Chlorogenic acid attenuates malondialdehyde levels and regulates major ferroptosis-related genes in testes of acrylamide-exposed rats

Subchronic GenX Exposure Induces Hepatic Alterations Accompanied by Changes in PPAR-Related Lipid Metabolism and Autophagy-Related Proteins in Adult Male C57BL/6J Mice: Partial Attenuation by Chlorogenic Acid

Background: 2,3,3,3-Tetrafluoro-2-(heptafluoropropoxy)propanoic acid (GenX) is a perfluoroether carboxylic acid that has been detected in drinking water sources. Its potential hepatotoxicity has raised concern, although the associated molecular alterations remain incompletely understood. Chlorogenic acid (CGA), a naturally occurring polyphenol, has been reported to affect oxidative stress and metabolic homeostasis. Methods: Adult male C57BL/6J mice were exposed to GenX (2 mg/kg/day) with or without CGA (30 mg/kg/day) by gavage for 12 weeks. AML12 cells were treated with GenX (10–800 μM) for 24 or 48 h to assess cell viability, and intracellular lipid accumulation was evaluated after exposure to 200 μM GenX for 24 h. Results: GenX exposure induced hepatomegaly, microvesicular steatosis, inflammatory cell infiltration, and a reduction in hepatic glycogen stores. It also decreased hepatic glutathione concentrations and increased hepatic malondialdehyde concentrations. Serum alanine aminotransferase, aspartate aminotransferase, total cholesterol, and triglyceride levels were elevated. In AML12 cells, GenX increased intracellular lipid accumulation, as assessed by Oil Red O staining. Transcriptomic analysis identified significant enrichment of the peroxisome proliferator-activated receptor (PPAR) signaling pathway. Consistently, GenX altered the expression of genes and proteins involved in lipogenesis, fatty acid uptake, lipid storage, and fatty acid oxidation, suggesting disturbed PPAR-related lipid metabolic regulation. Moreover, the decreased p-mTOR/mTOR ratio, increased LC3-II/I, and overexpression of Beclin1, p62, and inflammatory mediators in the GenX group might suggest changes in autophagy-related proteins and inflammatory response. CGA coadministration partially attenuated several GenX-induced hepatic alterations, including liver enlargement, hepatic lipid accumulation, lipid peroxidation, and changes in selected autophagy- and inflammation-related proteins. Conclusions: Subchronic GenX exposure-induced adverse hepatic effects might be associated with disrupted PPAR-related lipid metabolic regulation, oxidative stress, inflammatory responses, and changes in autophagy-related proteins. CGA might exert potential modulatory effects.

Antioxidant efficacy of N-acetylcysteine against acrylamide-induced toxicity in Leydig cells by modulating p53/Bax/Caspase-3 pathway.

Acrylamide (Acr) is a widely encountered environmental and dietary toxicant known to induce oxidative stress and disrupt male reproductive function. Leydig cells, due to their high metabolic activity and mitochondrial dependence, are particularly vulnerable to redox imbalance. N-acetylcysteine (Nac), a thiol-containing antioxidant and glutathione precursor, has been extensively studied for its cytoprotective properties. However, its modulatory effects on Acr-induced toxicity in Leydig cells and its pharmacodynamic interaction profile remain incompletely characterized. In this study, TM3 Leydig cells were exposed to Acr in the presence or absence of Nac. Cell viability was assessed by MTT assay, and chemical interaction profiles were evaluated using ZIP, Bliss, and Chou-Talalay combination index analyses. Oxidative stress parameters, including intracellular reactive oxygen species (ROS), lipid peroxidation, antioxidant enzyme activities (SOD, CAT, GPx), and glutathione levels, were measured. Apoptotic responses were analyzed through double fluorescence staining, RT-qPCR of apoptosis-related genes (Bax, Bcl2, Casp3, Trp53), and Western blot analysis of CASP3 protein expression. Acr exposure significantly reduced cell viability, increased ROS and lipid peroxidation levels, suppressed antioxidant defenses, and activated the mitochondrial apoptotic pathway. Nac treatment markedly improved cell viability, restored antioxidant capacity, reduced oxidative stress markers, and suppressed p53/Bax/Casp3-mediated apoptotic signaling. Combination analyses revealed an antagonistic interaction profile, indicating that Nac biologically limits Acr-induced cytotoxicity. Collectively, these findings demonstrate that Nac exerts protective effects in Leydig cells by modulating redox homeostasis and mitochondrial apoptosis, suggesting its potential as a protective regulator against Acr-induced reproductive toxicity.

