 5% glucose group, but not significantly different in 1. 5% glucose group and 4. 25% mannitol group. (2) Apoptotic rate of HPMC was significantly lower in Z-VAD. fmk group than that in control. Z-VAD. fmk decreased the number of apoptotic cells in a concentration-dependent manner. Also, caspase-3 activity of HPMC was significantly lower in Z-VAD. fmk group than that in control. Conclutions (1) High-glucose can induce apoptosis and caspase-3 activation of HPMC in a dose-dependent manner. (2) Z-VAD. fmk inhibits high glucose-induced apoptosis of HPMC in a dose-dependent manner. (3)High glucose induces apoptosis of human peritoneal mesothelial cells by caspase-3 activation.