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Aging is a multifactorial phenomenon characterized by degenerative processes closely connected to oxidative damage and chronic inflammation. Recently, many studies have shown that natural bioactive compounds are useful in delaying the aging process. In this work, we studied the effects of an in vivo supplementation of the stilbenoid pterostilbene on lifespan extension in Drosophila melanogaster. We found that the average lifespan of flies of both sexes was increased by pterostilbene supplementation with a higher effect in females. The expression of longevity related genes (Sir2, Foxo, and Notch) was increased in both sexes but with different patterns. Pterostilbene counteracted oxidative stress induced by ethanol and paraquat and up-regulated the antioxidant enzymes Ho e Trxr-1 in male but not in female flies. On the other hand, pterostilbene decreased the inflammatory mediators dome and egr only in female flies. Proteomic analysis revealed that pterostilbene modulates 113 proteins in male flies and only 9 in females. Only one of these proteins was modulated by pterostilbene in both sexes: vacuolar H[+] ATPase 68 kDa subunit 2 (Vha68-2) that was strongly down-regulated. These findings suggest a potential role of pterostilbene in increasing lifespan both in male and female flies by mechanisms that seem to be different in the two sexes, highlighting the need to conduct nutraceutical supplementation studies on males and females separately in order to give more reliable results.

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The A\(_{2A}\) adenosine receptor (A\(_{2A}\)AR) is one of the four subtypes activated by nucleoside adenosine, and the molecules able to selectively counteract its action are attractive tools for neurodegenerative disorders. In order to find novel A\(_{2A}\)AR ligands, two series of compounds based on purine and triazolotriazine scaffolds were synthesized and tested at ARs. Compound 13 was also tested in an in vitro model of neuroinflammation. Some compounds were found to possess high affinity for A\(_{2A}\)AR, and it was observed that compound 13 exerted anti-inflammatory properties in microglial cells. Molecular modeling studies results were in good agreement with the binding affinity data and underlined that triazolotriazine and purine scaffolds are interchangeable only when 5- and 2-positions of the triazolotriazine moiety (corresponding to the purine 2- and 8-positions) are substituted.

Skeletal muscle regeneration is a complex process involving crosstalk between immune cells and myogenic precursor cells, i.e., satellite cells. In this scenario, macrophage recruitment in damaged muscles is a mandatory step for tissue repair since pro-inflammatory M1 macrophages promote the activation of satellite cells, stimulating their proliferation and then, after switching into anti-inflammatory M2 macrophages, they prompt satellite cells’ differentiation into myotubes and resolve inflammation. Here, we show that acid sphingomyelinase (ASMase), a key enzyme in sphingolipid metabolism, is activated after skeletal muscle injury induced in vivo by the injection of cardiotoxin. ASMase ablation shortens the early phases of skeletal muscle regeneration without affecting satellite cell behavior. Of interest, ASMase regulates the balance between M1 and M2 macrophages in the injured muscles so that the absence of the enzyme reduces inflammation. The analysis of macrophage populations indicates that these events depend on the altered polarization of M1 macrophages towards an M2 phenotype. Our results unravel a novel role of ASMase in regulating immune response during muscle regeneration/repair and suggest ASMase as a supplemental therapeutic target in conditions of redundant inflammation that impairs muscle recovery.

Aging is a multifactorial phenomenon characterized by degenerative processes closely connected to oxidative damage and chronic inflammation. Recently, many studies have shown that natural bioactive compounds are useful in delaying the aging process. In this work, we studied the effects of an

