OBJECTIVES:Ethnobotany and the traditional use of medicinal plants is still a key starting point for the discovery of new drugs in modern medicine, despite often being underrated. Baccharis dracunculifolia DC., the major source of green propolis, has been traditionally used in South America for the treatment of malaria and parasitic infections, and for liver and gastric disorders. To investigate the molecular mechanisms involved in its immunomodulatory action, an integrated in silico/in vitro model was developed and validated. METHODS:The phytochemical profile of a hydro-ethanolic dry extract of B. dracunculifolia was investigated through UV-vis and HPLC-DAD methods and implemented with LOTUS database. Through simulated digestion and computational approaches, such as Network Pharmacology, pharmacokinetics, and biological targets were predicted. To validate bioinformatic predictions and to assess the immunomodulatory potential of B. dracunculifolia extract, in vitro tests were conducted on human monocytes (THP-1) and on peripheral blood mononuclear cells (PBMC) through ELISA dosage and flow cytometry. KEY FINDINGS:Terpenoids resulted the most enriched class in Baccharis dracunculifolia extract (BDE) (>25% w/w), followed by phenolics compounds (>15% w/w) which also included artepillin C, caffeic acid derivatives, pinocembrin, and pinobanksin. With the exception of pinocembrin, most of BDE constituents displayed a limited bioaccessibility rate. MAPK, NF-κB, several cytokines and TLR-4 were identified as key targets. These predictions were experimentally validated, as B. dracunculifolia extract, particularly 25 μg/ml, showed to upregulate MAPK ERK1/2 and p38 activation, to modulate NF-κB and to modulate cytokine release, particularly in PBMCs, where IL-1β, IL-10, IL-6, and IL-2 increased by 27%, 39%, 17%, and 21% vs control, respectively, under basal conditions, while in the inflammatory model, it caused a selective 59% reduction in Tumor necrosis factor alpha (TNF-α) vs lipopolysaccharide (LPS). This approach provided valuable guidance for experiments while reducing both cost and time. CONCLUSIONS:Our findings demonstrated that the integrated in silico/in vitro approach proved to be a valuable and cost- and time-efficient tool for studying B. dracunculifolia extract, which was shown to exert immunomodulatory activity by modulating multiple signaling pathways at both upstream and downstream levels.

Neurodegenerative diseases involve progressive neuronal loss associated with oxidative stress (OS) and inflammation. Given the limited efficacy of current therapies, natural compounds with multitarget neuroprotective potential are of growing interest. In this study, we investigated the neuroprotective effects of a standardized Magnolia officinalis (L.) bark extract (MOE) in rat brain cortical slices exposed to hydrogen peroxide-induced OS. MOE significantly recovered tissue viability and reduced ROS and malondialdehyde levels caused by OS while attenuating caspase-3, -8, and -9 activation, suggesting modulation of intrinsic and extrinsic apoptotic pathways. Shotgun proteomics using LC-HRMS/MS identified OS-induced protein expression changes reversed by MOE, with fourteen of thirty-three altered proteins rescued by MOE co-treatment. These proteins participate in several processes, including neuronal survival, OS response, and proteostasis. Bioinformatic analysis demonstrated that genes responsible for protein synthesis regulated by MOE are subjected to transcriptional regulation by factors associated with OS, including FOXO4, NRF2, and SP1. The present findings support the hypothesis that MOE exerts multitarget neuroprotective effects by modulating key proteins involved in OS responses and neuronal survival in an acute ex vivo oxidative injury model, suggesting potential relevance for mechanisms associated with NDs.

