Effects Of Total Flavonoids Of Astragalus On Vascular Endothelial Cells Apoptosis Induced By Serum Of End Stage Renal Disease Patients

Objective:We establish the internal environment model of end stage renal disease in vitro. Observe the effects of total flavonoids of astragalus to oxidative stress end product4-hydroxy nonene aldehyde and ERS signal molecule CHOP expression in HUVECs. Further explore the mechanism of total flavonoids of astragalus reduce vascular endothelial cell apoptosis induced by serum of end stage renal disease patients with maintenance hemodialysis. For prevention and treatment cardiovascular complications in end stage renal disease patients with maintenance hemodialysis provide new theoretical basis and treatment of new targets.Methods:HUVECs cultivation in Dulbecco’s Modified Eagle’s Medium (DMEM), which contain10%fetal bovine serum (FBS),100U/mL Penicillin and100μg/mL streptomycin. Set the incubator’s environment maintained at37℃and5%CO2. According to the growth situation of HUVECs selectively replace fresh medium. Applicate0.25%trypsin to pass the grow adhering to the wall HUVECs. Before the experiment24h, change of the serum free medium, make the HUVECs synchronization. Randomly divided into blank control group (including10%FBS), normal control group (including10%healthy serum), end stage renal disease group (including10%end stage renal disease patients’serum), low dose group, medial dose group and high dose group with total flavonoids of astragalus (On the basis of end stage renal disease group respectively add0.5mg/mL,1.0mg/mL,2.0mg/mL total flavonoids of astragalus). After cultured for24hours, observed the morphological change of endothelial cells by microscopy, detected survival rate by methyl thiazolyl tetrazolium(MTT)assay, observed the morphological change of apoptosis by comet assay, detected the content of4-HNE by enzyme-linked immunosorbent assay (ELISA), examined the expression of CHOP by immunohistochemical staining.Results:Normal control group compared with blank control group, the difference of survival rate, apoptosis index,4-HNE and CHOP have no statistically significant (P>0.05). After cultured HUVECs induced by end stage renal disease patients with maintenance hemodialysis serum24hours, HUVECs occurred the typical change of apoptosis, such as volume reduced, structure tighted, cytoplasm condensed, organelles gathered, cell membrane budded and fallen off, cell nucleus pycnosis and so on. Comparing with normal control group, HUVECs survival rate was lower in end stage renal disease group (P<0.01), apoptosis index increased (P<0.01),4-HNE (P<0.05) and CHOP elevated (P<0.05). Adding total flavonoids of astragalus could significantly increase the survival rate of HUVECs (P<0.05), and could decrease the apoptosis index (P<0.05) and4-HNE (P<0.05), the actions appeared dose dependent (P<0.05), decrease the expression of CHOP (P<0.05), medial dose group was better than low dose group (P<0.01), but the difference between high dose group and middle does group was no statistically significant (P>0.05). Correlation analysis showed that there was positive correlation between4-HNE and CHOP (r=0.913, P<0.01).Conclusion:End stage renal disease patients’serum can induce HUVECs apoptosis. Total flavonoids of astragalus may inhibit endothelial cell apoptosis induced by end stage renal disease patients’serum through alleviating oxidative stress and endoplasmic reticulum stress.