Circulating human serum metabolites derived from the intake of a saffron extract (Safr’Inside™) protect neurons from oxidative stress: Consideration for depressive disorders

Increases in oxidative stress have been reported to play a central role in the vulnerability to depression, and antidepressant drugs may reduce increased oxidative stress in patients. Among the plants exerting anti-inflammatory and anti-oxidant properties, saffron, a spice derived from the flower of Crocus sativus, is also known for its positive effects on depression, potentially through its SSRI-like properties. However, the molecular mechanisms underlying these effects and their health benefits for humans are currently unclear. Using an original ex vivo clinical approach, we demonstrated for the first time that the circulating human metabolites produced following saffron intake (Safr’Inside™ ) protect human neurons from oxidative-stress-induced neurotoxicity by preserving cell viability and increasing BNDF production. In particular, the metabolites significantly stimulated both dopamine and serotonin release. In addition, the saffron’s metabolites were also able to protect serotonergic tone by inhibiting the expression of the serotonin transporter SERT and down-regulating serotonin metabolism. Altogether, these data provide new biochemical insights into the mechanisms underlying the beneficial impact of saffron on neuronal viability and activity in humans, in the context of oxidative stress related to depression

Benefits of Circulating Human Metabolites from Fish Cartilage Hydrolysate on Primary Human Dermal Fibroblasts, an Ex Vivo Clinical Investigation for Skin Health Applications

International audienceDue to its significant exposure to stressful environmental factors, the skin undergoes a high remodeling rate over time, which alters not only its appearance but also its functionality. This alteration of the skin, namely photoaging, is characterized by dryness and a loss of elasticity that mainly originates from the dysregulation of dermal fibroblast activities. In order to overcome such tissue outcome, cosmetic products have evolved toward nutricosmetics, thus promoting beauty from within. Among bio-actives of interest, bio-peptides deriving from plant or animal sources may exert various biological activities beyond their nutritional value. However, studies remain mostly descriptive and the mode of action at the cellular level in clinic remains a concern. In a recent clinical trial, it was showed that supplementation with a fish cartilage hydrolysate (FCH) improved signs of chronological and photoaging-induced skin changes in healthy women. Here, using an original ex vivo clinical approach adapted to nutricosmetic purpose, we further demonstrated that this fish cartilage hydrolysate was absorbed and that the circulating metabolites produced in humans following FCH intake stimulate human dermal fibroblast growth, promote specific hyaluronan production, up-regulate elastin synthesis and inhibit MMP-1 and 3 expression along with the enhancement of TGF-β release. Altogether, these data provide clues on the mechanisms likely contributing to the beneficial impact of FCH on human skin functionality by supporting hydration, elasticity and limiting the expression of catabolic factors involved in photoaging onset

Human Serum, Following Absorption of Fish Cartilage Hydrolysate, Promotes Dermal Fibroblast Healing through Anti-Inflammatory and Immunomodulatory Proteins

International audienceBackground/Objectives: Marine collagen peptides (MCPs) and glycosaminoglycans (GAGs) have been described as potential wound-healing (WH) agents. Fish cartilage hydrolysate (FCH) is a natural active food ingredient obtained from enzymatic hydrolysis which combines MCPs and GAGs. Recently, the clinical benefits of FCH supplementation for the skin, as well as its mode of action, have been demonstrated. Some of the highlighted mechanisms are common to the WH process. The aim of the study is therefore to investigate the influence of FCH supplementation on the skin healing processes and the underlying mechanisms. Methods: To this end, an ex vivo clinical approach, which takes into account the clinical digestive course of nutrients, coupled with primary cell culture on human dermal fibroblasts (HDFs) and ultra-deep proteomic analysis, was performed. The effects of human serum enriched in circulating metabolites resulting from FCH ingestion (FCH-enriched serum) were assessed on HDF WH via an in vitro scratch wound assay and on the HDF proteome via diaPASEF (Data Independent Acquisition—Parallel Accumulation Serial Fragmentation) proteomic analysis. Results: Results showed that FCH-enriched human serum accelerated wound closure. In support, proteins with anti-inflammatory and immunomodulatory properties and proteins prone to promote hydration and ECM stability showed increased expression in HDFs after exposure to FCH-enriched serum. Conclusions: Taken together, these data provide valuable new insights into the mechanisms that may contribute to FCH’s beneficial impact on human skin functionality by supporting WH. Further studies are needed to reinforce these preliminary data and investigate the anti-inflammatory and immunomodulatory properties of FCH

Reduced Production of Pro-Inflammatory and Pro-Catabolic Factors by Human Serum Metabolites Derived from a Patented Saffron Extract Intake