Ailanthus altissima Swingle is a tree species native to Asian countries, but its vigorous nature has led to a worldwide spread, with important ecological and social consequences. Moreover, ailanthus tree holds great potential in terms of biomass with underexplored functional value. Aiming at investigating possible applications, we profiled primary/secondary chemistry and bioactivity of leaves (AL) and flowers (AF) of A. altissima. Antioxidant constituents (phenolics, flavonoids, tannins, ascorbate), starch, pigments, free amino acids, and micro-macronutrients were quantified by spectrophotometry, HPLC–DAD, and portable XRF, respectively. Then, aqueous and hydroalcoholic extracts of AL and AF were prepared and screened for phenolics, flavonoids, triterpenes contents, antioxidant capacity (FRAP), and analysed by HPLC–DAD. Human keratinocytes (HaCaT) were employed to assess cytotoxicity and protection from H2O2-induced reactive oxygen species (ROS). The results showed that AL was higher in phenolics, flavonoids and radical-scavenging activity than AF. Amino-acid profiles diverged, with AL enriched in alanine and γ-aminobutyric acid, while AF in proline, glutamate, and aspartate, and also the mineral allocation differed (AL: Ca, S, Fe; AF: P, K, Cu, Zn). Ethanol extraction improved recovery of phenolics, flavonoids, and triterpenes and enhanced reducing power, while chromatographic separation highlighted substantial quali-quantitative discrepancies in the extracts and plant parts. When tested in HaCaT cells, all samples did not influence viability at ≤ 50 µg/mL. Moreover, hydroalcoholic extracts attenuated ROS production dose-dependently, with AF outperforming AL. Overall, A. altissima yields non-toxic antioxidant-rich extracts, which may support evidence for its management and cosmetic applications.

Several herbal and other natural products are used as ingredients in food supplements to strengthen immunity even if, very often, marketed products are proposed without a clear rationale or experimental evidence. In this study, we aimed to investigate the effect on human monocytes (THP-1) and on ex vivo human peripheral blood mononuclear cells (PBMC) of two formulations, one containing Bifidobacterium animalis subsp. lactis Bl-04® with β-glucans (for adults) and one containing Lactobacillus rhamnosus CRL1505 with elderberry extract (for children). We compared formulations with single ingredients, with bacterial lipopolysaccharide (LPS) and the drug pidotimod; cytokines expression level was evaluated testing different concentrations of samples at two exposure times. As expected, LPS caused a non-specific huge upregulation of cytokines expression both in THP-1 and in PBMC, whereas pidotimod mainly upregulated IL-2 in PBMC and IL-8 in THP-1. The two formulations showed a difference between a pro-inflammatory stimulus such as LPS, and also from an immunostimulant drug, such as pidotimod, as they mainly upregulated the expression of IL-6 and IL-10 in PBMC but not in THP-1, in a concentration-dependent mode. Probiotics were shown to play a major role, but β-glucans and elderberry extract exerted a synergistic activity. This work demonstrated that combining selected probiotics with other natural products having immunomodulatory properties is an interesting strategy to develop innovative formulations in the sector of food supplements.

Knowledge of the chemical composition of propolis is crucial for understanding the characteristics of products of different origins, but also for quality control and regulatory purposes. To date, official monographs or official analyses that allow researchers to evaluate propolis in a proper way have not yet been released. This study focuses on the characterization of twenty-seven Italian propolis samples and the identification of chemical markers that define its geographical provenance. Total polyphenol (TP) and total flavonoid (TF) content, alongside the quantification of pinocembrin, chrysin, galangin, and caffeic acid phenethyl ester (CAPE), were identified as potential markers. Additionally, DPPH assays were conducted to evaluate the antiradical activity of propolis samples. Our findings demonstrated that TPs, TFs and pinocembrin differed in propolis of different origins, especially in samples from the islands. However, the quantification of the sum of chrysin and galangin and CAPE provided a clearer distinction of the geographical origin of the propolis samples. In contrast, the DPPH assay did not prove useful for this purpose, as most results were similar and, therefore, not significant. This study lays the groundwork for future research on propolis. These findings could contribute to the development of more refined methods for distinguishing propolis origins, enhancing the understanding, valuation and quality control of this natural product in various applications.

