Effect of high glucose concentration on expression of coagulation and anticoagulation factors in Hep G2 cells

• Main Authors: Yi-Chien Lai, 賴儀倩
• Other Authors: Kung-Chi Chan
• Format: Others
• Language: zh-TW
• Published: 2006
• Online Access: http://ndltd.ncl.edu.tw/handle/pa26x9

碩士 === 靜宜大學 === 食品營養研究所 === 94 === More than 80% diabetic patients die from a thrombotic cardiovascular event, and hyperglycemia-induced oxidative damage may play an important role in the pathogenesis of thrombotic diseases. Studies from clinical trials reported that diabetic patients have abnormal activity of blood coagulation, however, the mechanism of how hyperglycemia affects coagulation cascade is still unclear. Therefore, the purpose of this study was to investigate the effect of high glucose concentration on the expression of coagulation and anticoagulation factor in HepG2 cells. In the 1st study, high glucose concentration was first determined for the subsequent experiment. HepG2 cells were incubated under different glucose (25~65mM) and osmotic (619~659 mOsm/L) concentrations in the medium for 4, 6, 8, 10 and 12 consecutive days, and cells survival rate was measured by the MTT assay. Malondialdehyde (MDA) concentration also was measured by TBA assay at 10th day of incubation. The results of the 1st study showed that HepG2 cells incubated at 45mM glucose concentration have similar growth curve with cells incubated under normal glucose concentration(25mM). However, MDA production was significantly increased in cells with high glucose concentration. In the 2nd experiment, HepG2 cells were incubated under 25mM glucose (control) and 45mM glucose in DMEM medium. After 10 days of incubation, cell survival and protein kinase C (PKC) activity of cell lysate, total protein, content of incubating medium, MDA, C-reactive protein (CRP), activity and concentration of Factor VII and antithrombin III (AT-III) were measured. The results of 2nd study showed that MDA production was induced significantly by high glucose (45mM) concentration. Activities of PKC and Factor VII were increased, and AT-III activity was decreased (p<0.05) under high glucose treatment. High glucose concentration inhibited Factor VII expression, but had no effect on AT-III secretion in HepG2 cells. In conclusion, hyperglycemia-induced lipid peroxidation and altered PKC activity may involved in the posttranslational regulation of abnormal activity of factor VII and AT-III.