Protectice Effect Of Scallop Skirt-Glycosaminoglycan On Vascular Endothelial Cell Injured By Hydrogen Peroxide

Objectives: To investigate the protective action of scallop skirt-glycosaminoglycan (SS-GAG) on the vascular endothelial cell, and to study its mechanism of anti-atherosclerosis (AS)Methods: 1,The endothelial cell strain of human umbilical. vein (HUVEC, ECV304) had been cultured in vitro, and we established a modal of endothelial cell oxidative damage induced by hydrogen peroxide (H₂O₂)2,In vitro it took a model of endothelial cell oxidative damage induced by H₂O₂. We tested the influence of SS-GAG on the proliferation activity of endothelial cell oxidative damage by the means of MTT assay. 3,We used chemical methods to analysis the content of Lactate Dehydrogenase (LDH) in order to study the effect of SS-GAG on endothelial cell oxidative damage induced by H₂O₂. 4,In order to observe the effect of SS-GAG on endothelial cell oxidative damage induced by H₂O₂, chemical methods was used to analysis the activity of Glutathione Peroxidase (GSH-PX) and the level of the Total Antioxidative Capacity (T-AOC). 5,We used chemical methods to analysis the content of nitric oxide (NO) and the activity of endothelial nitric oxide synthase (NOS) in order to study the effect of SS-GAG on endothelial cell oxidative damage induced by hydrogen peroxide (H₂O₂). 6,In order to observe the effect of SS-GAG on endothelial cell oxidative damage induced by H₂O₂, xanthne oxides method was used to analysis the activity of super oxide dismutase(SOD) and the level of malanyldiadeyhde(MDA). 7,By the means of radio-immunity, we observed the action of SS-GAG on Prostacyclin- F_1α(PGF_1α) and Thromboxane-B₂(TXB₂) contents of the endothelial cells oxidative injured by H₂O₂. 8,We used radio-immunity methods to analysis the content of Endothelin (ET) in order·to study the effect of SS-GAG on endothelial cell oxidative damage induced by H₂O₂.Results: 1,In model groups damaged by H₂O₂, some certain of injuries were observed. While the groups damaged by H₂O₂(terminal concentration were 75,150,300,600,1500,3000,6000μmol/L), the endothelial cells showed obvious abnormalities. 2,In model groups damaged by H₂O₂, when compared with normal control groups, the proliferation activity of endothelial cells remarkably decreased (p＜0. 01); In high dose model groups damaged by H₂O₂, the proliferation activity were lower than those in low dose model groups (p＜0. 05). 3,LDH testing result showed that the LDH activity of model group was much higher than that in normal control group (p＜0. 01); While the groups pretreated by SS-GAG(terminal concentration were 50μg/ml, 100μg/ml, 200μg/ml), compared with model group the LDH activity obsiously decreased (p＜0. 01). 4,Compared with normal control groups, in model groups damaged by H₂O₂, the level GSH-PX and T-AOC of endothelial cells decreased remarkably (p＜0. 01), but it increased obviously when pretreated by SS-GAG(terminal concentration were 50μg/ml, 100μg/ml, 200μg/ml) (p＜0. 01). 5,Compared with normal control group, the level of NO and the activity of NOS testing results showed that NO level and NOS activity of positive control group were lower (p＜0. 01); While the groups pretreated by SS-GAG(terminal concentration were 50μg/ml, 100μg/ml, 200μg/ml) the level of NO and the activity of NOS obviously increasde (p＜0. 01). 6,In order to observe the effect of SS-GAG on endothelial cell oxidative damage induced by H₂O₂, Xanthine Oxidase method was used to analysis the activity of superoxide dismutase(SOD) and TBARS method was used to analysis the level of malanyldiadeyhde (MDA) within cells. 7,PGF_1α and TX B₂ testing results showed that PGF_1α level of model groups much lower than normal control group (p＜0. 01); While the groups pretreated by SS-GAG (terminal concentration were 50μg/ml, 100μg/ml, 200μg/ml), compared with model groups the PGF_1α level obviously increasde (p＜0. 01). TX B₂ testing results showed that TX B₂ level of model groups much higher than normal control group (p＜0. 01); While the groups pretreated by SS-GAG (terminal concentration were 50μg/ml, 100μg/ml, 200μg/ml), compared with model groups the TX B₂ level obviously decreasde (p＜0. 01). 8,Compared with normal control group, the level of ET testing results showed that ET activity of positive control group were higher (p＜0. 01); While the groups pretreated by SS-GAG(terminal concentration were 50μg/ml, 100μg/ml, 200μg/ml) the level of ET obviously decreasde (p＜0. 01).Conclusion: SS-GAG can lessen proliferatory inhibition of endothelial cell induced by H₂O₂, decrease the level of MDA and LDH, promote the activity of SOD, increase the excretion of NO and NOS, enhance the level of PGF_1α, and decrease the level of ET and TXB₂According to these studies, they can indicate that SS-GAG has the protective acion on oxidative damage of endothelial cell induced by H₂O₂. This study provided new theoretical basis to the prevention and treatment of AS.