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TMP on human umbilical vein endothelial cells (Eahy926) after hypoxia induced by cobalt chloride fibrinolytic function

Author: WuZuo
Tutor: ZhangJinSong;XiaoHang
School: Nanjing Medical University
Course: Emergency Medicine
Keywords: tetramethylpyrazine human umbilical vein endothelial cell line (Eahy926) cobalt chloride (CoCl2) hypoxia fibrinolysis
CLC: R285
Type: Master's thesis
Year: 2010
Downloads: 54
Quote: 0
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Abstract


PartⅠEstablish hypoxia model with human umbilical vein endothelial cell (Eahy926) treated by CoCl2 and different concentration TMP protect Eahy926 cell from hypoxia-induced damage and increase viability of cells.Objective To establish hypoxia model with human umbilical vein endothelial cell line (Eahy926) and discuss the different concentrations of tetramethylpyrazine (TMP) on Eahy926’s injury with hypoxia. Method Different concentrations of CoCl2 induced Eahy926 and established CoCl2 1.6mmol / L induced hypoxia model.The cells were divided into three groups: control group, CoCl2-treated group (1.6 mmol/L) and CoCl2+TMP-treated groups (0.04mg/mL、0.08mg/mL、0.16mg/mL and 0.32mg/mL). Hypoxia was mocked by treatment of CoCl2. Cell viability was evaluated by Hoechst staining and cell counting kit-8 (CCK-8). Results Cell viability was decreased in CoCl2-treated and CoCl2+TMP- groups compared with control group (all P<0.05). However, treatment of TMP improved cell viabilityin a dose-dependent manner (both P < 0.05),except CoCl2+TMP-treated groups (0.32mg/mL). Conclusion TMP can effectively protect Eahy926 cell from hypoxia-induced damage and increase viability of cells. TMP 0.08mg/ml and 0.16mg/ml group in which Eahy926 significantly affected cell viability. [Key words]tetramethylpyrazine; human umbilical vein endothelial cell line (Eahy926); cobalt chloride (CoCl2); hypoxia; Part II Effect of tetramethylpyrazine on fibrinolytic function in human umbilical vein endothelial cell (Eahy926) treated by CoCl2Abstract: Objective: To establish hypoxia model with human umbilical vein endothelial cell line (Eahy926) and investigate the effect of tetramethylpyrazine (TMP) on Eahy926’s fibrinolytic function with hypoxia. Methods: The cells were divided into three groups: control group, CoCl2-treated group (1.6 mmol/L) and CoCl2+TMP-treated groups (0.08 and 0.16mg/mL). Hypoxia was mocked by treatment of CoCl2. Cell viability was evaluated by Hoechst staining and cell counting kit-8 (CCK-8); the expression of t-PA、u-PA and PAI-1 was measured by semi-quantitative RT-PCR; t-PA, u-PA and PAI-1 in the supernatant of culture medium were detected by ELISA. Results: Cell viability was decreased in CoCl2-treated and CoCl2+TMP- groups compared with control group (P<0.01). However, treatment of TMP improved cell viabilityin a dose-dependent manner (P<0.01). Moreover, CoCl2 induced a decreasing mRNA expression (P<0.01) and secretion of t-PA (P<0.01); TMP treatment elevated t-PA mRNA expression and contents of the cell supernatant (P<0.01) in a dose-dependent way. Conclusions: TMP can effectively protect Eahy926 cell from hypoxia-induced damage and increase viability of cells in a concentration-dependent manner; TMP might enhance the fibrinolytic function of endothelial cells by regulating the expression and secretion of t-PA.

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