We here investigated the anti-inflammatory activity of a polymethoxylated flavone-containing fraction (PMFF) from Citrus sinensis and of a prenylflavonoid-containing one (PFF) from Humulus lupulus, either alone or in combination (MIX). To this end, an in vitro model of inflammatory bowel disease (IBD), consisting of differentiated, interleukin (IL)-1β-stimulated Caco-2 cells, was employed. We demonstrated that non-cytotoxic concentrations of either PMFF or PFF or MIX reduced nitric oxide (NO) production while PFF and MIX, but not PMFF, also inhibited prostaglandin E₂ release. Coherently, MIX suppressed both inducible NO synthase and cyclooxygenase-2 over-expression besides NF-κB activation. Moreover, MIX increased nuclear factor erythroid 2–related factor 2 (Nrf2) activation, heme oxygenase-1 expression, restoring GSH and reactive oxygen and nitrogen species (RONs) levels. Remarkably, these effects with MIX were stronger than those produced by PMFF or PFF alone. Noteworthy, nobiletin (NOB) and xanthohumol (XTM), two of the most represented phytochemicals in PMFF and PFF, respectively, synergistically inhibited RONs production. Overall, our results demonstrate that MIX enhances the anti-inflammatory and anti-oxidative effects of the individual fractions in a model of IBD, via a mechanism involving modulation of NF-κB and Nrf2 signalling. Synergistic interactions between NOB and XTM emerge as a relevant aspect underlying this evidence.

A wealth of evidence underscores the bioactive properties of nutraceuticals and functional foods in addressing oxyinflammatory-based diseases with implications at both peripheral and central levels. Opuntia ficus-indica (OFI) is well-documented for its health-promoting attributes, though its fruit (OFIF) remains relatively understudied. Not only poses Metabolic Syndrome (MetS) cardiometabolic risks but also contributes significantly to cognitive impairment, especially in crucial brain areas such as hippocampus and hypothalamus. Following 8 weeks of HFD to induce MetS, rats received OFIF oral supplementation for 4 weeks to evaluate cognitive and affective modifications using behavioural paradigms, i.e. open field, burrowing, white-dark box, novelty-suppressed feeding, and object recognition tests. Our investigation extended to biochemical evaluations of lipid homeostasis, central and peripheral oxidative stress and neurotrophic pathways, correlating these measures together with circulating leptin levels. Our data revealed that OFIF modulation of leptin positively correlates with systemic and brain oxidative stress, with markers of increased anxiety-like behaviour and impaired lipid homeostasis. On the other hand, leptin levels reduced by OFIF are associated with improved antioxidant barriers, declarative memory and neurotrophic signalling. This study underscores OFIF neuroactive potential in the context of MetS-associated cognitive impairment, offering insights into its mechanisms and implications for future therapeutic strategies.

Metabolic disorders and brain insulin resistance (IR) are major risk factors for the development of neurodegenerative conditions. Kumquat fruit (KF) administration has demonstrated significant anti-dysmetabolic effects, improving peripheral IR in murine models of metabolic syndrome. Along these lines, this study evaluated the neuroprotective effects of KF supplementation in a model of dysmetabolism-induced neuronal damage and its ability to counteract the disruption of brain insulin signalling. To this end, biochemical and histological analysis assessed neuroapoptosis, disruption of brain insulin signalling and neuroinflammation in a model of high-fat diet (HFD)-induced neuronal damage. Our findings demonstrate, for the first time, that KF supplementation significantly counteracts HFD-induced neuroapoptosis downregulating pro-apoptotic genes (FAS-L, BIM and P27) and upregulating the anti-apoptotic ones (BDNF and BCL-2). Coherently, KF positively influenced the expression of selected genes related to Alzheimer’s Disease. Relevantly, these effects were associated to KF ability to restore brain insulin signalling by increasing insulin receptor expression, reducing IRS-1 serine phosphorylation, enhancing both AKT activation and GSK-3β inactivation. Accordingly, KF suppressed HFD-neuroinflammation, counteracting the overexpression of NF-κB and its downstream enzymatic products, iNOS and COX-2. Collectively, these findings demonstrate the neuroprotective benefits of KF administration, supporting its potential as a dietary intervention for dysmetabolic-related neurodegenerative disorders.

