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Parkinson’s disease (PD) represents the second most common neurodegenerative disorder worldwide. So far, available treat- ments possess mere symptomatic action and severe long-term side effects, so the development of neuroprotective strategies is highly needed. It is now widely recognized that neuroinflammation is crucial in PD, especially in the disease progression. Therefore, pharmacological treatments able to modulate the pathological immune response may reveal effective in slowing disease progression. The microglial cells are the key component of the brain immune system and can be activated by adenosine through the interaction with A2AARs (A2A adenosine receptors). Indeed, the modulation of purinergic receptors has been associated with a slower degeneration of nigrostriatal dopaminergic cells in PD. Since previous findings demonstrating the ability of A2AAR antagonist 8-ethoxy-9-ethyladenine (ANR94) to protect nigrostriatal neurons from neuroinflammation in an animal model of PD (1), several ad hoc-designed A2AAR antagonists (ANR94 ana- logues) have been synthetized and tested in activated BV-2 microglial cells. To mimic neuroinflammation BV-2 cells were exposed to 100 ng/mL LPS or 0.5 mM MPP+ for 24 h. The potential anti-inflammatory activity of ANR94 analogues were evaluated by MTT assay and measuring the reactive oxygen spe- cies (ROS) levels by DCF-DA probe, while gene expression anal- yses of inflammatory cytokines such as TNFa, IL-1b and the pro-inflammatory enzymes iNOS and COX-2 were performed by RT-PCR. Interestingly, the newly synthetized compounds were more effective than the lead compound ANR94 in counteracting inflammatory damage suggesting their potential use as therapeu- tic agent to prevent/counteract PD. Of course, animal and clini- cal studies are needed to investigate their in vivo activity. This work was supported by MIUR-PRIN n. 20152HKF3Z and University of Camerino n. FPI000065. 1. Pinna A et al. (2005) Eur J Pharmacol 512, 157–164.

Neurodegenerative diseases have grown as a major health and societal challenge nowadays. These diseases share common pathological features like accumulation of misfolded proteins, mitochondrial disfunction, excitotoxicity, oxidative stress and inflammation. In the last years, neuroinflammation, caused by over-acting microglia, has emerged as a key element of the pathogenesis of these diseases. Therefore, the modulation of pro- inflammatory molecules from microglia could be a promising strategy to counteract neurodegeneration. Acmella oleracea (L.) is a medicinal plant whose wide use as traditional remedy is linked to its pharmacological properties, such as anti-inflamma- tory, antioxidant, analgesic and hepatoprotective1. These effects have been mostly related to its main secondary metabolite the alkylamide spilanthol. In this study, we investigated the anti-in- flammatory activities of a spilanthol-rich essential oil from A. oleracea (AO) in BV-2 microglial cells. Cells were treated with AO, pure spilanthol(S) or a nanoemulsion (NE), (composed by AO, ethyl oleate, and polysorbate 80) for 24 h, then exposed to LPS. All the treatments significantly increased cell viability (MTT assay) in respect to cells only exposed to LPS. Interestingly, AO and NE also reduced ROS levels (DCFH-DA assay), while S had no effect on this parameter. To verify if this protec- tion could be ascribed to an anti-inflammatory mechanism, the expression of IL-1b and TNF-a, COX-2 and iNOS was evaluated by RT-PCR. Interestingly, all the treatments reduced the expression of these inflammatory mediators. These results suggest AO as a potential therapeutic agent in neurodegenerative diseases thanks to its antioxidant and anti-inflammatory activities. Of note, this work also evidences a promising application of NE as a new technological formulation to further increase the potential of this essential oil.

BACKGROUND: The interest towards botanicals and plant extracts has strongly risen due to their numerous biological effects and ability to counteract chronic diseases development. Among these effects, chemoprevention which represents the possibility to counteract the cancerogenetic process is one of the most studied. The extracts of mushroom Meripilus giganteus (MG) (Phylum of Basidiomycota) showed to exert antimicrobic, antioxidant and antiproliferative effects. Therefore, since its effect in leukemic cell lines has not been previously evaluated, we studied its potential chemopreventive effect in Jurkat and HL-60 cell lines. METHODS: MG ethanolic extract was characterized for its antioxidant activity and scavenging effect against different radical species. Moreover, its phenolic profile was evaluated by HPLC-MS-MS analyses. Flow cytometry (FCM) analyses of Jurkat and HL-60 cells treated with MG extract (0-750μg/mL) for 24-72h- allowed to evaluate its cytotoxicity, pro-apoptotic and anti-proliferative effect. To better characterize MG pro-apoptotic mechanism ROS intracellular level and the gene expression level of FAS, BAX and BCL2 were also evaluated. Moreover, to assess MG extract selectivity towards cancer cells, its cytotoxicity was also evaluated in human peripheral blood lymphocytes (PBL). RESULTS: MG extract induced apoptosis in Jurkat and HL-60 cells in a dose- and time- dependent manner by increasing BAX/BCL2 ratio, reducing ROS intracellular level and inducing FAS gene expression level. In fact, reduced ROS level is known to be related to the activation of apoptosis in leukemic cells by the involvement of death receptors. MG extract also induced cell-cycle arrest in HL-60 cells. Moreover, IC50 at 24h treatment resulted 2 times higher in PBL than in leukemic cell lines. CONCLUSIONS: Our data suggest that MG extract might be considered a promising and partially selective chemopreventive agent since it is able to modulate different mechanisms in transformed cells at concentrations lower than in non-transformed ones.