Numerous laboratory and epidemiological studies show that the risk of developing several types of cancer can be reduced with the employment of natural substances that act with multiple mechanisms. In this context, an important role is played by the isothiocyanates. Recently, 6-(methylsulfonyl)hexyl isothiocyanate (6-MITC), present in the root of Wasabia Japonica, has stimulated the interest of researchers as a chemopreventive agent. In this particular study we have focused on evaluating 6-MITC’s in vitro cytotoxic, cytostatic and cytodifferentiating activities, as well as its pro-apoptotic potential. These effects were investigated by way of flow cytometric analysis of Jurkat and HL-60 cells as well as of healthy lymphocytes extracted from the blood of AVIS donors, in order to verify a potential selectivity of action. The results demonstrate that 6-MITC exerts a stronger cytotoxic effect on tumour cells than on healthy cells. The apoptosis induction exerted by 6-MITC on transformed cells is triggered by an extrinsic pathway, as demonstrated by the statistically significant increase in the percentage of cells with activated caspase-8. It was also observed that 6-MITC is able to limit tumour growth by slowing down and blocking the cell cycle of Jurkat and HL-60 cells respectively, in a dose- and time-related manner, while exerting no activity of any kind on the replication of healthy cells. Finally, by measuring the expression levels of CD-14 and CD-15, 6-MITC showed the ability to induce cytodifferentiation of HL-60 cells into macrophage and granulocytic phenotypes.

Neurodegenerative diseases are driven by several mechanisms such as inflammation, abnormal protein aggregation, excitotoxicity, mitochondrial dysfunction and oxidative stress. So far, no therapeutic strategies are available for neurodegenerative diseases and in recent years the research is focusing on bioactive molecules present in food. In particular, extra-virgin olive oil (EVOO) phenols have been associated to neuroprotection. In this study, we investigated the potential antioxidant and neuroprotective activity of two different EVOO extracts obtained from Quercetano cultivar trees grown in two different areas (plain and hill) of the Tuscany region (Italy). The different geographical origin of the orchards influenced phenol composition. Plain extract presented a higher content of phenyl ethyl alcohols, cinnammic acids, oleacein, oleocanthal and flavones, meanwhile, hill extract was richer in lignans. Hill extract was more effective in protecting differentiated SH-SY5Y cells from peroxide stress thanks to a marked upregulation of the antioxidant enzymes heme oxygenase 1, NADPH quinone oxidoreductase 1, thioredoxin Reductase 1 and glutathione reductase. Proteomic analysis revealed that hill extract plays a role in the regulation of proteins involved in neuronal plasticity and activation of neurotrophic factors such as BDNF. In conclusion, these data demonstrate that EVOOs can have important neuroprotective activities, but these effects are strictly related to their specific phenol composition.

Introduction: Oxidative stress has been proposed as one of the factors concurring in the pathophysiology of autoimmune thyroid diseases. Reactive oxygen species are the main expression of oxidative stress in biological systems, and their production can overcome antioxidant defenses ultimately leading to cell damage, apoptosis, and death. The present review was aimed at describing the state of the art of the relationships between oxidative stress and autoimmune thyroiditis. The most used biomarkers of oxidative stress and their correlation with thyroid function are reported. Evidence acquisition: We conducted a search of the literature in the English language starting from 2000, using the following search terms: "Hashimoto thyroiditis," "autoimmune thyroiditis," "hypothyroidism," "hyperthyroidism," "oxidative stress," "oxidants," "antioxidant," "advanced glycation end products." Both clinical studies and animal models were evaluated. Evidence synthesis: Data form clinical studies clearly indicate that the balance between oxidants and antioxidants is shifted towards the oxidative side in patients with autoimmune thyroiditis, suggesting that oxidative stress may be a key event in the pathophysiology of the disease, irrespective of thyroid function. Studies in animal models, such as the NOD.H2h4 mouse, confirm that thyroidal accumulation of ROS plays a role in the initiation and progression of autoimmune thyroiditis. Conclusions: Oxidant/antioxidant imbalance represent a key feature of thyroid autoimmunity. Oxidative stress parameters could be used as biochemical markers of chronic inflammation, to better predict the disease evolution along its natural history. Dietary habits and antioxidant supplements may provide protection from autoimmunity, opening new perspectives in the development of more tailored therapies.

