Toxicity Features of High Glucose on Endothelial Cell Cycle and Protection by Dan Gua-Fang(丹瓜方) in ECV-304 in High Glucose Medium

Objective： To study the toxicity features of high glucose on the endothelial cell cycle and the influence of Dan Gua-Fang , a Chinese herbal compound prescription, on the reproductive cycle of vascular endothelial cells cultivated under a high glucose condition; to reveal the partial mechanisms of Dan Gua- Fang in the prevention and treatment of endothelial injury caused by hyperglycemia in diabetes mellitus （DM）; and offer a reference for dealing with the vascular complications of DM patients with long-term high blood glucose. Methods： Based on the previous 3-（4,5）-dimethylthiahiazo （z-yl）-3-5-diphenytetrazoliumromide （MI-I-） experiment, under different medium concentrations of glucose and Dangua liquor, the endothelial cells of vein-304 （ECV-304） were divided into 6 groups as follows： standard culture group （Group A, 5.56 mmol/L glucose）; 11300 herb-standard group （Group B）; high glucose culture group （Group C, 16.67 mmol/L glucose）; 1/150 herb-high glucose group （Group D）; 1/300 herb-high glucose group （Group E）; and 1/600 herb-high glucose group （Group F）. The cell cycle was assayed using flow cytometry after cells were cultivated for 36, 72 and 108 h, respectively. Results： （1） The percentage of cells in the Go/G1 phase was significantly increased in Group C compared with that in Group A （P〈0.05）, while the percentage of S-phase （S%） cells in Group C was significantly reduced compared with Group A （P〈0.05）; the latter difference was dynamically related to the length of growing time of the endothelial cells in a high glucose environment. （2） The S% cells in Group A was decreased by 30.25% （from 40.23% to 28.06%） from 36 h to 72 h, and 12.33% （from 28.06% to 24.60%） from 72 h to 108 h; while in Group C, the corresponding decreases were 23.05% and 21.87%, respectively. The difference of S% cells between the two groups reached statistical significance at 108 h （P〈0.05）, （3） The percentage difference of cells in the Gz/M phase between Group C and Group A was statistically significant at 72 h （P〈0.01）. （4） 1/300 Dan Gua-Fang completely reversed the harmful effect caused by 16.67 mmol/L high glucose on the cell cycle; moreover it did not disturb the cell cycle when the cell was cultivated in a glucose concentration of 5.56 mmol/L. Conclusions： High glucose produces an independent impact on the cell cycle. Persistent blocking of the cell cycle and its arrest at the Go/G1 phase are toxic effects of high glucose on the endothelial cell cycle. The corresponding variation of the arrest appears in the S phase. 1/300 Dan Gua-Fang completely eliminates the blockage of high glucose on the endothelial cell cycle.