Berberine (BER) is an alkaloid found, together with other protoberberinoids (PROTBERs), in several species used in medicines and food supplements. While some herbal preparations containing BER and PROTBERs, such as Berberis aristata DC. bark extracts, have shown promising potential for human health, their safety has not been fully assessed. Recently, the EFSA issued a call for data to deepen the pharmacokinetic and pharmacodynamic understanding of products containing BER and PROTBERs and to comprehensively assess their safety, especially when used in food supplements. In this context, new data were collected in this work by assessing: (i) the phytochemical profile of 16 different commercial B. aristata dry extracts, which are among the most widely used preparations containing BER and PROTBERs in Europe; (ii) the In Vitro and In Silico investigation of the pharmacokinetic properties of BER and PROTBERs; (iii) the In Vitro cytotoxicity of selected extracts in different human cell lines, including tests on hepatic cells in the presence of CYP450 substrates; (iv) the effects of the extracts on cancer cell migration; and (v) the In Vitro molecular effects of extracts in non-cancer human cells. Results showed that commercial B. aristata extracts contain BER as the main constituent, with jatrorrhizine as main secondary PROTBER. BER and jatrorrhizine were found to have a good bioaccessibility rate, but they interact with P-gp. B. aristata extracts showed limited cytotoxicity and minimal interaction with CYP450 substrates. Furthermore, tested extracts demonstrated inhibition of cancer cell migration and were devoid of any pro-tumoral effects in normal cells. Overall, our work provides a valuable overview to better elucidate important concerns regarding botanicals containing BER and PROTBERs.

Chronic wounds remain a significant clinical challenge, necessitating the exploration of novel therapeutic agents. Sedum telephium L. (syn. Hylotelephium telephium (L.) H. Ohba) fresh leaf juice, known for its traditional medicinal uses, was evaluated to assess its efficacy and its mechanism in promoting wound healing in vitro. Fresh leaves were collected and processed to obtain the juice, which was analyzed for polyphenols, flavonoids, polysaccharides, and proteins content. Key bioactive compounds were identified, including complex polysaccharides (2.47%), proteins (0.16%) and kaempferol and quercetin tri- di- and mono-glycosides, 0.04%, expressed as quercetin equivalent. The juice and its polysaccharidic fraction were tested on human keratinocytes (HaCaT) and fibroblasts (HFF-1) to assess cell viability, wound closure, and the production of growth factors and pro-collagen I. Our results indicated that the whole juice significantly enhanced wound closure in both cell types: +33% in keratinocytes compared to control, at 24 h, with a marked increase in fibroblast growth factor (FGF) (+21% compared to control) and LAP(TGF-β1) (+60% compared to control) and +30% in fibroblasts compared to control at 24 h, without a significant upregulation of pro-collagen I expression. The polysaccharidic fraction alone showed limited efficacy (no efficacy in keratinocytes and +20% in wound closure compared to control at 24 h in fibroblast), emphasizing the importance of the complete phytocomplex. These findings suggest that Sedum telephium L. represents a promising candidate in the setting of skin regeneration and repair.

Background: Honey is a natural product that, thanks to its composition, particularly the high sugar content, is highly appreciated as an energy nourishment. In addition to sugars, it contains many other substances (carbohydrates, free amino acids, enzymatic proteins, organic acids, polyphenols) from which the therapeutic properties of honey arise: hydrating and osmotic activity, antimicrobial action, and antioxidant and anti-inflammatory power. Objectives: The present work aims to deepen our knowledge/understanding of the activity of skin protection exerted by honey, as a synergic result of its multiple therapeutic effects. Moreover, this study wants to find possible correlations between biological properties and the chemical-physical traits of honey. Methods: To carry out this research, five varieties of citrus honey, one of acacia honey, one of chestnut honey, and one of multifloral honey were used. The honeys were first characterized by chemical-physical analysis and then were subjected to qualitative melissopalynological analysis. Tests were also carried out to evaluate both their antioxidant power and the effect on collagenase, an enzyme involved in the degradation of collagen present in the extracellular matrix and, therefore, in the processes of skin aging. Finally, honey samples were then used in in vitro experiments to assess their action in stimulating cell viability and proliferation on human keratinocytes. Results: Chemical-physical analysis demonstrated a good water content (about 17%), an important sugar content (with the monosaccharides glucose and fructose being the most represented in all the honey samples), various amino acids (with proline remarkably being the highest in all honeys), and a high concentration of polyphenols and total flavonoids (the maximum in chestnut honey, 762 mg/kg and 514 mg/kg, respectively). Conclusions The results obtained in this work confirm the ethnopharmacological use of honey in wound care, bring new scientific knowledge on the use of honey in dermatology, and highlight two fields of excellence, particularly incitrus and chestnut honey.