Safe and anti-inflammatory plant-based natural products present an increasing focus in the treatment of chronic inflammatory diseases such as osteoarthritis or inflammatory bowel diseases. Among them, saffron, a spice derived from the stigma of Crocus sativus, could have anti-inflammatory properties and would be therefore a promising therapeutic agent for the treatment of such conditions. However, the anti-inflammatory molecular mechanisms of saffron in humans are still understudied and unclear. In this study, combining human serum metabolites and cell cultures, we evaluated the effect of circulating metabolites from the consumption of a patented saffron extract (Safr’InsideTM) on the chondrocytes and colon epithelial cell responses to inflammatory stress. Parametric or non-parametric Analysis of Variance with post hoc tests was performed. We demonstrated that human serum containing metabolites from saffron intake attenuated IL-1β-stimulated production of PGE2 and MMP-13 in chondrocyte cells and limited the increase in ICAM-1, MCP-1, iNOS, and MMP-3 in human epithelial cells following combined IL-1β and TNF-α inflammatory stimulation. Altogether, these data provide new findings into the mechanisms underlying the beneficial effects of saffron on chondrocytes and enterocyte cells at the cellular level and in the context of chronic inflammatory disorders

Metabolic and Anti-Inflammatory Protective Properties of Human Enriched Serum Following Artichoke Leaf Extract Absorption: Results from an Innovative Ex Vivo Clinical Trial

International audienceThe aging of our population is accompanied by an increased prevalence of chronic diseases. Among those, liver, joint and adipose tissue-related pathologies have a major socio-economic impact. They share common origins as they result from a dysregulation of the inflammatory and metabolic status. Plant-derived nutrients and especially polyphenols, exert a large range of beneficial effects in the prevention of chronic diseases but require clinically validated approaches for optimized care management. In this study, we designed an innovative clinical approach considering the metabolites produced by the digestive tract following the ingestion of an artichoke leaf extract. Human serum, enriched with metabolites deriving from the extract, was collected and incubated with human hepatocytes, human primary chondrocytes and adipocytes to determine the biological activity of the extract. Changes in cellular behavior demonstrated that the artichoke leaf extract protects hepatocytes from lipotoxic stress, prevents adipocytes differentiation and hyperplasia, and exerts chondroprotective properties in an inflammatory context. These data validate the beneficial health properties of an artichoke leaf extract at the clinical level and provide both insights and further evidence that plant-derived nutrients and especially polyphenols from artichoke may represent a relevant alternative for nutritional strategies addressing chronic disease issues

Metabolic profiling strategy for determination of the absorption peak of artichoke leaf extract in human enriched serum

National audienceOur society nowadays is aging and the chronic disease appearance is increasing. Toprevent this, plant-derived nutrients like polyphenols could be used. They are known toprotect tissues from inflammatory and metabolic dysregulation induced by chronic diseases.Polyphenols are found in high level in artichoke leaf extract (ALE) and his ingestion may beeffective to prevent such dysregulations. To prove this, an innovative clinical approach wasdesigned. Patients ingested ALE and their blood was collected every 20 min after theingestion during 240 min. The kinetic profile of the ALE absorption was studied by1H NMRspectroscopy. The comparison of 1D spectra showed that the highest variations wereobserved in the phenolic compounds’ region and for the 100 min samples. The addition ofthe degradation products of chlorogenic acid, the major ALE polyphenol compound, to anaive serum led to similar chemical shift variations to those observed for the 100 minsamples suggesting that the metabolic changes in the serum profile after ingestion were dueto the presence of the ALE polyphenol metabolites. These results, then, allowed to continuethe clinical trial and to determine the influence of sera enriched in metabolites resultingfrom ALE consumption on the behavior of human hepatocyte, adipocyte and chondrocytecultures. Altogether, the obtained data show that polyphenols from artichoke may representa relevant alternative for nutritional strategies addressing chronic disease issue [1].References:1. Wauquier F.; Boutin-Wittrant L.; Viret A.; Guilhaudis L et al. Nutrients. 2021, 13 (8), 2653.DOI: https://doi.org/10.3390/nu1308265

Oral supplementation with fish cartilage hydrolysate in an adult population suffering from knee pain and function discomfort: results from an innovative approach combining an exploratory clinical study and an ex vivo clinical investigation

