Advanced glycation end products induce apoptosis and procoagulant activity in cultured human umbilical vein endothelial cells

Abstract

Hyperglycemia and the late products of non-enzymatic glycosylation, called advanced glycation end products (AGEs), play an important role in the development of microvascular complications in diabetes mellitus. Previous studies have reported that a high glucose environment triggered apoptotic changes in human umbilical vein endothelial cells (HUVECs). Therefore, we investigated whether AGEs contribute to the development of apoptosis and prothrombotic activity in HUVECs. After incubation of HUVECs with 0.2, 2.2, 22, 220 and 2200 nM of AGE-bovine serum albumin (BSA) from 6 to 48 h, we assayed the degree of apoptosis and procoagulant activity (PCA). There were no significant differences between HUVECs cultured for 48 h with 0.2, 2.2 or 22 nM of AGE-BSA and in controls in the proportion of apoptotic cells (3.5 +/- 0.8%, 3.9 +/- 1.5% and 5.2 +/- 1.1% vs. 2.5 +/- 0.6%). However, the proportion of apoptotic cells increased significantly to 36.7 +/- 9.8% in 220 nM of AGE-BSA, and 72.3 +/- 10.2% in 2200 nM of AGE-BSA (P < 0.001). PCA levels were 142 +/- 10 s after 6 h of exposure to 22 nM (P < 0.01), 131 +/- 5 s after 6 h of exposure to 220 nM (P < 0.001), and 106 +/- 4 s after 6 h of exposure to 2200 nM of AGE-BSA (P < 0.001). These values show that PCA was shortened significantly from the basal value of 161 +/- 6 s, and remained below the basal level until the end of the study. The amount of tissue factor was also significantly increased in 22 and 220 nM of AGE-BSA compared to the controls. In conclusion, this study showed that AGEs could induce apoptosis and increase procoagulant activity in cultured HUVECs. We suggest that AGEs can contribute to the development of microvascular complications through cell death of HUVECs and functional changes of the blood vessels. (C) 1999 Elsevier Science Ireland Ltd. All rights reserved.