Objectives: This study aimed to evaluate the potential of poplar-type propolis for wound healing by analyzing its phytochemical profile and biological effects. Methods: The phytochemical characterization included specific colorimetric assays to estimate the total phenolic and flavonoid contents and the triterpenoids content. Some of the main components of poplar propolis (pinocembrin, pinobanksin, CAPE, chrysin, galangin) were analyzed using HPLC-DAD. Scavenging activity and total antioxidant capacity were assessed through DPPH and ORAC assays, respectively. Biological in vitro assays used keratinocyte, fibroblast, and monocytic cell lines. The direct wound healing properties were tested through scratching assays and ELISA kits for the assessment of the production of growth factors (FGF-7, Latency Associated Peptide - LAP), while the indirect effects were evaluated through the estimation of the levels of MMP9, IL-1β, IL-8 and TNF-α using ELISA kits together with a cell-free test on the inhibition capacity on collagenases. During the study, the Network Pharmacology analysis was employed to further explore possible mechanisms of action of propolis on the healing process. Results: The analyses confirmed the high phenolic content of poplar propolis, dominated by flavonoids, and terpenes. Propolis exhibited significant antioxidant activity and interestingly, propolis showed indirect wound-healing properties by mitigating inflammation and remodeling (reduced IL-1β and MMP9) and potentially modulating the immune response (upregulated IL-8). In vitro studies confirmed these effects, demonstrating decreased MMP9 production and collagenase inhibition when cells were co-treated with propolis and a stressor. Propolis also suppressed IL-1β release in fibroblasts, although its impact on TNF-α was inconclusive. Notably, co-treatment upregulated IL-8 in monocytes, suggesting a potential immunomodulatory role. Conclusion: poplar propolis may not directly stimulate cell proliferation during wound healing, while its anti-inflammatory and immunomodulatory properties could suggest an indirect contribution to the process.

While the number of publications on wine and health is steadily increasing, ranging from a molecular level to epidemiological studies, often with contradictory results, little attention has been given to a holistic approach to research, starting from the molecular level to arrive at pharmacological and medical conclusions. In this review, some unusual concepts are considered, such as the phytocomplex, the vehicle, and the Matrix effect. The concept of the phytocomplex is discussed, specifically the biological activities of Tyrosol, Hydroxytyrosol, and Resveratrol; indeed, the interactions among different molecules in herbal matrices provide a specific response. This is often markedly different from the response evoked by single constituents in the modulation of microbial populations in the gut, in intestinal stability and bioaccessibility, and, obviously, in inducing biological responses. Among the many alcoholic beverages which contain these molecules, wine has the most peculiar Matrix effect, which can heavily influence the bioavailability of the phytocomplex obtained by the fermentation processes that produce this beverage. Wine’s Matrix effect plays an instrumental role in improving the beneficial compounds’ bioavailability and/or in inhibiting alcohol metabolites’ carcinogenicity. Underestimation of the wine Matrix effect could lead to deceiving results, as in the case of dealcoholized wine or wine-compound-based nutritional supplements; alternatively, this can occur in the emphasis of a single component’s toxic activity, in this case, alcohol, ignoring the specific molecular-level protective action of other compounds (polyphenols) that are present in the same matrix. The dark side of the Matrix effect is also discussed. This review confirms the research recommendations made by the WHO Scientific Group, which suggests it is important “to investigate the possible protective effects of ingredients other than alcohol in alcoholic beverages”, considering that most recent studies seem not only relevant but also capable of directing future research towards innovative points of view that have so far been too neglected.

