E. Elmorsy

Hesperetin-Loaded PLGA Nanoparticles Ameliorate Cisplatin-Induced Oxidative Stress and Testicular Dysfunction in Rats: Association with NRF2/HO-1, NF-κB, and ACSL4/GPX4/SLC7A11 Signaling Modulation

Background: Cisplatin is a widely used chemotherapeutic agent whose gonadotoxic effects, largely driven by oxidative stress and inflammation, pose a major threat to male reproductive health. This study evaluated whether hesperetin (HES) and hesperetin-loaded poly (lactic-co-glycolic acid) nanoparticles (HES-PLGA-NPs) can protect against cisplatin-induced testicular dysfunction in adult male Sprague Dawley rats. Methods: Sixty rats were randomly assigned to six groups: control, HES, HES-PLGA-NPs, cisplatin (CIS), CIS + HES, and CIS + HES-PLGA-NPs. CIS (7.5 mg/kg/week, i.p.) was given intraperitoneally for four weeks, while HES and nano-HES (50 mg/kg/day, p.o.) were orally administered concurrently. Reproductive hormones, testicular weight, sperm parameters, oxidative stress and antioxidant markers, NRF2/HO-1 and NF-κB signaling, apoptotic indices, ferroptosis-related markers, histopathology, and GPX4/ACSL4 immunoexpression were assessed. Results: CIS caused marked reproductive dysfunction, including reduced testosterone, FSH, and LH, decreased testicular weight, and impaired sperm quality. These changes were associated with severe oxidative and nitrosative stress (↑ MDA, NO, 8-OHdG), depletion of antioxidant defenses (↓ SOD, CAT, GSH), reduced NRF2/HO-1, elevated NF-κB activity and pro-inflammatory cytokines, apoptosis-related changes (↑ Bax, caspase-3; ↓ Bcl-2), and ferroptosis-associated alterations (↑ Fe2+, TFR1, lipid ROS, ACSL4, 4-HNE; ↓ GPX4, SLC7A11). Both HES and HES-PLGA-NPs were associated with significant amelioration of these alterations, but the nanoformulation showed more pronounced protection, normalizing several oxidative and ferroptosis-associated markers toward control levels and more effectively preserving seminiferous tubule architecture and spermatogenesis. Conclusions: HES-PLGA-NPs were associated with multi-mechanistic protection against cisplatin-induced oxidative, inflammatory, apoptotic, and ferroptosis-associated damage in the testes, and may represent a promising nanoantioxidant candidate warranting further investigation for preserving male reproductive function during chemotherapy.

Citrus flavonoids mitigate the cisplatin-induced ovarian toxicity via dual modulation of Nrf2/HO-1 pathway and NF-κB axis

Cisplatin (CIS) is an effective chemotherapeutic agent whose clinical use is limited by off-target toxicity, including damage to ovarian granulosa cells, characterized by oxidative stress and inflammation. Here, the cytoprotective potential of the citrus flavonoids, naringin (NG) and hesperidin (HP), was investigated in ovarian granulosa cells exposed to CIS-induced toxicity. Molecular docking demonstrated that NG and HP exhibited strong binding affinities for key antioxidant, nuclear factor erythroid 2–related factor 2 (Nrf2) and catalase (CAT) as well as anti-apoptotic B-cell lymphoma-2 (Bcl-2) proteins, while CIS preferentially interacted with pro-apoptotic targets. In vitro, CIS dose-dependently reduced cell viability and hormone secretion (progesterone and estradiol), and these reductions were significantly restored by co-treatment with NG or HP. CIS-induced oxidative injury, marked by elevated accumulation of reactive oxygen species (ROS), lipid peroxidation, and depletion of reduced glutathione (GSH), as well as key antioxidant defenses (superoxide dismutase (SOD) and catalase (CAT), was effectively mitigated by the NG and HP. Furthermore, NG and HP suppressed the CIS-triggered inflammation by inhibiting NF-κB activation and downregulating the release of major pro-inflammatory mediators (tumor necrosis factor-alpha (TNF-α), interleukin (IL)-6, IL-8, IL-1β). They also counteracted CIS-induced apoptosis by reducing phosphorylation of protein kinase B (Akt), cytochrome c (Cyt c) release, caspase (Cas) activation, and the Bcl-2-associated X protein (Bax)/Bcl-2 ratio. These findings demonstrated that NG and HP could protect ovarian granulosa cells from CIS-induced damage through dual regulation of the Nrf2/heme oxygenase-1 (HO-1) antioxidant and nuclear factor kappa B (NF-κB) inflammatory axis, highlighting their potential as adjuvant therapies to mitigate chemotherapy-induced ovarian 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.

Enhanced hepatoprotective effects of fisetin-loaded liposomal nanoparticles against DEHP-induced liver injury in rats: modulation of oxidative stress, inflammation, apoptosis, and ferroptosis pathways.

Protective Effects of Icariin-Loaded Nanoliposomes against Methotrexate-Induced Testicular Toxicity via Modulation of NRF2/NF-κB and Ferroptosis Signaling.

Methotrexate (MTX) is a widely used chemotherapeutic and immunosuppressive agent; however, its clinical use is limited by severe testicular toxicity associated with oxidative stress, inflammation, apoptosis, and ferroptosis. Therefore, the present study investigated the protective effects of icariin-loaded nanoliposomal formulation (ICA-LNPs) against MTX-induced testicular injury in rats compared with crude icariin (ICA). Sixty adult male Sprague Dawley rats were randomly allocated into six groups: control, ICA, ICA-LNPs, MTX, MTX+ICA, and MTX+ICA-LNPs. MTX administration markedly impaired reproductive function, evidenced by significant reductions in testosterone, luteinizing hormone (LH), follicle-stimulating hormone (FSH), sperm count, motility, and viability, together with increased sperm abnormalities. MTX also disrupted the NRF2/HO-1/NQO1 signaling pathway, suppressed antioxidant defenses, and elevated oxidative stress markers. In addition, MTX significantly activated NF-κB-mediated inflammatory responses, enhanced apoptotic and ferroptotic pathways, increased iron accumulation and ACSL4 expression, and reduced GPX4 and SLC7A11 expression in testicular tissue. Histopathological examination further confirmed severe degeneration of seminiferous tubules and impaired spermatogenesis. Treatment with ICA-LNPs significantly ameliorated these alterations and demonstrated superior protective efficacy compared with crude ICA. ICA-LNPs restored reproductive hormones, improved semen quality, enhanced antioxidant status, suppressed inflammatory mediators, reduced apoptosis and ferroptosis-related markers, and preserved normal testicular histoarchitecture. Furthermore, GPX4 immunoreactivity was markedly increased, whereas ACSL4 expression was significantly reduced following ICA-LNPs treatment. In conclusion, ICA-loaded nanoliposomal formulation effectively attenuated MTX-induced testicular toxicity and was associated with reduced oxidative stress, inflammation, apoptosis, and ferroptosis, highlighting its potential as a promising therapeutic strategy for preserving male reproductive function.

Nano‐Rosmarinic Acid Protects Against Chlorfenapyr‐Induced Testicular Toxicity Through Modulation of NRF2/HO‐1 and NF‐κB/NLRP3 Signaling Pathways

Overall, RA‐loaded Chitosan nanoparticles provided superior protection against CFP‐induced testicular toxicity compared with crude RA, highlighting the potential of nanoformulation strategies to enhance RA's biological efficacy.