5-hydroxy-1-methylhydantoin (NZ-419), an endogenous antioxidant, attenuates mycotoxin-induced oxidative stress in human kidney proximal tubular epithelial cells.

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.

Protective Effects of Limonene and a Nano-Liposomal Limonene Formulation Against Chlorfenapyr-Induced Renal Toxicity: Mechanistic Insights into NRF2/HO-1 and NF-κB/COX-2 Signaling, Mitochondrial Dysfunction, and Apoptosis in Rat Kidneys

Chlorfenapyr (CFP) is a widely used pesticide associated with nephrotoxicity through oxidative stress, inflammation, and mitochondrial dysfunction. This study investigated the protective effects of limonene (LM) and its nano-liposomal formulation (LM-LNPs) against CFP-induced renal injury in rats. Animals were divided into six groups—control, LM, LM-LNPs, CFP, CFP + LM, and CFP + LM-LNPs—and treated orally for 30 days. CFP exposure resulted in marked renal dysfunction, histopathological and ultrastructural damage, suppression of the NRF2/HO-1/NQO1 antioxidant pathway, depletion of endogenous antioxidants excessive generation of reactive oxygen and nitrogen species, lipid peroxidation products, and DNA damage. These changes were accompanied by activation of NF-κB/COX-2-mediated inflammation, mitochondrial respiratory impairment, disrupted energy metabolism, and induction of apoptosis. Co-treatment with LM significantly ameliorated these alterations, whereas LM-LNPs produced greater improvements in renal function, tissue architecture, redox homeostasis, mitochondrial function, and inflammatory and apoptotic signaling. Immunohistochemical analyses further confirmed enhanced NRF2 expression and reduced NF-κB immunoreactivity in LM-LNP-treated kidneys. Overall, nano-liposomal delivery enhanced the renoprotective efficacy of limonene, highlighting its potential as a therapeutic strategy against pesticide-induced kidney injury.

In vitro protective effects of cyanidin-3-O-glucoside against toxicity induced by continuous exposure to antimony(III) in intestinal epithelial cells.

Antimony (Sb), a heavy metal employed in PET manufacturing, is an emerging contaminant due to its ability to leach into food and beverages. Chronic exposure can impair intestinal health by disrupting epithelial barrier functionality and promoting oxidative stress, inflammation, and apoptosis. This study investigated the protective effects of cyanidin-3-O-glucoside (C3G), a dietary anthocyanin, against continuous exposure to low concentrations of Sb(III) during intestinal epithelial Caco-2 cell differentiation. C3G (20-40μM) attenuated barrier dysfunction, increasing TEER values and reducing fluorescein permeability, and reduced reactive oxygen species, mitigating inflammatory responses by inhibiting NF-κB activation and downregulating IL-6 and COX-2 expression. C3G alleviated endoplasmic reticulum (ER) stress, decreasing eIF2α phosphorylation and ATF4 and CHOP expression. Furthermore, it reduced apoptosis, as shown by lower caspase-3 activation, and displayed antioxidant activity, partly mediated by the Nrf2‑related pathway. Overall, C3G protects intestinal epithelial function from Sb-induced stress by modulating oxidative, inflammatory, ER stress, and apoptotic pathways.

Morin attenuates vancomycin-induced nephrotoxicity via Nrf-2/HO-1, SIRT1/PGC1α and AKT/FOXO1A pathways

Modulation of Dietary 5-Hydroxymethylfurfural-Induced Intestinal Epithelial Stress by Blueberry Polyphenols

Ultra-processed foods expose consumers to heat-induced contaminants like 5-hydroxymethylfurfural (5-HMF), which impairs intestinal homeostasis via oxidative stress and inflammation. This study quantified 5-HMF in bakery products and evaluated blueberry polyphenols’ capacity to attenuate 5-HMF-induced cellular stress in an in vitro Caco-2 intestinal model. 5-HMF levels in commercial bakery products were determined using a colorimetric assay and HPLC-UV. Differentiated Caco-2 cells were exposed to 5-HMF (0.4 mM) for 48 h, alone or co-treated with blueberry juice (15 µg/mL of phenol equivalents). ROS production, lipid accumulation, gene expression of antioxidant, lipid, and inflammatory markers, and NF-κB/ERK signaling pathways were analyzed. HPLC-UV accurately quantified 5-HMF in bakery products (exceeding 2 mg/100 g), revealing systematic overestimation by the colorimetric method. In Caco-2 cells, 5-HMF significantly increased ROS, lipid accumulation, inflammatory cytokines (NLRP3, IL1B, and IL18), and activated NF-κB and p-ERK. Co-treatment with blueberry juice reduced ROS and lipids, upregulated the NRF2 antioxidant pathway, and drastically suppressed NF-κB -mediated inflammation. 5-HMF induced markers of metabolic dysfunction, oxidative stress, and inflammation in the intestinal epithelial model. Polyphenol-rich blueberry juice attenuated 5-HMF-induced alterations at the tested concentration, suggesting a potential protective nutritional strategy against epithelial stress associated with food-processing contaminants.

Stanniocalcin-1 Overexpression Protects Porcine Intestinal Epithelial Cells Against TBHP-Induced Oxidative Stress by Preserving Mitochondrial Homeostasis

Stanniocalcin-1 (STC-1) is a multifunctional glycoprotein; however, its role in protecting intestinal epithelial cells against oxidative injury has not been completely elucidated. This study investigated the cytoprotective effects and underlying molecular mechanisms of STC-1 overexpression in porcine intestinal epithelial (IPEC-J2) cells subjected to tert-butyl hydroperoxide (TBHP)-induced oxidative stress. IPEC-J2 cells were transfected with pcDNA3.1/STC-1 prior to TBHP challenge. STC-1 overexpression markedly rescued cells from TBHP-induced cytotoxicity and cell death, and was associated with a reduced Bax/Bcl-2 ratio. Concurrently, elevated STC-1 expression dramatically suppressed intracellular reactive oxygen species and mitochondrial superoxide accumulation while preserving the mitochondrial membrane potential. These physiological improvements were accompanied by enhanced total antioxidant capacity and activities of key antioxidant enzymes. Mechanistically, STC-1 overexpression enhanced autophagic flux and promoted Pink1/Parkin-mediated mitophagy to eliminate dysfunctional mitochondria. Furthermore, STC-1 upregulation potentiated the AMPK–Nrf2/Sirt1 signaling axis and the subsequent transcriptional upregulation of mitochondrial quality control markers, including FoxO1, PGC-1α, and TFAM, under stress conditions. Collectively, these findings demonstrate that STC-1 safeguards porcine intestinal cells against oxidative injury by orchestrating a cooperative defense network encompassing ROS scavenging, mitochondrial homeostasis, and antioxidant defense amplification via the AMPK–Nrf2/Sirt1 pathway, highlighting a potential therapeutic target for preventing stress-associated intestinal disorders in piglets.