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 ce...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.展开更多
基金Supported by the Post-doctoral Science Funds of China(No. 20070410128)CHEN Ke-ji Development Fund of Integrative Medicine(No.CKJ2006001)Science Fund of Health Department of Fujian Province,China(No.WZY0618)
文摘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.