Protective Effects Of Astragalus Membranaceus Injection On The Cultured Human Umbilical Vein Endothelial Cells Injured By Hypoxia-Reoxygenation

Objective To investigate the protective effect of Astragalus membranaceus injection on the human umbicial vein endothelial cells (HUVECs) injured by hypoxia-reoxygenation.Methods Human umbilical veins endothelial cells(HUVECs) were cultured and go down to the future generation.After the third generation, experiments were divided into three groups: control group (n=6) ,cells were cultured under normal conditions;cells of hypoxia/reoxygenation group (n=6) were cultured in closed container of oxygen concentrations below 1% for 1 hour,and then in CO2 foster box under normal conditions for 1 hour, astragalus membranaceus injection groups ( n=6 ) ,hUVECs in astragalus groups were given different dose Astragalus membranaceus (5μg/ml,10μg/ml , 20μg/ml, 40μg/ml, 60μg/ml) injection at the time of pre-hypoxia, after 12 hours, the others and hypoxia/reoxygenation group were the same.The shape of cells were observed in phase contrast microscope.The concentrations of superoxide dismutase(SOD),Lactate dihydrogenase(LDH) activity.the content of malondialdehyde(MDA),endothelin(ET) and nitric oxide (NO) production were determined in cell medium.Results (1)The cells shrinked and became circular in hypoxia/reoxygenation group.Astragalus membranaceus inje6tion made these changes disappear or become less. The shape of cells in 40μg/ml Astragalus membranaceus injection was the same as control group(2)After treatment with hypoxia/reoxygenation, SOD activity was lower in cell medium(16.81±1.17 vs 45.44±6.26,P<0.01). Meanwhile the content of MDA(6.98 ±1.15 vs 2.38±0.49,P<0.01), LDH activity(67.58±4.72 vs 32.26±2.71,P<0.01) in cell medium and uptake rate of trypan-blue (19.42±2.62 vs 4.02±0.49,P<0.01) became higer. After treatment with Astragalus membranaceus injection, SOD activity became higer in cell medium(P<0.05 or P<0.01).Meanwhile the content of MDA ,LDH activity in cell medium were lower(P<0.05 or P<0.01).Within 5-40μg/ml concentrations of Astragalus membranaceus injection, these effects were correlated with concentrations of Astragalus membranaceus injection.But these effects did not change whenAstragalus membranaceus injection, uptake rate of trypan-blue was lower(P<0.05 or P<0.01). Moreover, the negative correlation between uptake rate of trypan-blue and concentrations of Astragalus membranaceus injection was significant(r1=-0.95,P<0.05). (3)After treatment with hypoxia/reoxygenation, the content of ET in cell medium became higer the content of ET(1463.23±46.32 vs 366.98±12.30,P<0.01). Meanwhile NO production was lower in cell medium (32.26±3.88 vs 87.41±7.63,P<0.01). After treatment with Astragalus membranaceus injection, the content of ET in cell medium .became lower(P<0.05 or P<0.01). Meanwhile NO production was higer in cell medium (P<0.01).Moreover,the positive correlation between NO production and concentrations of Astragalus membranaceus injection was significant(r2=0.93,P<0.05). the negative correlation between the content of ET and concentrations of Astragalus membranaceus injection was significant(r3=-0.97,P<0.05)Conclusion (1)The cells shrinked and became circular in hypoxia/ reoxygenation group. Astragalus membranaceus injection made these changes disappear or become less. The shape of cells in 40ug/ml Astragalus membranaceus injection was the same as control group. (2)Hypoxia/Reoxygenation could injury HUVECs correlated with lipid peroxides and lead to Hypoxia-Reoxygenation endothelial dusfunction.even led to death. Astragalus membranaceus injection can protective endothelial cellsinjured by astragalus membranaceus injection.These effects were attenuated by adminstration of different dose Astragalus membranaceus injection in range of certain dos(3) Hypoxia/Reoxygenation could injury function of HUVECs and led to endothelial dysfunction.Astragalus membranaceus injection can protective function of HUVECs...