Objectives: The objective of this study is to evaluate the wound healing potential of Eurasian propolis by analyzing the phytochemical profile and the biological effects of three representative propolis samples. Methods: Specific colorimetric assays were used to estimate the total phenolic and flavonoid contents and the triterpenoids content. Some of the main components of Eurasian propolis (pinocembrin, pinobanksin, CAPE, chrysin and galangin) were analyzed using HPLC-DAD. Scavenging activity and total antioxidant capacity were assessed through DPPH and ORAC assays, respectively. Human keratinocyte, fibroblast, and monocytic cell lines were used for the biological in vitro analyses. The direct wound healing properties were tested through scratching assays and ELISA kits for the assessment of the production of growth factors (FGF-7, Latency Associated Peptide-LAP), while the indirect effects were evaluated through the estimation of the levels of MMP9, IL-1β, IL-8, and TNF-α using ELISA kits together with a cell-free test on the inhibition capacity on collagenases. Network Pharmacology analysis was employed to further explore possible mechanisms of the action of propolis on the healing process. Results: The analyses confirmed the high phenolic content of Eurasian propolis (142.50–211.30 mg GAE/g), dominated by flavonoids (95.50–196.80 mg Galangin Equivalents/g), and terpenes (431.50–650.00 mg β-sitosterol Equivalents/g), while also verifying the significant antioxidant (4.9–8.9 mM/g Trolox Equivalents) and antiradical (DPPH IC50 26.1–54.4 μg/mL) activities. The samples showed indirect wound healing properties by mitigating inflammation and remodeling (reduced IL-1β and MMP9) and potentially modulating the immune response (upregulated IL-8). In vitro studies confirmed these effects, demonstrating decreased MMP9 production and collagenase inhibition when cells were co-treated with propolis and a stressor. Propolis also suppressed IL-1β release in fibroblasts, although its impact on TNF-α was inconclusive. Notably, co-treatment upregulated IL-8 in monocytes, suggesting a potential immunomodulatory role. Conclusions: Eurasian propolis may not directly stimulate cell proliferation during wound healing. Its anti-inflammatory and immunomodulatory properties could indicate an indirect contribution in helping the process.

Berberine (BER) is a natural alkaloid found in various Berberis species, used traditionally in herbal medicine and dietary supplements. BER exhibited various biological activities, including anti-inflammatory, antioxidant, antidiabetic, and anti-tumour properties by modulating cellular pathways and influencing gene expression. While BER shows promise in various health applications, its possible toxicity raises concerns. In 2023, EFSA issued a call for data on BER- and protoberberine (PROTBER)-containing products, to comprehensively assess its safety and potential health claims. With these premises, new data was collected by assessing: i) the phytochemical profile of BER- and PROTBER-containing preparations by newly developed HPLC-DAD method; ii) in silico and in vitro investigation of pharmacokinetics properties of BER; iii) in vitro cytotoxicity in different human cell lines and, iv) biological and molecular activities of extracts containing BER and PROTBER in in vitro human non-cancer cells. By testing 16 commercial Berberis extracts, the evidence collected showed a good qualitative reliability in terms of phytochemical profile and limited relapses on cytotoxicity even at 200 µg/ml after a 24h of exposure, while evidencing a good cellular bioavailability and cell migration inhibition. The potential for plant-drug interactions affecting CYP450 proved to be minimal. Overall, our work provides a useful overview to better elucidate potential concerns on botanicals containing BER and minor PROTBER commonly used in food supplement.

