Gender-Related Differences in Susceptibility to Acetaminophen-Induced Protein Arylation and Nephrotoxicity in the CD-1 Mouse

doi:10.1006/taap.1995.1031

Toxicology and Applied Pharmacology

Volume 130, Issue 2, February 1995, Pages 257-271

https://doi.org/10.1006/taap.1995.1031 Get rights and content

Abstract

Acetaminophen (APAP) is a commonly used analgesic and antipyretic agent which, in high doses, causes liver and kidney necrosis in man and animals. Damage in both target organs is greatly dependent upon biotransformation. However, in the CD-1 mouse only males exhibit cytochrome P450-dependent nephrotoxicity and selective protein covalent binding. The lack of renal toxicity in female mice may reflect the androgen dependence of renal CYP2E1. To study this, female mice were pretreated with testosterone propionate and then challenged 6 days later with APAP. Groups of control males and females were similarly challenged with APAP for comparison. All groups exhibited hepatotoxicity after APAP with similar glutathione (GSH) depletion, covalent binding, centrilobular necrosis, and elevation of plasma sorbitol dehydrogenase activity. By contrast, APAP-induced nephrotoxicity occurred only in males and in the females pretreated with testosterone. No nephrotoxicity was evident in APAP-challenged control females. The selective pattern of hepatic and renal protein arylation previously reported for male mice was similarly observed in testosterone-pretreated female mice. Western blot analysis of microsomes showed that testosterone increased renal CYP2E1 levels without altering hepatic CYP2E1. Testosterone pretreatment, in vivo, also resulted in increased activation of APAP in vitro in kidney microsomes with no effect on the in vitro activation of APAP in liver microsomes. These data suggest that APAP-mediated GSH depletion, covalent binding, and toxicity in the kidneys of testosterone-pretreated females results from increased APAP activation by the testosterone-induced renal CYP2E1. This further suggests that renal, rather than hepatic, biotransformation of APAP to a toxic electrophile is central to APAP-induced nephrotoxicity in the mouse.