Current intra-articular therapies with hyaluronic acid (HA) provide symptomatic relief in joint diseases, but have limited efficacy in counteracting oxidative stress and inflammation, key drivers of cartilage degradation in rheumatoid arthritis (RA). To address this limitation, the potential of combining HA with the phytochemicals xanthohumol (XAN) and epigallocatechin-3-O-gallate (EGCG), known for their antioxidant and anti-inflammatory properties, was evaluated in a cellular model of RA (SW982 synoviocytes stimulated with interleukin-1β, IL-1β). The Chou–Talalay method demonstrated that their combination synergistically reduced reactive oxygen species (ROS) and nitric oxide (NO) levels. The “TRIPLE” combination (HA + XAN + EGCG) showed the lowest combination index and the highest dose reduction index. Compared to individual treatments, TRIPLE significantly decreased IL-1β-induced IL-6, IL-8, TNF-α, and MMP-3 levels, while increasing the levels of the anti-inflammatory cytokine IL-10. Western blot analysis revealed a marked reduction in iNOS, COX-2, and MMP-3 protein expression following TRIPLE treatment. Moreover, the combination inhibited IL-1β-induced phosphorylation of IκB and p65, thereby preventing NF-κB activation. These findings suggest that integrating XAN and EGCG into injectable HA formulations may represent a promising strategy to improve the management of joint inflammation in RA.

Herein, we adopted a dual approach combining molecular modeling and biological studies, in order to assess the interaction between four selected natural antioxidants (NAs; berberine, curcumin, astaxanthin, indicaxanthin) and tissue transglutaminase (TG2) levels both in the absence and in the presence of full native peptide of amyloid‐β (Aβ). Docking studies were performed to ascertain the binding affinity of NAs against the TG2 closed, Ca ²⁺ ‐bound closed, and open forms. In the biological investigation, the effect of berberine and curcumin treatment on TG2 in Olfactory Ensheathing Cells (OECs) exposed to Aβ(1–42) or to Aβ(25–35), a Aβ toxic fragment, or to reverse‐sequence fragment Aβ(35–25), an Aβ not toxic fragment, was tested. In addition, their effect on the percentage of cell viability and cytoskeleton marker (GFAP, vimentin and nestin) levels were evaluated. The role of berberine and curcumin on both endocellular levels of reactive oxygen species (ROS) and apoptotic pathway activation were also assessed. Our findings demonstrate that pretreatment of OECs with these NAs counteracted the Aβ‐induced upregulation of TG2, restoring its expression to control levels and preserving its predominant cytosolic localization. Furthermore, antioxidant pretreatment reinstated cell viability, normalized the expression of GFAP, vimentin, and nestin, reduced intracellular ROS accumulation, and prevented activation of the apoptotic cascade. Our findings demonstrate that integrating computational and biological approaches, enhances the identification of potent therapeutic agents and also highlights berberine and curcumin as promising candidates for the development of novel neuroprotective drugs against neurodegenerative disorders, including Alzheimer's disease.

Ferulago nodosa (L.) Boiss. is a species belonging to the Apiaceae family which grows spontaneously in the Balkan and southern Tyrrhenian areas of the Mediterranean Sea. Plants of the Ferulago genus are widely used in the ethnopharmacological field, mainly in the form of infusions, as remedies against panic attacks, to solve problems related to the gastrointestinal tract, and for ulcer complications. In this study, we assessed antioxidant and antiproliferative activities of the pressafonin‐A (PA), extracted from F. nodosa roots, following its spectroscopical characterization. Among the assays conducted, reducing capacity tests and free radical scavenging assays showed that PA can act as an antioxidant. PA also exhibited time‐ and concentration‐dependent antiproliferative activity against human cancer cell lines, including A‐375 (melanoma), HCT‐116 (colorectal carcinoma), and MCF‐7 (breast adenocarcinoma). Selectivity index (SI) was calculated using normal 16‐HBE cells. Among the cell lines tested, PA demonstrated an SI > 10 in A‐375 cells. Treatment with PA 6.2 μM (IC ₂₅ ) and 12.4 μM (IC ₅₀ ) for 24‐h induced apoptosis, as cytofluorimetrically evidenced by phosphatidylserine exposure, loss of mitochondrial membrane potential, and intracellular ROS production. Moreover, PA caused an arrest of cell cycle at the G1/S phase transition. Western blot analysis showed that PA caused an increase in protein levels of p53, p21 ^Waf1/Cip1 , cytochrome c, and BAX, along with a decrease in Bcl‐2 protein. In conclusion, natural PA demonstrated antioxidant and antiproliferative properties on A‐375 by inducing apoptosis via ROS‐dependent intrinsic mitochondrial pathway and cell cycle arrest. These results suggest that PA may have potential as a chemopreventive agent for skin diseases.