The increasing use of red yeast rice (RYR) as a natural supplement to manage blood cholesterol levels is driven by its active compound, monacolin K (MK), which is chemically identical to the statin drug lovastatin (LOV). Despite its growing popularity, concerns persists regarding the safety and efficacy of RYR compared to pure statins. This study aimed to evaluate the phytochemical composition, pharmacological effects, and safety profile of various RYR samples in comparison with LOV. RYR samples with different MK content were analyzed using HPLC-DAD to quantify monacolins and other bioactive compounds. The inhibitory activity on HMG-CoA reductase was assessed through an enzymatic assay, while pharmacokinetic properties were predicted using in vitro simulated digestion and in silico models. In vitro cytotoxicity was evaluated in intestinal, hepatic, renal, and skeletal muscle cell models. Additionally, the transcriptional levels of muscle damage-related target genes were evaluated by qRT-PCR in skeletal muscle cells treated with a selection of RYR samples. Significant variability in the phytochemical composition of RYR samples was observed, particularly in the content of secondary monacolins, triterpenes, and polyphenols. The RYR phytocomplex exhibited superior inhibition of HMG-CoA reductase activity compared to isolated LOV, suggesting synergistic effects between secondary monacolins and other compounds. Molecular insights revealed that RYR samples had a lower impact on muscle cells than LOV, as reflected also by cell viability. These findings suggest that RYR could serve as a safe alternative to purified statins. However, further research is needed to fully elucidate the mechanisms behind the synergistic activity of the phytocomplex and to firmly establish the clinical efficacy of this natural product.

Background: Semen handling and cryopreservation increase reactive oxygen species (ROS) production, exposing spermatozoa to oxidative stress (OS) that can be minimised by antioxidant supplementation. We studied if Olea europaea leaf extract (OE) could have antioxidant and protective activity during sperm manipulation.Methods: The extract was characterized by high-performance liquid chromatography with diode array detection (HPLC-DAD) and antioxidant activity was determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and oxygen radical absorbance capacity (ORAC) assays. Then, spermatozoa of 35 normozoospermic donors were treated with OE. First, swim-up selected spermatozoa were incubated with OE (1:100-1:400) and sperm motility and DNA integrity (acridine orange test) were assessed. Then, swim-up selected sperm were treated with 100 mu M H2O2 to induce OS with and without OE; motility, DNA integrity and F-2-Isoprostanes (F-2-IsoPs quantified by gas chromatography/negative ion chemical ionization tandem mass spectrometry analysis), an OS marker, were assessed. Finally, sperm were frozen with and without OE. Beside the previous endpoints, acrosome shape was evaluated by Tetramethylrhodamine (TRITC)-conjugated Pisum sativum agglutinin (PSA).Results: OE was enriched in polyphenols (0.34%) and triterpenes (0.72%) and the synergistic action of phytocomplex components was responsible for antioxidant activity. Since OE was not toxic for spermatozoa, 1:100 dilution was used for the other experiments. OE protected motility, DNA integrity and reduced F-2-IsoPs (p < 0.001) in in vitro experiment with OS induction. OE treated frozen-thawed spermatozoa showed increased motility, DNA integrity, reduced F-2-IsoP concentration (p < 0.001) and normal acrosomes versus non-supplemented samples.Conclusions: OE is characterized by high concentration of polyphenols and exhibits protective properties against oxidative damage induced by H2O2 in human ejaculated sperm. After cryopreservation, the samples supplemented with the extract showed increased sperm quality. OE could represent a supplement of culture media during semen handling where OS is exacerbated.