Neurodegenerative diseases represent an increasingly public health problem, especially in the aging population, still in search of an effective therapy. The consumption of extra-virgin olive oil represents the main source of phenols from Mediterranean diet. In vitro and in vivo studies demonstrated preventive and protective properties of extra-virgin olive oil against neurodegeneration. In particular, the specific phenolic pattern of different extra virgin olive oils could influence their biological activity such as neuroprotection. To deepen this hypothesis, we investigated the potential differences in neuroprotective effects of four extra virgin olive oil extracts against H2O2-induced oxidative damage in differentiated SH-SY5Y cells by determining their effect in modulating antioxidant enzymes and pro-survival pathways. The different olive oil extracts were characterized for their content in tyrosol, hydroxytyrosol, oleacein and oleocanthal by HPLC. SH-SY5Y were treated with different concentrations (1-500 µg/mL) of the four extracts (corniolo, collina, pianura and spes) in the presence and absence of H2O2. Interestingly, collina is the most effective in counteracting oxidative damage as evaluated by MTT and LDH assays. Collina, pianura and spes reduced the H2O2-induced release of reactive oxygen species level as measured by dichloro-dihydro-fluorescein diacetate assay. Moreover, collina, pianura and spes increased GSH level measured by monochlorobimane assay. The extracts were also able to modulate phase II antioxidant enzymes and BDNF mRNA levels as measured by RT-PCR. Our findings confirm that extra-virgin olive oil possesses beneficial effects in counteracting neurodegeneration and these effects are strongly related to the specific pattern of olive oil phenols presents in the different olive oils. This work was supported by MIUR-PRIN 2015 (No. 20152HKF3Z)

Leaves and spiny cupules, collected from different chestnut trees, represents the subject of a multidisciplinary study aimed at characterizing the most relevant metabolites and at evaluating the potential cytoprotective effect exerted by the by-products of the chestnut grove. The potential cytoprotective effect of different extracts from chestnut leaves and spiny cupules has been evaluated by the MTT reduction assay in a microglia cellular model (BV-2), following inflammatory stimulation with LPS. Moreover, IL-1β expression induced by LPS in the presence or absence of extracts form leaves and spiny cupules has been evaluated by PCR. Preliminary results suggest a low toxicity and a promising anti-inflammatory activity of the extracts. The evaluation of the phytochemical profiles and of the nutraceutical potential of these samples represent an important step to encourage their recycling and valorization as sources of active principles, favoring the circular economy and reducing the environmental impact linked to the management of chestnut waste. This work was supported by MIUR-PRIN 2015 (No. 20152HKF3Z) and by Accademia Nazionale di Agricoltura.

Increasing experimental data suggests that the regular intake of polyphenols represents a potential way to improve the quality of life in aging. Pterostilbene is a derivative of the polyphenols resveratrol that has higher bioavailability and neuroprotective activity than resveratrol itself. The present study was carried out to investigate the anti-aging and anti-inflammatory effects of a lifelong dietary supplementation of pterostilbene in Drosophila melanogaster focusing on the expression of genes related to lifespan and senescence regulation (SIRT1; FOXO; p16; NOTCH) or to inflammatory response (DOME and Eiger1), after 2 weeks (T1) or 2 months (T2) of pterostilbene supplementation. To test the anti-neuroinflammatory effect of pterostilbene, the expression of IL1 and iNOS has been investigated by RT-PCR in BV2 microglial cells stimulated with LPS. Pterostilbene supplementation significantly increased the mean life span of both male and female fruit flies. Consistently, pterostilbene supplementation significantly increased NOTCH and SIRT1 expression in female flies at T1 and T2, whereas only at T2 in males. FOXO and p16 were up-regulated at T1 in females and at T2 in males. Eiger and DOME were reduced only in females at T1 and T2, respectively. The anti-inflammatory effect was also observed in BV-2 cells were PTS significantly reduced the up-regulation of IL1 and iNOS induced by LPS. These data confirmed the the anti-inflammatory activity of pterostilbene and suggest that it is influenced by gender.

