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A Role of Epigallocatechin-3-gallate on Oxidative Stress in Endothelial Cells

Author: HanZuo
Tutor: LiuJing
School: Lanzhou University
Course: Internal Medicine
Keywords: EGCG human umbilical vein endothelial cells oxidative stress apoptosis NF-kappa B high glucose
CLC: R285
Type: Master's thesis
Year: 2009
Downloads: 19
Quote: 0
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Abstract


Objective To research the effect of differnt concentration Epigallocatechin-3-gallate(EGCG) on the human umbilical vein endothelial cells(HUVEC) in vitro by high glucose which may induce the apoptosis of HUVEC cells via increasing intracellular oxidative stress formation, in which the ratio of human nuclear factor P65(NF-κB p65) expression seems to be involved.Methods HUVEC were isolated from freshly collected umbilical cords. High glucose which induced the oxidative stress that caused the ratio of NF-κB P65 expression on HUVEC cells. Experiment samples were co-cultured with EGCG at different concentration (12.5, 25, 50, 100, 200κmol/L). The effect of immunocytochemistry staining on the NF-κB P65 expression in HUVEC, and The rate of NF-κB P65 mRNA was analyzed by RT-PCR . The rate of apoptoic cells was analyzed by AV-PI stained flow cytometry(FCM).Results High glucose could induce the oxidative stress of HUVEC and the apoptosis of HUVEC that high glucose group from 12h 15.14±0.32 % to 48h 33.38±1.01 %, (P<0.05) compared with normal control group; up-regulation of the ratio of NF-κB P65 mRNA expression, from 12h 0.46±0.02 to 48h 0.72±0.03 (P<0.05) comparsed with high glucose group; some concentration of EGCG (50-200μmol/L) that could decrease the ratio of NF-κB P65 mRNA expression and the decrease had some relation with the increased of the concentration of EGCG, there is a dependent relationship of dose-effect to time-effect. The same time EGCG can down-regulation of the ratio of cellular apoptosis, all group could have some protection to the cellular apoptosis (P<0.05) , except 12h EGCG 12.5μmol/L. They had a positive correlation relationship.Conclusions The expression of NF-κB P65 mRNA will decrease with theenlargement concentration of EGCG. the expression of NF-κB P65 mRNA may playan important role in the survival/ apoptosis of HUVEC induced by high glucose.EGCG could significantly protect HUVEC though inhibiting the up-regulation ofNF-κB signal transduction pathway induced by high glucose, and could significantlydecrease apoptosis through this pathway.

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