Research On Oxidative Damage And Death Receptor Mediated Apoptotic Mechanism Of Lead Acetate On Rat Renal Tubular Epithelial Cells

Lead is a toxic heavy metal elements, which injury multiple system and organtoxic to the human body and no physiological functions in the body, the ideal bloodlead concentration is0. Because lead accumulates in the environment for long-term,can not be degraded,enters the body through the respiratory and digestive organ,99%from the kidney metabolism, kidney is the main damaged organs of lead poisoning.Objective: To study the oxidative damage and apoptosis of rat renal tubular epithelialcells (NRK cells) induced by lead acetate and the death receptor pathway on NRKcells apoptosis induced by lead acetate. The aim of the study was to explore themechanism of animal renal toxicity induced by lead, and provide a theoretical basisfor the prevention and treatment of lead poisoning.Methods: In vitro culture NRK cells, and in the logarithmic growth phase add into0,2,10,50μM lead acetate effect after24h, determined by MTT method to detect therate of cell proliferation. Superoxide Dismutase (SOD) activity, Maleic Dialdehyde(MDA) content, Glutathione peroxidase (GSH-Px) activity and Total AntioxidantCapacity(T-Aoc) of NRK cells were measured by Xanthine Oxidase assay,Thibabituric Acid (TBA) assay, Colorimetry and ABTS method quickly,respectively;Hoechst33258and Annexin V-FITC/PI staining assay were used to detect apoptosis;The protein expression of Fas, Fasl and caspase-8were measured by western blot;Flow cytometry detected the inhibition of Z-IETD-FMK on cell apoptosis.Results: NRK cells grew well at36℃,5%CO2and the logarithmic growth phase ofNRK cells were from24h to72h;2、10、50μM lead acetate caused oxidative damagethrough significantly decreased SOD, GSH-Px activity and Total AntioxidantCapacity, increased MDA content in a dose-dependent manner; lead acetate couldinduce apoptosis in a dose-dependent manner. Study reveals lead acetate inducedapoptosis in NRK cells by death receptor pathway, mainly in:can make the Fas, Fasland caspase-8protein content increased, and the dose-effect correlation; Thecaspase-8inhibitor z-IETD-FMK significantly inhibited lead acetate inducedapoptosis in NRK cells.Conclusion: Lead acetate could inhibit the proliferation of NRK cells, Lead acetatecould inhibit the proliferation of NRK cells,cause cell oxidative damage, andapoptosis induced by via the death receptor pathway.