Advanced glycation end products (AGEs) are increased in conditions of oxidative stress and promote inflammation by interacting with their receptor RAGE on cell membrane. By contrast, the soluble receptor sRAGE exerts protective effects by competing with RAGE for ligand binding. AGEs/sRAGEs interaction is involved in the pathogenesis of several diseases related to oxidative stress. In the present study, we evaluated the AGEs/sRAGEs oxidative balance in Hashimoto’ thyroiditis (HT). We measured the levels of sRAGE, by ELISA, and AGEs, by spectrophotometric method, in the serum of 50 HT patients (5 M, 45 F; mean age 38.5 ± 12 years) and 50 age-, sex- and BMI-matched healthy controls. All subjects were euthyroid at recruitment and none was on LT-4 therapy. Serum sRAGEs were significantly lower (median 424 vs 738 pg/ml; p = 0.001) and AGEs higher (205 vs 114 AU/g prot; p = 0.001) in HT patients compared to controls, and the two parameters were inversely correlated (p = 0.016). Accordingly, the AGEs/sRAGEs ratio was threefold higher in HT patients than controls (0.48 vs 0.15; p = 0.0001). In regression analysis models, serum TPO-Ab were the main predictors for AGEs and sRAGEs levels and AGEs/sRAGEs ratio (p

Background: There is a growing awareness that nutritional habits may influence risk of several inflammatory and immune-mediated disorders, including autoimmune diseases, through various mechanisms. The aim of the present study was to investigate dietary habits and their relationship with redox homeostasis in the setting of thyroid autoimmunity. Materials and Methods: Two hundred subjects (173 females and 27 males; median age, 37 years) were enrolled. None were under any pharmacological treatment. Exclusion criteria were any infectious/inflammatory/autoimmune comorbidity, kidney failure, diabetes, and cancer. In each subject, serum thyrotropin (TSH), free thyroxine, antithyroid antibodies, and circulating oxidative stress markers were measured. A questionnaire on dietary habits, evaluating the intake frequencies of food groups and adherence to the Mediterranean diet, was submitted to each participant. Results: Among the 200 recruited subjects, 81 (71 females and 10 males) were diagnosed with euthyroid Hashimoto's thyroiditis (HT); the remaining 119 (102 females and 17 males) served as controls. In questionnaires, HT subjects reported higher intake frequencies of animal foods (meat, p = 0.0001; fish, p = 0.0001; dairy products, p = 0.004) compared with controls, who reported higher intake frequencies of plant foods (legumes, p = 0.001; fruits and vegetables, p = 0.030; nuts, p = 0.0005). The number of subjects who preferentially consumed poultry instead of red/processed meat was lower in HT subjects than in controls (p = 0.0141). In logistic regression analysis, meat consumption was associated with increased odds ratio of developing thyroid autoimmunity, while the Mediterranean diet traits were protective. In HT subjects, serum advanced glycation end products (markers of oxidative stress) were significantly higher (p = 0.0001) than in controls, while the activity of glutathione peroxidase and thioredoxin reductase, as well as total plasma antioxidant activity, were lower (p = 0.020, p = 0.023, and p = 0.002, respectively), indicating a condition of oxidative stress. Stepwise regression models demonstrated a significant dependence of oxidative stress parameters on consumption of animal foods, mainly meat. Conclusions: The present study suggests a protective effect of low intake of animal foods toward thyroid autoimmunity and a positive influence of such nutritional patterns on redox balance and potentially on oxidative stress-related disorders.