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Assessment of the biochemical pathways for acetaminophen toxicity: Implications for its carcinogenic hazard potential
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In 2019 California's Office of Environmental Health Hazard Assessment (OEHHA) initiated a review of the carcinogenic hazard potential of acetaminophen. In parallel with this review, herein we evaluated the mechanistic data related to the steps and timing of cellular events following therapeutic recommended (≤4 g/day) and higher doses of acetaminophen that may cause hepatotoxicity to evaluate whether these changes indicate that acetaminophen is a carcinogenic hazard. At therapeutic recommended doses, acetaminophen forms limited amounts of N-acetyl-p-benzoquinone-imine (NAPQI) without adverse cellular effects. Following overdoses of acetaminophen, there is potential for more extensive formation of NAPQI and depletion of glutathione, which may result in mitochondrial dysfunction and DNA damage, but only at doses that result in cell death – thus making it implausible for acetaminophen to induce the kind of stable, genetic damage in the nucleus indicative of a genotoxic or carcinogenic hazard in humans. The collective data demonstrate a lack of a plausible mechanism related to carcinogenicity and are consistent with rodent cancer bioassays, epidemiological results reviewed in companion manuscripts in this issue, as well as conclusions of multiple international health authorities.
4-methylpyrazole protects against acetaminophen-induced acute kidney injury
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Citation Excerpt :
Immunohistochemistry studies have also demonstrated positive staining with both anti-APAP and anti-CYP2E1 antibodies in renal proximal tubules (Hart et al., 1995). Cytochrome P450-dependent nephrotoxicity and selective protein covalent binding have been observed in male CD-1 mice (Hoivik et al., 1995) and importantly, both NAPQI formation and activity of CYP2E1 have been shown in human kidney samples, which were significantly higher in males compared to females (Arzuk et al., 2018). Furthermore, histopathological analysis of renal biopsies from APAP overdose patients shows evidence of acute proximal and distal renal tubular necrosis as well as loss of tubular brush border (Björck et al., 1988; Kleinman et al., 1980).
Acetaminophen (APAP) hepatotoxicity is the most common cause of acute liver failure in the United States, and while a significant percentage of APAP overdose patients develop kidney injury, molecular mechanisms involved in APAP-induced nephrotoxicity are relatively unknown. We have shown that 4-methylpyrazole (4MP, Fomepizole) protects against APAP-induced liver injury by inhibiting reactive metabolite formation through Cyp2E1, and analysis of data from APAP overdose patients indicated that kidney dysfunction strongly correlated with severe liver injury. Since Cyp2E1 is also expressed in the kidney, this study explored protection by 4MP against APAP-induced nephrotoxicity. Male C57BL/6 J mice were treated with either 300 or 600 mg/kg APAP with or without 4MP for 2, 6 or 24 h, followed by measurement of APAP metabolism and tissue injury. Interestingly, levels of APAP and its non-oxidative metabolites were significantly higher in kidneys when compared to the liver. APAP-protein adducts were present in both tissues within 2 h, but were absent in kidney mitochondria, unlike in the liver. While GSH depletion was seen in both tissues, activation of c-jun N-terminal kinase and its translocation to the mitochondria, which is a critical feature of APAP-induced liver injury, was not detected in the kidney. Treatment with 4MP attenuated APAP oxidative metabolite generation, GSH depletion as well as kidney injury indicating its potential use in protection against APAP-induced nephrotoxicity. In conclusion, since reactive metabolite formation seems to be common in both liver and kidney, 4MP mediated inhibition of Cyp2E1 protects against APAP-induced nephrotoxicity. However, downstream mechanisms of APAP-induced nephrotoxicity seem distinct from the liver.
Estrogen receptor agonists protect against acetaminophen-induced hepatorenal toxicity in rats
2020, Life Sciences
Citation Excerpt :
On the other hand, several other studies have reported that female mice are more resistant to APAP overdose-induced liver damage due to improved detoxification of reactive oxygen metabolites (ROM) with an enhanced recovery of mitochondrial GSH in female mice [15–18]. Furthermore, it was suggested that the observed sex-dependent APAP toxicity was organ-specific, and female rats/mice were less susceptible to hepatotoxicity, but more sensitive to nephrotoxicity [19,20]. As being the dominant sex steroids of females, estrogens were defined to have pro-oxidant or anti-oxidant actions depending on target cells and estrogen receptor (ER) subtypes [21].
Acetaminophen-induced hepatorenal toxicity varies among sexes with controversial results among species. The aim was to compare the impact of sex and ovarian hormones on hepatorenal toxicity and to elucidate protective effects of estrogen and estrogen receptor (ER) agonists.
Under anesthesia, female rats underwent ovariectomy (OVX) or sham-OVX. Starting at postsurgical 40th day, OVX-rats received subcutaneously (each, 1 mg/kg/day) 17β-estradiol (E₂), ERβ-agonist (DPN) or ERα-agonist (PPT) for 10 days, while male and sham-OVX rats received vehicle for 10 days. Then, rats received either acetaminophen (3 g/kg) or saline by orogastric gavage and were decapitated at 24th h. Blood samples were obtained to measure serum ALT, AST, BUN, creatinine levels. Liver and kidney samples were obtained for histopathologic examination and for analyzing levels of luminol- and lucigenin-chemiluminescence, glutathione and myeloperoxidase activity.
Compared to their control groups, levels of AST, ALT, BUN, creatinine, hepatic and renal myeloperoxidase activity and chemiluminescence levels were increased, and hepatic glutathione level was decreased in acetaminophen-administered male groups, while ALT and hepatic chemiluminescence levels were not elevated in sham-OVX-rats. Both ER-agonists and E₂ reduced BUN, creatinine and reversed all oxidative parameters in renal tissues of OVX-rats. Additionally, ERα-agonist reversed all hepatic injury parameters, while ERβ-agonist elevated hepatic glutathione level.
