Adenine nucleotide and calpain inhibitor I protect against atractyloside-induced toxicity in rat renal cortical slices in vitro

Abstract

Atractyloside is a compound with a documented nephrotoxicity. It induces renal tubular necrosis at high doses and apoptosis at lower doses. This study investigates the potential protective effect of some chemical agents against atractyloside-induced nephrotoxicity in vitro using the precision-cut rat renal cortical slices obtained from kidneys of Wistar rats. For co-incubation experiments, slices were incubated for 3 h at 37 degrees C on a rocker platform with various chemical agents: ADP (5 mM), calpain inhibitor I (CPI, 1 mM), stevioside (STV, 2.5 mM) or probenecid (PRB, 2.5 mM) in the presence or absence of atractyloside (2 mM). For pre-incubation experiments, slices were incubated with the same chemical agents for 1 h before exposure to atractyloside. The nephrotoxic effects of atractyloside (2 mM) alone were manifested in several ways: by a marked increase in lactate dehydrogenase (LDH) and alkaline phosphatase (ALP) leakage, significant inhibition of p-aminohippurate (PAH) accumulation, marked depletion of intracellular ATP and reduced glutathione (GSH), and a significant reduction in pyruvate-stimulated gluconeogenesis. Co-incubation of slices with ADP or CPI and atractyloside completely blocked atractyloside-induced increase in LDH leakage, but not ALP leakage. Atractyloside-induced depletion of ATP and reduced gluconeogenesis was prevented by co-incubation with ADP or CPI. Furthermore, co-incubation of slices with STV and atractyloside, but not PRB, completely abolished atractyloside-induced depletion of ATP and decreased gluconeogenesis in the slices. Pre-incubation of slices with either ADP or CPI protected against atractyloside-induced increase in LDH leakage, reduced ATP and decreased gluconeogenesis. PAH uptake in the slices was inhibited by atractyloside and PRB in a time-dependent manner. While ADP and CPI were found to exert complete protection against atractyloside-induced toxicity irrespective of treatment schedule, STV is effective only under certain conditions, and PRB offer no protection at all. The results of this study demonstrate the usefulness of renal cortical slices as toxicology tool for evaluating and screening compounds for their potential protective effects, and are supportive of a role of adeninine nucleotide (ADP) and protease inhibitor (CPI) in protecting against atractyloside-induced cell injury.

Occurrence, Biological Activities and Synthesis of Kaurane Diterpenes and their Glycosides

“…(Asteraceae) [118], is the main compound responsible for the lethality of these plants in bovine cattle from some regions of Argentina, Brazil and Uruguay, causing hypoglycemia, respiratory depression, nephrotoxicity, hypoxemia and cell injury. In humans, the poisoning with this glycoside can cause renal and hepatic failure, also followed by necrosis [119,120]. This toxic action is due, mainly, to the inhibition of mitochondrial oxidative phosphorylation [117].…”

Atractylis gummifera L. poisoning: an ethnopharmacological review

“…Hence they cannot be used for therapeutic purposes. In another study, Obatomi et al, (2001b) compared the role of ADP, calpain inhibitor I (CPI), stevioside and probenecid in protecting against ATR-induced toxicity in rat renal cortical slices. ADP, CPI and stevioside prevented the depletion of ATP and the reduction in gluconeogenesis, whereas probenecid gave no protection at all.…”

Direct and indirect targets of carboxyatractyloside, including overlooked toxicity toward nucleoside diphosphate kinase (NDPK) and mitochondrial H + leak

“…1998 ; Obatomi et al. 1998 , 2001 ; Stewart and Steenkamp 2000 ; Daniele et al. 2005 ; Turgut et al.…”

“…Researchers have hypothesized that some regioselective organic ion transport systems (carrier protein-dependent), expressed in renal proximal tubule cells, participate in the traversal of the cell membrane by ATR/CATR (Krejci and Koechel 1992 ; Koechel and Krejci 1993 ; Obatomi and Bach 1996b ; Obatomi et al. 1998 , 2001 ; Daniele et al. 2005 ; Turgut et al.…”

“…1967 ; Stewart and Steenkamp 2000 ; Obatomi et al. 2001 ; Klingenberg 2008 ). Moreover, the structures of ATR/CATR are similar to that of oligomycin ( Figure 1 ), an antibiotic that hampers OXPHOS but directly inhibits F O F 1 -ATP synthase/ATPase ( Figure 4 ) (Al Maruf et al.…”