Neurodegenerative diseases represent a heterogeneous group of disorders that share common features like abnormal protein aggregation, perturbed Ca2+ homeostasis, excitotoxicity, impairment of mitochondrial functions, apoptosis, inflammation, and oxidative stress. Despite recent advances in the research of biomarkers, early diagnosis, and pharmacotherapy, there are no treatments that can halt the progression of these age-associated neurodegenerative diseases. Numerous epidemiological studies indicate that long-term intake of a Mediterranean diet, characterized by a high consumption of extra virgin olive oil, correlates with better cognition in aged populations. Olive oil phenolic compounds have been demonstrated to have different biological activities like antioxidant, antithrombotic, and anti-inflammatory activities. Oleocanthal, a phenolic component of extra virgin olive oil, is getting more and more scientific attention due to its interesting biological activities. The aim of this research was to characterize the neuroprotective effects of oleocanthal against H2O2-induced oxidative stress in neuron-like SH-SY5Y cells. Moreover, protein expression profiling, combined with pathways analyses, was used to investigate the molecular events related to the protective effects. Oleocanthal was demonstrated to counteract oxidative stress, increasing cell viability, reducing reactive oxygen species (ROS) production, and increasing reduced glutathione (GSH) intracellular level. Proteomic analysis revealed that oleocanthal significantly modulates 19 proteins in the presence of H2O2. In particular, oleocanthal up-regulated proteins related to the proteasome, the chaperone heat shock protein 90, the glycolytic enzyme pyruvate kinase, and the antioxidant enzyme peroxiredoxin 1. Moreover, oleocanthal protection seems to be mediated by Akt activation. These data offer new insights into the molecular mechanisms behind oleocanthal protection against oxidative stress.

Neuroinflammation is one of the main contributors to the onset and progression of neurodegenerative diseases such as Alzheimer’s and Parkinson’s diseases. Microglial and astrocyte activation is a brain defense mechanism to counteract harmful pathogens and damaged tissues, while their prolonged activation induces neuroinflammation that can trigger or exacerbate neurodegeneration. Unfortunately, to date there are no pharmacological therapies able to slow down or stop the progression of neurodegeneration. For this reason, research is turning to the identification of natural compounds with protective action against these diseases. Considering the important role of neuroinflammation in the onset and development of neurodegenerative pathologies, natural compounds with anti-inflammatory activity could be good candidates for developing effective therapeutic strategies. Marine organisms represent a huge source of natural compounds, and among them, algae are appreciated sources of important bioactive components such as antioxidants, proteins, vitamins, minerals, soluble dietary fibers, polyunsaturated fatty acids, polysaccharides, sterols, carotenoids, tocopherols, terpenes, phycobilins, phycocolloids, and phycocyanins. Recently, numerous anti-inflammatory compounds have been isolated from marine algae with potential protective efficacy against neuroinflammation. This review highlights the key inflammatory processes involved in neurodegeneration and the potential of specific compounds from marine algae to counteract neuroinflammation in the CNS.

Different studies demonstrated that female gender is associated with improved heart failure survival, and estrogens seem to play a fundamental role in this protection. It has been suggested that 17 β-Estradiol (E2) reduces apoptosis in animal model of myocardial infarction through the modulation of MAPKs involved in oxidative stress. Moreover, recent evidences suggest that gender can influence the response to cardiovascular medications. Therefore, we hypothesized that sex hormones could also modulate the cardioprotective effects of nutraceutical compounds, such as the isothiocyanate sulforaphane (SF), present in Brassica vegetables. In a previous study we evidenced that low concentrations of SF, in the presence of E2, counteracted oxidative damage better ttna SF alone. This effect seems to be related to the up-regulation of several antioxidant enzymes (GST,HO-1,GR, Trx, NQO1) and to the activation of pro-survival pathways (Pi3K/Akt adn ERK 1/2). Aim of this study was to better clarify the mechanisms of E2-mediated potentiation of SF effects. H9c2 cardiomyoblasts were treated with SF in the absence/presence of E2 for different times. After 6h treatment, the co-treatment with SF and E2 induced a higher activation of Nrf2 transcription factor in respets to SF treatment alone. To study the involvement of E2 in SF induced Akt activation specific agonists for estrogen receptors were used. GPR30 (G protein-cooupled receptor 30) and ERβ (estrogen receptor β) emerged to be involved in Akt actovation. Moreover the co-treatment increased free [Ca++] evaluated by Fura-2 probe suggesting its involvement in E2 mediated effects. Our results increase the knowledge on E2 mediated potentiation of SF effects in cardiac cells.