Acetaminophen toxicity in female rats presented with a more preserved hepatic function, while renal toxicity was not influenced by sex or by the lack of ovarian hormones. Pretreatment with estrogen or ER agonists, via their antioxidant actions, provided protective effects on acetaminophen-induced hepatorenal toxicity.
Is oxidative stress involved in the sex-dependent response to ochratoxin A renal toxicity?
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Citation Excerpt :
However, the influence of sex in OTA-mediated kidney oxidative stress response has not been specifically studied. Indeed, this might be an important aspect to evaluate as sex differences regarding oxidative stress response have been observed in other scenarios such as after giving a high cholesterol diet to Wistar albino rats (Al-Rejaie et al., 2012), acetaminophen to CD-1 mouse (Hoivik et al., 1995) or cisplatin to Swiss albino mice (Naseem et al., 2015). Besides, Lash et al. (1998) found that the rates of S-(1,2,2-trichlorovinyl)glutathione (TCVG) formation in isolated kidney cells from male and female F344 rats were similar, but kidney cytosol and microsomes from males exhibited higher amounts of TCVG formation than the corresponding fractions from females, for both F344 rats and B6C3F1 mice.
Ochratoxin A (OTA) is a mycotoxin considered the most powerful renal carcinogen in rodents and classified as a possible human carcinogen. Though its mechanism of action is still unknown, indirect DNA reactivity mediated by oxidative stress has been hypothesized to play an important role. Moreover, large sex-differences have been observed in carcinogenicity studies, male rats being more sensitive than females. Male and female F344 rats were administered (p.o.) with bicarbonate or 0.5 mg OTA/kg b.w. for 7 days; or with bicarbonate, 0.21 or 0.5 mg OTA/kg b.w. for 21 days. Total glutathione (tGSH) and oxidized glutathione (GSSG) levels, glutathione S-transferase (GST) and superoxide dismutase (SOD) activities were analysed in kidneys. The standard alkaline comet assay was used in combination with Formamidopyrimidine-DNA glycosylase (Fpg) to detect oxidized DNA bases in kidney. No biologically relevant sex-differences were observed in all the oxidative-stress related parameters analysed. Indeed, no relevant oxidative-stress related response was observed between treated animals and controls. In accordance with the similar OTA levels and histopathological changes between both sexes observed previously in the same animals, and with other oxidative-stress related parameters measured by others, results support that there are no differences between sexes in the oxidative stress response to OTA.
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2018, Comprehensive Toxicology: Third Edition
This article describes mechanisms of toxicant-induced acute renal failure with an emphasis on the role of renal cell death. Mechanisms explaining why renal cells are susceptible to toxicant-induced injury are initially covered followed by a description of general responses of renal cells to toxicant exposures. The mechanisms of necrosis, apoptosis, and autophagy are discussed. The majority of the article discusses the role of cellular and biochemical mediators in the mechanisms of nephrotoxicity. Cellular targets of nephrotoxicity discussed include organelles such as the plasma membrane, mitochondria, endoplasmic reticulum, and the nucleus. Biochemical mediators of nephrotoxicity discussed include membrane transporters, cytochrome P-450 monooxygenases, reactive oxygen species, proteases, phospholipases, Ca^2 +, and epigenetic-mediated mechanisms. The final section of the article describes general experimental strategies for determining the mechanism of nephrotoxicity. Specific nephrotoxicants discussed include cisplatin, trichloroethylene, and some of its key metabolites, aminoglycosides, HgCl₂, cephaloridine, and model oxidants. Further, when appropriate, mechanisms of ischemia/reperfusion-, hypoxia-, and infection-based insult are discussed for comparison purposes.
Gender-specific expression of ATP-binding cassette (Abc) transporters and cytoprotective genes in mouse choroid plexus
2017, Toxicology
Citation Excerpt :
CYP2E1 is involved in the bioactiviation of APAP in the kidney. Previous studies have shown that higher renal CYP2E1 expression in male kidney contributes to greater susceptibility to APAP induced renal toxicity compared to female mice (Hoivik et al., 1995; Mazer and Perrone, 2008). Our data on the CP gender differences may provide a further step toward better understanding the impact of drugs and other organic metabolites on the brain.
The choroid plexus (CP) and blood-brain barrier (BBB) control the movement of several drugs and endogenous compounds between the brain and systemic circulation. The multidrug resistance associated protein (Mrp) efflux transporters form part of these barriers. Several Mrp transporters are positively regulated by the transcription factor nuclear factor erythroid-2-related factor (Nrf2) in liver. The Mrps, Nrf2 and Nrf2-dependent genes are cytoprotective and our aim was to examine basal gender differences in expression of Mrp transporters, Nrf2 and Nrf2-dependent genes (Nqo1 and Ho-1) in the brain-barriers. Previous studies have shown higher expression of Mrp1, Mrp2 and Mrp4 in female mouse liver and kidney. We hypothesized that similar renal/hepatic gender-specific patterns are present in the brain-barrier epithelia interfaces. qPCR and immunoblot analyses showed that Mrp4, Ho-1 and Nqo1 expression was higher in female CP. Mrp1, Mrp2 and Nrf2 expression in the CP had no gender pattern. Female Mrp1, Mrp2 and Mrp4 mouse brain expressions in remaining brain areas, excluding CP, were higher than male. Functional analysis of Mrp4 in CP revealed active accumulation of the Mrp4 model substrate fluo-cAMP. WT female CP had 10-fold higher accumulation in the vascular spaces than males and 60% higher than Mrp4^−/− females. Probenecid blocked all transport. Methotrexate did as well except in Mrp4^−/− females where it had no effect, suggesting compensatory induction of transport occurred in Mrp4^−/−. Collectively, our findings indicate significant gender differences in expression of Mrp transporters and cytoprotective genes in the CP and BBB.

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Regular ArticleGender-Related Differences in Susceptibility to Acetaminophen-Induced Protein Arylation and Nephrotoxicity in the CD-1 Mouse

Abstract

Regular Article
Gender-Related Differences in Susceptibility to Acetaminophen-Induced Protein Arylation and Nephrotoxicity in the CD-1 Mouse