ABSTRACT
Objective To explore the influence of erythropoietin(EPO)on signal conduction mechanism of injury of renal tubular cells(HK-2) induced by postasphyxial-serum of neonates.Methods Heavy term asphyxia neonate were selected which amounted to 50 cases(28 male and 22 female)who had no obvious infectious diseases and immune suppression preparation,and their family consented to drawing their venous blood 5 mL after asphyxia 1 day,and put on centrifuge for 10 minutes with 2 500 r/min,then in aqueous bath for 30 minutes to inactivate complement at 56 ℃,and undertook filtration sterilization by micropore filter.The attacking concentration of serum was 200 mL/L which was prepared by adopting Dulbecco′s Modified Eagle Medium/Nutrient Mixture F12 culture fluid.HK-2 was used as the target cel1.Before attack,the experiment was designed as:blank control group,group of pretreatment with EPO(5?104 IU/L EPO medium preconditioned for 24 hours).After attack,at the 15 minutes,1 hour,2 hours,experiment was designed as:attacking control group and attacking group preconditioned with EPO at each time point.The nuclear translocation of nuclear factor ?B(NF-?B) was detected using indirect immunofluorescence.The amount of I-?B? change by NF-?B restraining was detected by Western blot,the results were scanned into computer,and image analysis software of Image-Pro Plus was used to analyze the strap.The strap integral optical density value represented the quantity of protein.Results Before attack,compared with bland control group,the nuclear translocation of NF-?B increased and amount of I-?B? decreased in group of pretreatment with EPO(Pa
ABSTRACT
Objective To investigate the role of erythropoietin(EPO)in relieving the injury of human renal tubular cells (HK-2) induced by postasphyxial-serum of neonates.Methods Human renal proximal tubular cell(HK-2) was used as the target cell.The experiment was designed as control group, asphyxia group,and group of pretreatment with EPO. The attacking concentration of serum was 200 mL/L,then the changes of morphology were observed under inverted microscope,and the cell viability was measured by 3-(4,5-dimethy lthiazcl-2-yl)-2,5-diphenyl-tetazolium bromide(MTT) methods,and the leakage rate of lactate dehydrogenase(LDH) was determined by biochemical methods.Results Compared with control group,the change in morphology of HK-2 was most serious and obvious,and the leakage rate of LDH increased significantly,and the cell viability decreased obviously in asphyxia group.But compared with asphyxia group,the change in morphology of HK-2 was obviously improved,and the leakage rate of LDH decreased and the cell viability increased in group of pretreatment with EPO in a dose-dependent manner except the group of 1 IU/mL.Conclusion EPO can play the role in relieving the injury of renal tubular cells induced by postasphyxial-serum in neonates.