Neurodegenerative diseases such as Alzheimer’s disease and Parkinson’s disease are the largest growing area of neurological research still in search of an effective therapy. Virgin olive oil is a rich source of phenolic compounds that have been demonstrated to interfere with different mechanisms involved in neurodegenerative disorders1. Olive oil polyphenols can be divided into three categories: secoiridoids such as oleocanthal, simple phenols such as hydroxytyrosol and lignanes. Here, we evaluated the neuroprotective effect of four different virgin olive oil extracts against neurodegeneration by determining their effect in modulating oxidative stress and pro-survival pathways. The different olive oil extracts were characterized for their content in tyrosol, hydroxytyrosol, oleacein and oleocanthal by HPLC. SH-SY5Y cells were treated with different concentrations (1-10 µg/mL) of the extracts. Cell viability was evaluated by MTT assay and the results showed that three of the tested extracts significantly increased cell viability. Furthermore, two of these extracts decreased basal reactive oxygen species level measured by dichloro-dihydro-fluorescein diacetate (DCFH-DA) assay. Interestingly all the extracts increased reduced GSH level measured by monochlorobimane (MCB) assay. These extracts were also able to modulate phase II antioxidant enzymes and MAPK signalling pathways as measured by RT-PCR and immunoblotting. Our findings support the idea that virgin olive oil has a beneficial health effect in counteracting neurodegeneration and these effects are strongly related to the specific pattern of olive oil phenols.

This study was carried out to characterize extracts from nine samples of the apple 'Mela Rosa dei Monti Sibillini' (MR) and to assess the antioxidant and anti-inflammatory activities. The extracts were analysed by High Performance Liquid Chromatography coupled with photodiode array detector and mass spectrometry (HPLC-DAD-MS) for 20 phytochemicals. The extracts from the lyophilized material (ELM) were richer in polyphenolic compounds than the dried ones (EDM). The MR extracts contained noteworthy amounts of the investigated analytes compared to one sample of the commercial varieties Annurca, Golden Delicious and Granny Smith used as reference. Principal component analysis (PCA) revealed that the part of the fruit seems to have a significant influence on the chemical composition of the final extract; thus, the peel extracts exhibited higher levels of phenolic compounds, especially epicatechin, procyanidin B2 and phloridzin, and triterpenes than the pulp ones. In general, the lyophilized material showed higher antioxidant activity than the dried material. The strong antioxidant capacity of the MR has also been revealed by the DPPH, ABTS, FRAP and Folin-Ciocalteau assays. The ELM of MR significantly reduced reactive oxygen species in lipopolysaccharide (LPS)-activated mouse brain microglia cells (BV-2 cells). Real-time polymerase chain reaction (RT-PCR) analysis demonstrated that the EDM and ELM of MR were effective in reducing pro-inflammatory cytokines and enzymes in BV-2 and peripheral blood mononuclear cells (PBMC). These results contribute to the exploitation of this ancient variety as a source of nutraceuticals.

