Toxicity features of high glucose on endothelial cell cycle and protection by Dan Gua-Fang in ECV-304 in high glucose medium / 中国结合医学杂志

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>Based on the previous 3-(4,5)-dimethylthiahiazo (z-y1)-3-5-diphenytetrazoliumromide (MTT) 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); 1/300 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.</p><p><b>RESULTS</b>(1) The percentage of cells in the G0/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 G2/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.</p><p><b>CONCLUSIONS</b>High glucose produces an independent impact on the cell cycle. Persistent blocking of the cell cycle and its arrest at the G0/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.</p>

Anticolchicine cytotoxicity enhanced by Dan Gua-Fang, a Chinese herb prescription in ECV304 in mediums / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the effect of anticolchicine cytotoxicity of Dan Gua-Fang, a Chinesea Chinese), a Chinese herbal compound prescription on endothelial cells of vein (ECV304) cultivated in mediums of different glucose concentrations as well as the proliferation of those cells in the same conditions, in order to reveal the value of Dan Gua-Fang in preventing and treating endothelial damage caused by hyperglycemia in diabetes mellitus.</p><p><b>METHODS</b>The research was designed as three stages. The growing state and morphological changes were observed when ECV304 were cultivated in the culture mediums, which have different glucose concentrations with or without Dan Gua-Fang and at the same time with or without colchicine.</p><p><b>RESULTS</b>(1) Dan Gua-Fang at all concentrations reduced the floating cell population of ECV304 cultivated in hyperglycemia mediums. (2) Dan Gua-Fang at all concentrations and hyperglycemia both had a function of promoting "pseudopod-like" structure formation in cultivated ECV304, but the function was not superimposed in mediums containing both hyperglycemia and Dan Gua-Fang. (3) Colchicine reduced and even vanished the "pseudopod-like" structure of the endotheliocyte apparently cultivated in mediums of hyperglycemia or with Dan Gua-Fang. The "pseudopod-like" structure of the endotheliocyte emerged quickly in Dan Gua-Fang groups after colchicine was removed, but it was not the case in hyperglycemia only without Dan Gua-Fang groups. (4) Dan Gua-Fang reduced the mortality of cells cultivated in mediums containing colchicine. The cell revived to its normal state fast after colchicine was removed.</p><p><b>CONCLUSION</b>Dan Gua-Fang has the functions of promoting the formation of cytoskeleton and fighting against colchicine cytotoxicity.</p>

Glucose endothelial cytotoxicity and protection by Dan Gua-Fang, a Chinese herb prescription in huVEC in hyperglycemia medium.

BACKGROUND: Low success rate of blood glucose in diabetes is an international problem. The endothelia cytotoxicity of hyperglycemia has been widely accepted. However, it has not been seen in reports of the value of concentration of high glucose beginning to produce cytotoxicity and the relationship between hyperglycemia and cytotoxicity as well as how to effectively prevent and control hyperglycemia cytotoxicity. Dan Gua prescription is an effective Chinese herb prescription for diabetic vascular complications. METHODS: Dan Gua prescription was contained in Dan Gua liquor utilized in experiments. (1) The cytotoxicity experiment of Dan Gua was carried out with M199 medium whose glucose (Glu) was 5.55 mmol/l to seek for a suitable experimental concentration of Dan Gua. (2) The human vessel endotheliocyte was cultivated for 72 h with mediums containing glucose in different concentrations (Group G1 to Group G11, Glu: 5.5 to 99.9 mmol/l, respectively), and assayed an optical density (OD) value using the 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide method. (3) Experiment 2 was repeated. However, the medium of each group (Groups Y1 to Y11) contained Dan Gua liquor whose concentration was 1/300. RESULTS: There was a negative correlation between means of cell OD values and glucose concentrations (r=-.927, R(2)=.844), and it presented a notable linear correlation (y=0.681-0.002x). Based on the OD value of 5.5-mmol/l glucose concentration (group G1), when glucose concentration reached 22.2 mmol/l (G4), the difference in OD values has a statistical significance. OD values in Y1-Y11 were not less than that of G1. CONCLUSION: There is a notable linear correlation between the endothelial cytotoxicities of Glu and its concentrations. The spinodal point concentration of statistical significance of hyperglycemia cytotoxicity is 22.2 mmol/l; 1/300 Dan Gua can reverse the endothelia cytotoxicity in different concentrations of hyperglycemia.