Abstract Background Aging is frequently associated with impairments of the musculoskeletal system and many elderly people experience joint discomfort or pain which might reduce their ability to move and consequently alter their quality of life. A beneficial effect of fish cartilage hydrolysate (FCH) on pain and joint function has recently been shown in an ACLT/pMMx osteoarthritis rat model. Methods We therefore performed an exploratory, non-comparative, multi-centric clinical trial including 33 subjects with moderate knee joint discomfort and loss of functionality to investigate the efficacy of FCH on their algo-functional status. We further determined the potential health benefit of FCH in an original clinical ex vivo study investigating the role of FCH human metabolites on primary human chondrocytes. Results FCH significantly improved knee pain and function, as assessed by the Knee injury and Osteoarthritis Outcome Score (KOOS). Moreover, FCH significantly reduced pain at rest and while walking, and patient global assessment (PGA), as assessed by the Visual Analogue Scale (VAS), and improved patients’ quality of life (SF-36). FCH metabolites decreased the synthesis of catabolic factors (MMP-13) and pro-inflammatory mediators (NO, PGE2) and limited the inhibitory effect of IL-1β on the synthesis of cartilage matrix components (GAG and collagen). Conclusions Thus, these data provide insights on the mode of action of FCH in humans and contribute to explain how FCH may relieve pain and improve joint function in subjects with knee discomfort. Although these preliminary data need to be confirmed in a randomized controlled trial, they strongly support the potential health benefit of such an active ingredient. Trial registration: The study was registered on clinicaltrials.gov with the identifier NCT04420091 (09/06/2020)

Circulating Human Metabolites Resulting From TOTUM-070 Absorption (a Plant-Based, Polyphenol-Rich Hypocholesterolemic Ingredient) Improve Lipid Metabolism in Human Hepatocytes: Lessons From an Original ex vivo Clinical Trial

Introduction: TOTUM-070 is a patented polyphenol-rich compound which has shown hypocholesterolemic properties in preclinical studies. However, clinically validated approaches and further investigations on the mode of action at cellular level especially in humans are required for optimized care management. In this study, we designed an ex-vivo clinical innovative approach considering the metabolites produced by the digestive tract following the ingestion of TOTUM-070 in humans.Methods: Human serum was collected in healthy subjects before and following acute intake of 5g of TOTUM-070. Availability of circulating metabolites was confirmed and characterized by UPLC-MS. Human serum enriched with metabolites deriving from TOTUM-070 absorption was further incubated with human hepatocytes, pretreated or not with palmitate (250 μM). In such lipotoxic environment, hepatocyte behavior was monitored to determine whether and how TOTUM-070 metabolites may improve cholesterol metabolism in human.Results: In the presence of the human metabolites from TOTUM-070, human hepatocytes were protected from an induced lipotoxic stress. No effect on cell toxicity was detected in the presence of enriched sera. Hepatocyte protection was characterized by (1) the inhibition of both triglycerides (-41%, p<0.001) and cholesterol (-50%, p<0.001) storage, (2) a reduced de novo cholesterol synthesis (HMG-CoA reductase activity reduced by 44%, p<0.001), (3) a lowered fatty acid synthase expression (p<0.001), (4) the stimulation of the Lecithin Cholesterol Acyl Transferase (LCAT) activity (p<0.05) and finally (5) a higher CYP7A1 expression (p<0.05). All together, these data provide new biochemical insights into the mechanisms underlying, in humans, the beneficial impact of TOTUM-070 on lipid metabolism in liver cells.Conclusion: Using a pioneering clinical ex vivo approach considering the digestive processes of nutrients, we give clues on the role of circulating metabolites produced following TOTUM-070 intake in humans in hepatocyte protection

Metabolic profiling strategy for determination of the absorption peak of artichoke leaf extract in human enriched serum

National audienceOur society nowadays is aging and the chronic disease appearance is increasing. Toprevent this, plant-derived nutrients like polyphenols could be used. They are known toprotect tissues from inflammatory and metabolic dysregulation induced by chronic diseases.Polyphenols are found in high level in artichoke leaf extract (ALE) and his ingestion may beeffective to prevent such dysregulations. To prove this, an innovative clinical approach wasdesigned. Patients ingested ALE and their blood was collected every 20 min after theingestion during 240 min. The kinetic profile of the ALE absorption was studied by1H NMRspectroscopy. The comparison of 1D spectra showed that the highest variations wereobserved in the phenolic compounds’ region and for the 100 min samples. The addition ofthe degradation products of chlorogenic acid, the major ALE polyphenol compound, to anaive serum led to similar chemical shift variations to those observed for the 100 minsamples suggesting that the metabolic changes in the serum profile after ingestion were dueto the presence of the ALE polyphenol metabolites. These results, then, allowed to continuethe clinical trial and to determine the influence of sera enriched in metabolites resultingfrom ALE consumption on the behavior of human hepatocyte, adipocyte and chondrocytecultures. Altogether, the obtained data show that polyphenols from artichoke may representa relevant alternative for nutritional strategies addressing chronic disease issue [1].References:1. Wauquier F.; Boutin-Wittrant L.; Viret A.; Guilhaudis L et al. Nutrients. 2021, 13 (8), 2653.DOI: https://doi.org/10.3390/nu1308265