Introduction Many aspects of cardiovascular diseases are similar in man and women, but some differences on risk profile, age of onset and response to medical treatments are emerging. The incidence of ischemic heart disease is significantly lower in women than in men until menopause. This observation suggests that female sex hormones, such as estrogen, may have a cardioprotective role. Our hypothesis is that sex hormones could modulate the protective effects of some nutraceutical compounds, such as the isothiocyanate sulforaphane, present in Brassica vegetables. This study was designed to explore the protective role of low concentrations of sulforaphane in the presence/absence of 17β-estradiol against oxidative stress in cardiomyocytes. Methods Primary cultures of cardiomyocytes were obtaind from 1-2 days old Sprague-Dawley rats. Cell viability was evaluated by MTT and LDH assays, ROS production by 2,7-dichlorodihydrofluorescein diacetate assay and immunofluorescence staining with anti-8-hydroxyguanosine, GSH levels by the monochlorobimane assay, antioxidant enzyme expression by RT-PCR. Transmission electron Microscopy analysis was performed to investigate the ultrastructure of cells. Results Cardiomyocytes were treated with sulforaphane and/or 17β-estradiol. Sulforaphane and 17β-estradiol co-treatment led to a higher level of protection against H2O2 induced oxidative stress than sulforaphane or 17β-estradiol alone. The co-treatment significantly reduced intracellular ROS levels in respect to 17β-estradiol or sulforaphane and increased the expression of different antioxidant enzymes in respect to sulforaphane or 17β-estradiol alone. As expected. The co-treatment increased GSH intracellular levels. Cells under oxidative stress showed a damaged cytoplasmic morphology, with several vacuoles, dilated rough endoplasmic reticulum and mitochondrial cristae. Co-treatment protected cells from oxidative stress damage, showing well preserved cytoplasmic organelles and nuclear morphology. Conclusions Our results demonstrated, for the first time, that estrogens could modify sulforaphane protective effects, suggesting that nutraceutical efficacy might be different in male and female gender.

Neurological disorders such as stroke, Alzheimer’s and Parkinson’s diseases are associated with high morbidity and mortality, and few or no effective options are available for their treatment. These disorders share common pathological characteristics like the induction of oxidative stress, abnormal protein aggregation, perturbed Ca2+ homeostasis, excitotoxicity, inflammation and apoptosis. A large body of evidence supports the beneficial effects of the Mediterranean diet in preventing neurodegeneration. As the Mediterranean diet is characterized by a high consumption of extra-virgin olive oil it has been hypothesized that olive oil, and in particular its phenols, could be responsible for the beneficial effect of the Mediterranean diet. This review provides an updated vision of the beneficial properties of olive oil and olive oil phenols in preventing/counteracting both acute and chronic neurodegenerative diseases.

The lower incidence of ischemic heart disease in female with respect to male gender suggests the possibility that female sex hormones could have specific effects in cardiovascular protection. 17β-Estradiol is the predominant premenopausal circulating form of estrogen and has a protective role on the cardiovascular system. Recent evidences suggest that gender can influence the response to cardiovascular medications; therefore, we hypothesized that sex hormones could also modulate the cardioprotective effects of nutraceutical compounds, such as the isothiocyanate sulforaphane, present in Brassica vegetables. This study was designed to explore the protective effects of sulforaphane in the presence of 17β-estradiol against H2O2-induced oxidative stress in primary cultures of rat cardiomyocytes. Interestingly, 17β-estradiol enhanced sulforaphane protective activity against H2O2-induced cell death with respect to sulforaphane or 17β-estradiol alone as measured by 3-(4,5-dimethylthiazol-2-yl)-2,5diphenyl-tetrazolium bromide and lactate dehydrogenase assays. Moreover, 17β-estradiol boosted sulforaphane ability to counteract oxidative stress, reducing intracellular reactive oxygen species and 8-hydroxy-2'-deoxyguanosine levels and increasing the expression of phase II enzymes. Using specific antagonists of estrogen receptor α and β, we observed that these effects are not mediated by estrogen receptors. Otherwise, ERK1/2 and Akt signaling pathways seem to be involved, as the presence of specific inhibitors of these kinases reduced the protective effect of sulforaphane in the presence of 17β-estradiol. Sulforaphane and 17β-estradiol co-treatment counteracted cell morphology alterations induced by H2O2 as evidenced by transmission electron microscopy. Our results demonstrated, for the first time, that estrogens could enhance sulforaphane protective effects, suggesting that nutraceutical efficacy might be modulated by sex hormones.