Phaseolus vulgaris L. (common bean) contains high levels of proteins, unsaturated fatty acids, minerals, fibers, and vitamins, and for this reason, it represents an essential component of the diet. More than 40,000 varieties of beans have been recognized and are staple foods in the traditional cuisine of many countries. In addition to its high nutritional value, P. vulgaris is also characterized by its nutraceutical properties and favors environmental sustainability. In this manuscript, we studied two different varieties of P. vulgaris, Cannellino and Piattellino. We investigated the effects of traditional processing (soaking and cooking) and in vitro gastrointestinal digestion of beans on their phytochemical composition and anticancer activity. Using HT29 and HCT116 colon cancer cell lines, we showed that the extract obtained after gastrointestinal digestion of cooked beans (the bioaccessible fraction, BF) induces cell death through the induction of the autophagic process. We demonstrated that the BF of Cannellino and Piattellino beans at the concentration of 100 μg/mL reduces cell vitality, measured by MMT assay, of both HT29 (88.41% ± 5.79 and 94.38% ± 0.47) and HCT116 (86.29% ± 4.3 and 91.23% ± 0.52) cell lines. Consistently, the treatment of HT29 cells with 100 μg/mL of Cannellino and Piattellino BFs reduced clonogenicity by 95% ± 2.14 and 96% ± 0.49, respectively. Moreover, the activity of extracts appeared to be selective for colon cancer cells. The data shown in this work further confirm P. vulgaris to be among foods with beneficial effects for human health.

Except for specific vaccines and monoclonal antibodies, effective prophylactic or post-exposure therapeutic treatments are currently limited for COVID-19. Propolis, a honeybee's product, has been suggested as a potential candidate for treatment of COVID-19 for its immunomodulatory properties and for its powerful activity against various types of viruses, including common coronaviruses. However, direct evidence regarding the antiviral activities of this product still remains poorly documented. VERO E6 and CALU3 cell lines were infected with SARS-CoV-2 and cultured in the presence of 12.5 or 25 μg/ml of a standardized Hydroalcoholic Extract acronym (sHEP) of Eurasian poplar type propolis and analyzed for viral RNA transcription, for cell damage by optical and electron microscopy, and for virus infectivity by viral titration at 2, 24, 48, and 72 h post-infection. The three main components of sHEP, caffeic acid phenethyl ester, galangin, and pinocembrin, were tested for the antiviral power, either alone or in combination. On both cell lines, sHEP showed significant effects mainly on CALU3 up to 48 h, i.e., some protection from cytopathic effects and consistent reduction of infected cell number, fewer viral particles inside cellular vesicles, reduction of viral titration in supernatants, dramatic drop of N gene negative sense RNA synthesis, and lower concentration of E gene RNA in cell extracts. Interestingly, pre-treatment of cells with sHEP before virus inoculation induced these same effects described previously and was not able to block virus entry. When used in combination, the three main constituents of sHEP showed antiviral activity at the same levels of sHEP. sHEP has a remarkable ability to hinder the replication of SARS-CoV-2, to limit new cycles of infection, and to protect host cells against the cytopathic effect, albeit with rather variable results. However, sHEP do not block the virus entry into the cells. The antiviral activity observed with the three main components of sHEP used in combination highlights that the mechanism underlying the antiviral activity of sHEP is probably the result of a synergistic effect. These data add further emphasis on the possible therapeutic role of this special honeybee's product as an adjuvant to official treatments of COVID-19 patients for its direct antiviral activity.

Cannabis sativa L. is increasingly emerging for its protective role in modulating neuroinflammation, a complex process orchestrated among others by microglia, the resident immune cells of the central nervous system. Phytocannabinoids, especially cannabidiol (CBD), terpenes, and other constituents trigger several upstream and downstream microglial intracellular pathways. Here, we investigated the molecular mechanisms of a CBD- and terpenes-enriched C. sativa extract (CSE) in an in vitro model of neuroinflammation. We evaluated the effect of CSE on the inflammatory response induced by exposure to lipopolysaccharide (LPS) in BV-2 microglial cells, compared with CBD and β-caryophyllene (CAR), CB2 receptors (CB2r) inverse and full agonist, respectively. The LPS-induced upregulation of the pro-inflammatory cytokines IL-1β, IL-6, and TNF-α was significantly attenuated by CSE and only partially by CBD, whereas CAR was ineffective. In BV-2 cells, these anti-inflammatory effects exerted by CSE phytocomplex were only partially dependent on CB2r modulation and they were mediated by the regulation of enzymes responsible for the endocannabinoids metabolism, by the inhibition of reactive oxygen species release and the modulation of JNK/p38 cascade with consequent NF-κB p65 nuclear translocation suppression. Our data suggest that C. sativa phytocomplex and its multitarget mechanism could represent a novel therapeutic strategy for neuroinflammatory-related diseases.