Luteolin Mitigates Oxidative and Inflammatory Processes in Hepato-Renal Tissues of Rats Treated with Diethylnitrosamine and Carbon Tetrachloride

Luteolin (LT), a flavonoid known for its health benefits was investigated for possible ameliorative effects against hepato-renal toxicity induced by diethylnitrosamine and carbon tetrachloride in rats. Thirty-six male Wistar rats were assigned into six groups; One and two served as control and intoxicated groups, groups three-five were intoxicated and treated with LT at 10, 20 and 40mg/kg, respectively and group six received LT only (20mg/kg). The LT was given orally three times a week while Diethylnitrosamine (DEN) (50mg/kg) and CCl4 (0.5mL/kg) were administered intraperitoneally once a week for four consecutive weeks. Toxicants exposure significantly increased the activities of serum aminotransferases (ALT and AST) by 28% and 30%, and levels of serum urea and creatinine by 15% and 28%, respectively. The intoxication increased lipid peroxidation by 81% and 25%, and also decreased superoxide dismutase by 16% and 17%, and reduced glutathione by 31% and 25% in the liver and kidney of rats, respectively. In addition, inflammatory markers, myeloperoxidase and nitric oxide increased in the liver by 650% and 49%, and in the kidney by 367% and 11%, respectively in the intoxicated rats. Histology of liver revealed necrosis and multifocal hepatocellular coagulation while glomerular atrophy and tubular epithelial necrosis were seen in kidneys of intoxicated rats. Interestingly, LT pretreatment significantly attenuated markers of oxidative stress and inflammation in the liver and kidney of intoxicated rats. In conclusion, luteolin may serve as chemotherapeutic agent to mitigate the adverse effects of chemicals on the hepato-renal tissues of animals.

Olive leaf extract attenuates acrylamide-induced reproductive toxicity through reduced oxidative DNA damage and apoptosis associated with 8-OHdG and Caspase-3 in rats

Chrysin attenuates pentadecafluorooctanoic acid-induced hepatotoxicity in male Wistar rats via modulation of oxidative stress, inflammation, and metabolic dysfunction

Pentadecafluorooctanoic acid (PFOA), a persistent environmental pollutant, has been implicated in hepatotoxicity through mechanisms involving oxidative stress, inflammation, and metabolic dysfunction. This study investigated the hepatoprotective effects of chrysin against PFOA-induced liver injury in male Wistar rats. Twenty-five male Wistar rats were randomly divided into five groups (n = 5): control (20% dimethyl sulfoxide), PFOA (5 mg/kg), PFOA + chrysin (25 mg/kg), PFOA + chrysin (50 mg/kg), and chrysin only (50 mg/kg). Treatments were orally administered for 14 days. Serum and hepatic biomarkers of liver function, oxidative stress, inflammation, membrane integrity, and energy metabolism were evaluated using standard biochemical assays and enzyme-linked immunosorbent assay methods. PFOA exposure significantly (p < 0.05) increased serum liver enzymes (alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and lactate dehydrogenase), lipid peroxidation marker (malondialdehyde), and pro-inflammatory cytokines (interleukin 1 beta and interferon gamma), while significantly reducing antioxidant parameters (glutathione, catalase, superoxide dismutase, glutathione peroxidase, and glutathione-S-transferase, nitric oxide (NO), and membrane-bound enzyme activities, including adenosine triphosphatase (ATPase), Ca2+/Mg2+ ATPase, and 5’-nucleotidase. Chrysin treatment at both 25 and 50 mg/kg significantly (p < 0.05) ameliorated these alterations in a dose-dependent manner, with the 50 mg/kg dose showing greater protective efficacy. The findings indicate that PFOA-induced hepatotoxicity is associated with oxidative stress, inflammation, and metabolic dysfunction, whereas chrysin pre-treatment restores antioxidant defenses, attenuates inflammatory responses, and normalizes membrane-associated enzyme activities. Overall, chrysin demonstrates potent protective potential against PFOA-induced liver injury and may serve as a promising therapeutic candidate for environmental toxin-associated hepatotoxicity.

Black mulberry fruit extract attenuates valproic acid-induced toxicity by enhancing antioxidant defenses in FaO cells.

Valproic acid (VPA) is widely used to treat neurological and psychiatric disorders but may cause hepatotoxicity partly through oxidative stress and impaired cellular metabolism. This study investigated whether black mulberry (Morus nigra L.) fruit extract attenuates VPA-induced injury in rat FaO hepatoma cells. Cells were exposed to VPA acutely (500 μg/mL for 28 hours) or chronically (120 μg/mL for 4 weeks). Mulberry extract (0.1 mg/mL) was added 3 hours before acute VPA exposure and maintained throughout co-treatment or administered during the final 24 hours of chronic exposure. Cell viability, hydrogen peroxide production, antioxidant enzyme activities, hepatocellular injury markers, glucose concentration, and extracellular 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical-scavenging capacity were assessed. Compared with VPA alone, mulberry extract significantly increased glutathione peroxidase (GPx) activity under both acute and chronic conditions. During chronic VPA exposure, it also increased glutathione S-transferase (GST) activity and extracellular ABTS radical-scavenging capacity, improved cell viability, and reduced extracellular alanine aminotransferase (ALT) activity. During acute exposure, mulberry significantly reduced extracellular aspartate aminotransferase (AST) activity, whereas the improvement in cell viability was not statistically significant. Hydrogen peroxide production did not differ among acute treatment groups, and mulberry did not restore superoxide dismutase or catalase activity. These findings indicate that black mulberry fruit extract partially mitigates VPA-induced hepatocellular injury, primarily by enhancing glutathione-dependent antioxidant defenses. Further dose-response and in vivo studies are required to confirm its hepatoprotective potential.