Glycation, an endogenous process that leads to the production of advanced glycation end products (AGEs), plays a role in the etiopathogenesis of different neurodegenerative diseases, such as Alzheimer's disease (AD). Methylglyoxal is the most potent precursor of AGEs, and high levels of methylglyoxal have been found in the cerebrospinal fluid of AD patients. Methylglyoxal may contribute to AD both inducing extensive protein cross-linking and mediating oxidative stress. The aim of this study was to investigate the role of sulforaphane, an isothiocyanate found in cruciferous vegetables, in counteracting methylglyoxal-induced damage in SH-SY5Y neuroblastoma cells. The data demonstrated that sulforaphane protects cells against glycative damage by inhibiting activation of the caspase-3 enzyme, reducing the phosphorylation of MAPK signaling pathways (ERK1/2, JNK, and p38), reducing oxidative stress, and increasing intracellular glutathione levels. For the first time, we demonstrate that sulforaphane enhances the methylglyoxal detoxifying system, increasing the expression and activity of glyoxalase 1. Sulforaphane modulated brain-derived neurotrophic factor and its pathway, whose dysregulation is related to AD development. Moreover, sulforaphane was able to revert the reduction of glucose uptake caused by methylglyoxal. In conclusion, sulforaphane demonstrates pleiotropic behavior thanks to its ability to act on different cellular targets, suggesting a potential role in preventing/counteracting multifactorial neurodegenerative diseases such as Alzheimer's.

Cardiovascular diseases (CVDs) are the major cause of death in developed countries. Oxidative stress plays a major role in the pathophysiology of cardiac disorders. Several studies have highlighted the cardinal role played by the overproduction of reactive oxygen species in the pathogenesis of ischemic myocardial damage and consequent cardiac dysfunction. Sulforaphane (SF) is a molecule within the isothiocyanate (ITC) group of organosulfur compound commonly found in cruciferous vegetables. It is reported to be capable of stimulating cellular antioxidant defenses and inducing phase 2 detoxifying enzymes, which can protect cells against oxidative damage. Although SF is known for its anticancer benefits, its role in cardioprotection is emerging. The aim of this study was to investigate the effects of SF in preventing cell damage induced by oxidative stress. Primary rat cardiomyocytes were exposed to different concentrations of SF for 24 h and subsequently treated with H2O2 to induce oxidative stress. Cell viability, and the expression of oxidative stress markers were studied. A transmission electron microscopy (TEM) analysis was carried out to evaluate the effect of SF on cell morphology. Results showed an higher cell viability and a lower level of oxidative stress in cells pre-treated with SF before peroxide exposure in respect to H2O2 treated cardiomyocytes. TEM analysis showed a well preserved morphology in cells pre-treated with SF before H2O2. These findings demonstrate that sulforaphane prevents H2O2 - induced oxidative stress and cell death in rat cardiomyocytes, suggesting a potential protective role of SF in CVDs, Italian Journal of Anatomy and Embryology, Vol. 121, No. 1 (Supplement) 2016

It has been shown that hormones can influence the response to cardiovascular medications in males and females, but no studies have been carried out on hormone-related response to nutraceuticals. This study investigated the protective effect of sulforaphane (SF) in the presence/absence of 17β-estradiol (E2) against oxidative stress in primary rat cardiomyocytes. Cells were treated with SF and/or E2 and oxidative stress induced by H2O2. SF and E2 co-treatment led to a higher level of protection against H2O2 and reduced intracellular ROS levels in respect to SF or E2 alone. Co-treatment increased GSH level and the expression of several antioxidant enzymes in respect to SF or E2. The co-treatment induced a higher activation of Akt and ERK1/2 signaling pathways in respect to SF and E2. Transmission electron microscopy analysis revealed that co-treatment was more effective in counteracting H2O2-induced alteration of cell organelles than E2 or SF. Our results demonstrate for the first time that estrogens could enhance SF protective effects, suggesting that nutraceutical efficacy might be different in males and females. This work was supported by Fondazione del Monte di Bologna e Ravenna