L’elenco dei “botanicals” per i quali è ammesso, in accordo alle Linee guida ministeriali di riferimento, indicare l’effetto fisiologico “Regolarità della pressione arteriosa” sono numerosi e comprendono diverse parti di alcuni vegetali. Alcune di queste come le foglie di Olea Europea (OLE) e gli estratti di Crataegus Oxyacantha (Biancospino) sono state particolarmente studiate per gli effetti sulla pressione arteriosa e, più in generale per quelli protettivi nei confronti delle patologie cardiovascolari. Obiettivo della presente revisione sarà quindi quello di descrivere l’azione biologica degli estratti di OLE e Biancospino partendo dagli impieghi tradizionali, addentrandosi poi, quanto più possibile, nei meccanismi biologici responsabili degli effetti descritti ed arrivando a descrivere gli studi clinici che abbiano valutato tali effetti sull’uomo.

Numerous studies support the protective role of bioactive peptides against cardiovascular diseases. Cereals represent the primary source of carbohydrates, but they also contain substantial amounts of proteins, therefore representing a potential dietary source of bioactive peptides with nutraceutical activities. The analysis of wheat extracts purified by chromatographic techniques by means of HPLC-UV/nanoLC-nanoESI-QTOF allowed the identification of a signal of about 7 kDa which, following data base searches, was ascribed to a nonspecific lipid-transfer protein (nsLTP) type 2 from Triticum aestivum (sequence coverage of 92%). For the first time nsLTP2 biological activities have been investigated. In particular, in experiments with human umbilical vein endothelial cells (HUVEC), nsLTP2 displayed antioxidant and cytoprotective activities, being able to significantly decrease reactive oxygen species (ROS) levels and to reduce lactate dehydrogenase (LDH) release, generated following oxidative (hydrogen peroxide) and inflammatory (tumor necrosis factor α, interleukin-1β, and lipopolysaccharide) stimulation. The obtained promising results suggest potential protective role of nsLTP2 in vascular diseases prevention. PRACTICAL APPLICATION: nsLTP 2 peptide is resistant to proteases throughout the gastrointestinal tract and exerts antioxidant and cytoprotective activities. These characteristics could be exploited in vascular diseases prevention.

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Glycation, an endogenous process that leads to the production of advanced glycation end products (AGEs), plays a role in the etiopathogenesis of Alzheimer's disease (AD). Methylglyoxal (MG) is the most potent precursor of AGEs. It has been demonstrated that AGEs cause a reduction in brain derived neurotrophic factor protein (BDNF), whose dysregulation is related to AD development. Sulforaphane (SF), an isothiocyanate found in Cruciferous vegetables, is known for its chemopreventive, cardioprotective and neuroprotective actions. Aim of this study was to investigate the role of SF in counteracting MG induced damage in SH-SY5Y neuroblastoma cells. SF counteracted MG- induced neuronal death and apoptosis as measured by MTT assay, LDH release and caspase 3 activity. SF significantly reduced intracellular ROS and increased GSH levels in MG treated cells. SF inhibited the phosphorylation of the pro-apoptotic MAPK kinases induced by MG, increased the expression and the activity of the detoxifying glyoxalase enzymes GLO1 and GLO2 and increased BDNF expression of BDNF, demonstrating a pleiotropic role in counteracting AD. This work was supported by MIUR-FIRB (project RBAP11HSZS)

Methylglyoxal (MG) is a reactive dicarbonyl compound precursor of advanced glycation end products (AGEs) and is found in high levels in blood or tissues of diabetic models. MG has been correlated with diabetic complications such as macrovascular disease, retinopathy, nephropathy, aging, neuropathy and neurodegenerative conditions such as Alzheimer's disease (1). Neurodegeneration involves several mechanisms including mitochondrial dysfunction, abnormal protein aggregation and inflammation (2). MG is efficiently metabolized by the cytosolic glyoxalase system that is composed of two enzymes glyoxalase (GLO) 1 and GLO2 and a catalytic amount of GSH (3). Isothiocyanates are metabolites derived from the hydrolysis of glucosinolates, a class of compounds present in plants of the Brassica family. In particular, sulforaphane (SF), an isothiocyanate produced from glucoraphanin by the enzyme myrosinase, is known for its chemopreventive, anti-inflammatory, cardioprotective and neuroprotective actions (4,5,6). Aim of this study was to investigate the effects of SF on the interrelated mechanisms of biological glycation and MG-induced death in neuroblastoma SH-SY5Y cells. In particular, we focused on the signaling pathways of apoptosis and the modulation of the GLO system. Cell viability was evaluated by MTT and LDH assays, apoptotic cell death by measuring caspase 3 activity, ROS production by the 2',7'-dichlorofluorescein-diacetate assay, GSH levels by the monochlorobimane assay, MAPK activation and GLO1 expression by immunoblotting, GLO1 activity spectrophotometrically. MG induced neuronal cell apoptosis via activation of caspase 3 and pro-apoptotic MAPK-signalling pathways (JNK and p38) and reduced the activation of the pro-survival kinase Akt. Moreover MG significantly increased intracellular ROS production and decreased GSH levels. SF treatment significantly counteracted cell death and apoptosis induced by MG as measured by MTT, LDH and caspase 3 activity assays. SF partially inhibited MG induced activation of p38 MAPK and activated the pro-survival kinase Akt, whereas it did not show any effect on JNK activation. SF treatment reduced MG induced oxidative stress decreasing intracellular ROS production and increasing GSH levels. Moreover, for the first time, we demonstrated that SF is able to significantly increase GLO1 protein expression and activity, eliciting a fundamental role in MG detoxification. These findings suggest that SF possesses cytoprotective ability in the prevention of MG induced glycation involved in neurodegenerative complications of diabetic patients.