Les métabolites sériques humains circulants dérivés de la consommation d'un extrait de safran (Safr'InsideTM) protègent les neurones du stress oxydatif : considérations pour les troubles dépressifs

International audienceIntroduction et but de l’étude : Il a été démontré que l’augmentation du stress oxydatif joue un rôle central dans la vulnérabilité à la dépression, et les antidépresseurs peuvent réduire l’augmentation du stress oxydatif chez les patients. Parmi les plantes aux propriétés anti-inflammatoires et antioxydantes, le safran, une épice dérivée de la fleur de Crocus sativus, est également connu pour ses effets positifs sur la dépression, potentiellement grâce à ses propriétés de recapture de la sérotonine. Cependant, les mécanismes moléculaires sous-jacents à ces effets et leurs bienfaits pour la santé humaine ne sont pas encore clairs.Matériel et méthodes : En utilisant une approche clinique ex vivo originale, nous avons démontré pour la première fois que les métabolites humains circulants et produits suite à la prise de safran (Safr’InsideTM) protègent les neurones humains de la neurotoxicité induite par le stress oxydatif.Un groupe de 10 hommes sains (âge : 25,0 ans, +/-5,1 ; IMC : 23,9 kg/m2, +/-2,3 ; &gt;60 kg ; sans traitement médicamenteux ; et sans distinction d’origine ethnique) se sont portés volontaires pour cette étude. La première étape de l’étude visait à déterminer le pic d’absorption des métabolites de l’extrait de safran, et notamment des crocétines. Dix volontaires sains à jeun depuis 12 h ont reçu 300 mg d’un extrait de safran (SaE). Une fois le pic d’absorption déterminé, les volontaires ont été rappelés pour le prélèvement de la fraction sérique enrichie. Pour cette deuxième phase clinique, huit volontaires sains à jeun depuis 12 h ont reçu 300 mg de SaE sous forme d’une capsule. 50 mL de sang veineux ont été prélevés avant et après 'ingestion pour le prélèvement d'un sérum naïf et enrichi. Ces sérums ont ensuite été rendus compatibles avec la culture de cellules humaines selon une méthodologie protégée et exploitée par Clinic’n’Cell SAS. Des neurones humains ont été incubés avec ces sérums pour déterminer l’influence des métabolites de l’extrait sur le comportement cellulaire.Pour analyser les effets des métabolites sériques de SaE sur le stress neurotoxique, des cellules SH-SY5Y différenciées ont été préincubées pendant 24h dans le milieu MEM/F12 (1:1) en présence de 10 % de sérum naïf humain ou de sérum enrichi humain, selon le protocole Clinic’n’Cell avant un traitement supplémentaire de 24 heures avec 500 μM H2O2.Résultats et analyses statistiques: Pour suivre l’enrichissement des métabolites dans le sérum, nous avons utilisé la crocétine comme marqueur précis et validé de l’absorption de SaE. Les volontaires à jeun ont reçu 300 mg de SaE et le profil d’absorption des crocétines a été surveillé pendant 5 h. L’analyse du profil d’absorption a montré que la concentration maximale de crocétine était atteinte 90 min après l’ingestion (données non présentées). Par conséquent, du sérum enrichi en métabolites SaE a été collecté 90 min après l’ingestion pour la deuxième phase du protocole clinique.Ex vivo, nous avons démontré pour la première fois que les métabolites humains circulants produits suite à la prise de safran (Safr’InsideTM) protègent les neurones humains d'une neurotoxicité induite par un stress oxydatif en préservant la viabilité cellulaire et en augmentant la production de BNDF (+80%). En particulier, les métabolites ont stimulé de manière significative la libération de dopamine (+184%) et de sérotonine (+37%). De plus, les métabolites du safran étaient également capables de protéger le tonus sérotoninergique en inhibant l’expression du transporteur de sérotonine SERT et en régulant à la baisse le métabolisme de la sérotonine (-51,9% 5-HIAA).Conclusion: Dans l’ensemble, ces données fournissent de nouvelles perspectives biochimiques sur les mécanismes sous-jacents au bénéfice du safran sur la viabilité et l’activité neuronale chez l’humain dans un contexte de stress oxydatif impliqué dans la dépression