Experimental Study On The Protective Effect And Mechanism Of Scallop Skirt-glycosaminoglycan On Vascular Endothelial Cell Injury

Objectives: To investigation the protective action of scallop skirt-glycosaminoglycan (SS- GAG) on the endothelial cell, and to study its mechanism of anti-atherosclerosis (AS).Methods: 1. The endothelial cell strain of human umbilical vein (HUVEC,CRL-2480) had been cultured in vitro, and we established a model of endothelial cell oxidative damage induced by oxidized low density lipoprotein (OX-LDL). MTT assay was used to test the influence of SS-GAG on the proliferation activity of endothelial cell oxidative damaged by low density lipoprotein (OX-LDL).2. In vitro it took a model of endothelial cell oxidative damage induced by hydrogen peroxide (H2O2). we tested the influence of SS-GAG on the proliferation activity of endothelial cell oxidative damage by the means of MTT assay.3. In order to observe the effect of SS-GAG on endothelial cell oxidative damage induced by oxidized low density lipoprotein (OX-LDL), 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.4. we used chemical methods to analysis the content of nitric oxide (NO) and the activity of endothelial nitric oxide synthase (eNOS) in order to study the effect of SS-GAG on endothelial cell oxidative damage induced by oxidized low density lipoprotein (OX-LDL).5. By the means of radio-immunity, we observed the action of SS-GAG on Prostacyclin-F1a (PGF1a) and Thromboxane-B2 (TXB2) contents of the endothelial cells oxidative injured by oxidized low density lipoprotein (OX-LDL).Results: 1. In model groups damaged by oxidized low density lipoprotein (OX-LDL), when compared with normal control groups, the cell number and proliferation activity of endothelial cells remarkably decreased (p<0.01); While pretreated by SS-GAG ( terminal concentration were 50ug/ml,100ug/ml, 200ug/ml) , compared with model groups the cell number and proliferation activity of endothelial cells obviously increased (p<0.01); In high dose (terminal concentration were 200ng/ml~1600ug/ml) SS-GAG protective groups, the cell number and proliferation activity were higher than those in low dose (terminal concentration was 50ug/ml) protective groups (p<0.05), and the cell number and proliferation activity in some high groups (terminal concentration were 400ug/ml~ 1600ug/ml) were even higher than normal control groups(p<0.05).2. Compared with normal control groups, in model groups damaged by hydrogen peroxide (H2O2), the proliferation activity of endothelial cells decreased remarkably (p<0.01), but it increased obviously when pretreated by SS-GAG (terminal concentration were 50ng/ml, 100ug/ml, 200ug/ml) (p<0.01). In high dose (terminal concentration was 200ug/ml) SS-GAG protective group, the proliferation activity was higher than that in low dose SS-GAG (terminal concentration was 50ug/ml) protective group (p<0.05).3. SOD testing result showed that the SOD activity of model group was much lower than that in normal control group (p<0.01); While the groups pretreated by SS-GAG (terminal concentration were 50ug/ml, 100ug/ml, 200ug/ml) , compared with model group the SOD activity obviously increased (p<0.01). MDA testing- result showed that MDA level of model group was higher than that in normal control group (p<0.01); and the groups pretreated by SS-GAG(terminal concentration were 50ug/ml, 100ug/ml, 200ug/ml) , compared with model group the MDA level obviously decreased (p<0.01).4. Compared with normal control group, the level of NO and the activity of eNOS testing results showed that NO level and eNOS activity of positive control group were lower (p<0.01); While the groups pretreated by SS-GAG(terminal concentration were 50ug/ml, 100ug/ml, 200ug/ml) , the level of NO and the activity of eNOS obviously increased (p<0.01).5. PGF1a and TXB2 testing results showed that PGF1a level of model groups much lower than normal control group (p<0.01); While the...