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© Research
Publication : The Korean journal of internal medicine

Effect of t-butylhydroperoxide on p-aminohippurat uptake in rabbit renal cortical slices.

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in The Korean journal of internal medicine - 01 Jul 1994

Choi SS, Huh KD, Woo JS, Kim YK,

Link to Pubmed [PMID] – 7865484

1994 Jul; 9(2): 105-12

Oxygen free radical (superoxide radical, hydrogen peroxide, and hydroxyl radicals) have been considered to be responsible for the pathogenesis of ischemia reperfusion injury and toxic chemical injury in a variety of organs including myocardium, brain, intestine and kidneys. In in vitro models using a suspension of rat proximal tubule segments, t-butylhydroperoxide(t-BHP), a potent oxidant, induces the severity of tubular dysfunction as reflected by decreases in tubular respiration which is associated with a progressive increase in lipid peroxidation. The precise mechanism of t-BHP-induced cell injury remains to be determine. The study was carried out to determine the effect of oxygen free radicals on organic anion transport in renal proximal tubule.By renal cortical slices, we studied accumulation of organic ions, PAH efflux oxygen consumption, lactate dehydrogenase (LDH), lipid peroxidation. The data are expressed as the mean +/- SE and evaluated for significance using Student’s t-test. A probability level of 0.05 was used to establish significance.Effect of t-butylhydroperioxide(t-BHP), a potent oxidant, on organic anion p-amminohippurate(PAH) uptake was studied in rabbit renal cortical slices. t-BHP inhibited irreversibly PAH and organic cation tetraethylammonium(TEA) uptake in a dose dependent manner with IC50 of approximately 1.0 and 0.85 mM, respectively. The efflux rate constant pf PAH was not altered by the presence of 1 mM t-BHP, indicating that the inhibitory effect of t-BHP on the steady-state accumulation of PAH is due primary to the reduction in the influx of PAH across the basolateral membrane. The kinetic analysis showed that 1mM t-BHP caused a significant reduction in the maximum rate of PAH influx(Vmax) from 1.54 +/- 0.74 to 0.72 +/- 0.54 umol/g/10 min without an effect on Km, indicating that t-BHP depressed PAH influx across the basolateral membrane by reducing the number or turnover rate of active carrier for PAH transport, but not by altering substrate affinity of the carrier. Ouabain-sensitive and -insensitive oxygen consumption was not different between the control and t-BHP-treated slices. t-BHP caused an increase in LDH release and lipid peroxidation in a dose-dependent manner, which were highly correlated with changes in PAH uptake.These results suggest that t-BHP inhibition of PAH uptake is attributed to renal tubular cell damage and lipid peroxidation plays an important role in the inhibitory effect of t-BHP on PAH transport in rabbit proximl tubules.

https://pubmed.ncbi.nlm.nih.gov/7865484