Studies On The Role And Its Intracellular Signal Transduction Mechanism Of Postasphyxial-serum Of Neonate In Inducing Injury Of Human Renal Tubular Cells(HK-2)

objective: To Investigate the Role of Postasphyxial-serum ofNeonate in Inducing Injury of HK-2 Cells.Methods:(1)Human renal proximal tubular cell (HK-2 cell) was used astarget cell .(2)The neonatal different concentration postasphyxial-serum of 1,3,7day after asphyxia were used as attacking means.(3)The experimental groupswere divided into 15 groups: the 2.5%,5%,10%,20% attacking concertrationgroups of 1,3,7 day after asphyxia and control group of each concertration .Theculture medium and concertration of the control group and the experimentalgroups were the same.(4)The following parameters were detected in everygroup:the changes of morphology were observed under inverted microscope,thecell viability was measured by MTT methods,and the leakage rate oflactate dehydrogenase(LDH) was determined by biochemical methods.(5)Allparameters were expressed as the mean values±standard difference( x ±S).statistical analysis between two groups were carried out by the use of student"st-test and chi-square test.Differences were considered significance when the pvalue was less than 0.05.Results: (1) Under inverted microscopy,HK-2 cells inthe experimental groups grew slowly,shank partly,the refraction rate wasstrengthened,and the quantity of cells was less than the control group.At the20% attacking concertration group of 24 hours after asphyxia,those changes inmorphology were most serious and obvious.While HK-2 cell in control groupsticked to each other tightly and grew very fast.The form and quantity of HK-2cell was normal.(2)Compared with control groups,the cell count and cellviability were lower(p<0.05).The survival rate of cell was decreased obviouslyand reached the bottom(0.172±0.039) in the 20% concentration groups of 24hours after asphyxia.There were significant differences between everyexperimental groups.(3)Compared with controls,the leakage rate of LDH in theexperimental groups were significantly increased(p<0.05) and reached thepeak(94.47±3.819) in 20% attacting concentration group of 24 hours afterasphyxia.There were significant differences between every experimental groups.Conclusion :The postasphyxial-serum of neonate is a stimulus that mayinduce injury of HK-2 cells and those injuries were most serious and obvious at24 hours after asphyxia.This deplication of the cell model offered theexperimental base to further investigation on the mechanism and prevention ofpostasphyxial renal injury in neonate.