Prabal K. Chatterjee, Salvatore Cuzzocrea, Christoph Thiemermann

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Inhibitors of poly (ADP-ribose) synthetase protect rat proximal tubular cells against oxidant stress Prabal K. Chatterjee, Salvatore Cuzzocrea, Christoph Thiemermann Kidney International Volume 56, Issue 3, Pages 973-984 (September 1999) DOI: 10.1046/j.1523-1755.1999.00644.x Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 1 Poly (ADP-ribose) synthetase (PARS) activation by (A) 1 mM of hydrogen peroxide (H2O2) over time and (B) using increasing concentrations of H2O2 ().N = 6; *P < 0.05 vs. H2O2-control (▪). Kidney International 1999 56, 973-984DOI: (10.1046/j.1523-1755.1999.00644.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 2 Effect of PARS inhibitors on H2O2-mediated (1 mM for 1 hr) PARS activation. PARS inhibitors are: 3-AB (3 mM; N = 6), ISO (0.3 mM; N = 6) and Nic (3 mM; N = 6). Inactive structural analogs are: 3-ABA (3 mM; N = 6) and NicA (3 mM; N = 6). *P < 0.05 vs. control (□), +P < 0.05 vs. H2O2 control (▪). Kidney International 1999 56, 973-984DOI: (10.1046/j.1523-1755.1999.00644.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 3 Effect of increasing concentrations of H2O2 on (A) reduction in mitochondrial respiration and (B) increase in lactate dehydrogenase (LDH) released into the incubation medium.*P < 0.05 vs. control values; N = 6. Symbols are: () 60 min; (▪) 240 min. Kidney International 1999 56, 973-984DOI: (10.1046/j.1523-1755.1999.00644.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 4 Effect of increasing concentrations of desferrioxamine on H2O2-mediated (1 mM for 4 hr) (A) reduction in mitochondrial respiration and (B) increase in LDH released into incubation medium.*P < 0.05 vs. control values; N = 4. Kidney International 1999 56, 973-984DOI: (10.1046/j.1523-1755.1999.00644.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 5 Effect of increasing concentrations of catalase on H2O2-mediated (1 mM for 4 h) (A) reduction in mitochondrial respiration and (B) increase in LDH released into incubation medium.*P < 0.05 vs. control values; N = 4. Kidney International 1999 56, 973-984DOI: (10.1046/j.1523-1755.1999.00644.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 6 Effect of PARS inhibitors and their inactive structural analogs on H2O2-mediated (1 mM for 4 hr) impairment of mitochondrial respiration. (A) 3-AB (0.1–10 mM; N = 6), (B) ISO (0.01–1 mM; N = 6), (C) Nic (0.1–10 mM; N = 6), (D) 3-AB (3 mM; N = 6), ISO (0.3 mM; N = 6), Nic (3 mM; N = 6), 3-ABA. (3 mM; N = 6) and NicA (3 mM; N = 6), *P < 0.05 vs. H2O2-control (▪). Rat PT cell cultures were also incubated with 1 mM H2O2 for 4 hours, after which viability was assessed after 24 hr (E) after incubations in the presence of 3-AB (3 mM; N = 4), ISO (0.3 mM; N = 4), desferrioxamine (DEF, 3 mM; N = 4) or catalase (CAT, 3 U/ml; N = 4), *P < 0.05 vs. H2O2-control (▪). Kidney International 1999 56, 973-984DOI: (10.1046/j.1523-1755.1999.00644.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 7 Effect of PARS inhibitors and their inactive structural analogs on H2O2-mediated (1 mM for 4 hr) LDH release. PARS inhibitors used 3-AB (3 mM; N = 6), ISO (0.3 mM; N = 6) and Nic (3 mM; N = 6). Inactive structural analogs (negative controls) used 3-ABA (3 mM; N = 6) and NicA (3 mM; N = 6). *P < 0.05 vs. H2O2-control (▪). Kidney International 1999 56, 973-984DOI: (10.1046/j.1523-1755.1999.00644.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 8 Correlation between H2O2-mediated PARS activation (1 mM H2O2; 1 hr) and LDH released into the incubation medium (1 mM H2O2; 4 hr). Raw data are from 12 separate isolations; r = 0.93; *P < 0.05 vs. controls. Kidney International 1999 56, 973-984DOI: (10.1046/j.1523-1755.1999.00644.x) Copyright © 1999 International Society of Nephrology Terms and Conditions

Figure 9 Effect of PARS inhibitors, desferrioxamine and catalase on H2O2-mediated DNA single strand breakage after (A) 4 hours and (B) after 24 hours. Rat PT cell cultures were incubated with 1 mM H2O2 in the presence of 3-AB (3 mM; N = 4), ISO (0.3 mM; N = 4), desferrioxamine (DEF, 3 mM; N = 4) and catalase (CAT, 3 U/ml); N = (4), *P < 0.05 vs. H2O2-control (▪). Kidney International 1999 56, 973-984DOI: (10.1046/j.1523-1755.1999.00644.x) Copyright © 1999 International Society of Nephrology Terms and Conditions