| Objective To investigate the protective effects and mechanisms of soybean isoflavone(SI) on vascular endothelial cell(VEC) following the damage of oxidized low-density lipoprotein(ox-LDL) in vitro.Methods 1. The oxidative injury model of VEC in Vitro: Human umbilical vascular endothelial cell(HUVECs) were cultured in vitro and ox-LDL was obtained by copper-mediated oxidation. The ox-LDL with the different concentration of malondialdehycle(MDA) was added into the VECs. They were divided into 7 groups. They were control group and ox-LDL 1-6 groups (the contents of MDA were 0.2, 0.5, 0.8,1.0,1.2,1.5nmol/ml, respectively). After 24h, the contents protein and the level of MTT(OD) of cell in each group were measured. Moreover, the contents of cell MDA in control and ox-LDL 3,4,5 groups and the morphology of VECs were detected and observed by the light microscope, respectively. 2. Protective effects of SI on VEC following oxidative injury in vitro: (1) Experiment of SI incubation with normal VEC: VECs were divided into 7 groups. They were control group and the group of normal cell with different dose of SI (the contents of SI were 5, 10, 25, 50, 100, 200 #mol/L, respectively) . After 24h, the contents of cell MDA, the activity of cell superoxide-dismutase(SOD) and glutathione-peroxidase(GSH-PX) were measured. (2) Experiment of SI incubation with LDL, ox-LDL in vitro: (1)The prohibitive effect of SI on copper-mediated oxidation of LDL: The experiment included nonoxide-LDL(N-LDL)group and the group of (Cu2++LDL) with different dose of SI (the contents of SI were 0, 5, 10, 25, 50, 100, 200 # mol/L, respectively) . The time of reaction was 24h. The contents of protein and MDA were measured interval 4h. (2)The antioxidant action of SI. on ox-LDL: The experiment included N-LDL group and the group of ox-LDL with different dose of SI (the contents of SI were 0, 5, 10, 25, 50, 100, 200 # mol/L, respectively) . The contents of protein and MDA were measured after 24h. (3) The protective effects of SI on VEC following oxidative injury in vitro: VECs were divided into6 groups. They were control, ox-LDL, VE(50 # mol/L)+ox-LDL , (SI-L, M, H) (10, 50, l00# mol/L) +-ox-LDL groups, respectively. VECs were incubated with ox-LDL for 24 hours after they were incubated with VE and different dose of SI for 24 hours. The activity and antioxidant indices of cell were measured. Furthermore, the paste rate between monocyte and endothelial (MC-EC) and cell apoptosis index(AI) were detected, respectively. Besides, the morphology of VECs in each group was observed by the light and electronic microscope.Results 1. The oxidative injury model of VEC in Vitro: There was no significant difference between the control group and ox-LDL 1,2,3 groups in the contents of protein and the level of MTT(OD) of cell (P>0.05). The contents of protein and the level of MTT(OD) of cell in ox-LDL 4,5,6 groups were significantly lower than that in the control group, respectively (P<0.01). Besides, the morphology of VECs in ox-LDL 4,5,6 groups was worse than that in the other groups. But the VECs in the ox-LDL 4 group were better than that in ox-LDL 5,6 groups. Furthermore, there was no significant difference in the contents of cell MDA between the control group and the ox-LDL 3 group(P>0.05). Though, the contents of cell MDA in ox-LDL 4,5 groups were significantly higher than that in the control group(P<0.01). The contents of cell MDA in the ox-LDL 5 group were significantly higher than that in the ox-LDL 4 group(P<0.0l). 2. Protective effects of SI on VEC following oxidative injury in vitro: (1) Experiment of SI incubation with normal VEC: The contents of cell MDA in the groups of the different dose of SI were significantly lower than that in the control group and the activity of cell SOD was significantly higher(P<0.0l). On the other hand, there was no significant difference about the activity of cell GSH-PX among these groups (P>0.05 ). (2) Experiment of SI incubation with LDL, ox-LDL in vitro: (2) The prohibitive action of SI on copper-mediated oxidation... |
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