OBJECTIVES The exposure of neurons to an excessive excitatory stimulation induces the alteration of the normal neuronal function. Mood disorders are among the first signs of alterations in the central nervous system function. Magnolia officinalis bark extract has been extensively used in the traditional medicine systems of several countries, showing several pharmacological activities. Honokiol, the main constituent of M. officinalis, is a GABA modulator and a CB1 agonist, which is deeply investigated for its role in modulating mood disorders. METHODS Thus, we evaluated the possible neuroprotective effect of a standardized M. officinalis bark extract (MOE), enriched in honokiol, and its effect on animal mood behavioural tests and in an in vitro model of excitotoxicity. KEY FINDINGS MOE showed neuroprotective effect using SH-SY5Y cells, by normalizing brain-derived neurotrophic factor release. Then, we tested the effect of MOE in different behavioural tests evaluating anxiety and depression and we observed a selective anxiolytic-like effect. Finally, we confirmed the involvement of CB1 in the final effect of MOE by the co-administration of the CB1 antagonist, AM251. CONCLUSION These results suggest that MOE could be considered an effective and safe anxiolytic candidate with neuroprotective activity.

Background: The importance of polyphenols in human health is well known; these compounds are common in foods, such as fruits, vegetables, spices, extra virgin olive oil and wine. On the other hand, the different factors that modulate the biological activity of these compounds are less well known. Conceptualization of the work: In this review we took into account about 200 relevant and recent papers on the following topics: “polyphenols bioavailability”, “polyphenols matrix effect”, “food matrix effect”, “polyphenols-cytochromes interaction”, after having reviewed and updated information on chemical classification and main biological properties of polyphenols, such as the antioxidant, anti-radical and anti-inflammatory activity, together with the tricky link between in vitro tests and clinical trials. Key findings: the issue of polyphenols bioavailability and matrix effect should be better taken into account when health claims are referred to polyphenols, thus considering the matrix effect, enzymatic interactions, reactions with other foods or genetic or gender characteristics that could interfere. We also discovered that in vitro studies often underrate the role of phytocomplexes and thus we provided practical hints to describe a clearer way to approach an investigation on polyphenols for a more resounding transfer to their use in medicine.

This study aimed to characterize the chemical profile of an ethanolic extract of Tuscan Rosmarinus officinalis (Roex) and to determine its in vitro bioactivity. The content of phenolic and flavonoid compounds, hydroxycinnamic acids and triterpenoids was determined, and high-performance liquid chromatography-diode array detection (HPLC-DAD) analysis revealed that rosmarinic acid and other hydroxycinnamic derivatives were the main constituents of the extract. Roex demonstrated to have both antioxidant activity and the capability to scavenge hydrogen peroxide in a concentration dependent manner. Moreover, NIH3T3 mouse fibroblasts and human breast adenocarcinoma cells MDA-MB-231 viability was influenced by the extract with an IC50 of 2.4 × 10−1 mg/mL and 4.8 × 10−1 mg/mL, respectively. The addition of Roex to the culture medium of both the above cell lines, resulted also in the reduction of cell death after H2O2 pre-treatment. The Ames test demonstrated that Roex was not genotoxic towards both TA98 and TA100 strains, with and without S9 metabolic activation. The extract, by inactivating thrombin, showed to also have an anti-coagulating effect at low concentration values. All these biological activities exerted by Roex are tightly correlated to its phytochemical profile, rich in bioactive compounds.