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

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

Circulating bioactives resulting from totum-854 absorption in humans protects primary human endothelial cells against lipotoxicity: lessons from an original ex vivo clinical trial

International audienc

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

TOTUM-070 is a patented polyphenol-rich blend of five different plant extracts showing separately a latent effect on lipid metabolism and potential synergistic properties. In this study, we investigated the health benefit of such a formula. Using a preclinical model of high fat diet, TOTUM-070 (3 g/kg of body weight) limited the HFD-induced hyperlipemia with a reduction in triglyceride (−32% after 6 weeks; −20.3% after 12 weeks) and non-HDL cholesterol levels (−21% after 6 weeks; −38.4% after 12 weeks). To further investigate such a benefit and its underlying mechanisms in humans, we designed an ex vivo clinical approach to collect the circulating bioactives resulting from TOTUM-070 ingestion and to determine their biological activities on human hepatocytes. Human serum was obtained from healthy subjects before and after intake of TOTUM-070 (4995 mg). The presence of circulating metabolites was assessed by UPLC-MS/MS. Serum containing metabolites was further incubated with hepatocytes cultured in a lipotoxic environment (palmitate, 250 µM). RNA sequencing analyses show that lipid metabolism was one of the most impacted processes. Using histologic, proteomic, and enzymatic assays, the effects of human TOTUM-070 bioactives on hepatocyte metabolism were characterized by (1) the inhibition of lipid storage, including both (2) triglycerides (−41%, p p p p < 0.001). Altogether, these data support the beneficial impact of TOTUM-070 on lipid metabolism and provide new biochemical insights in human mechanisms occurring in liver cells

Additional file 1 of 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

Figure S1. Mean evolution (± SE) of the KOOS sub-scores over time: (a) KOOS Pain score over time; (b) KOOS Symptoms score over time; (c) KOOS Daily living activities score over time; (d) KOOS Sport and recreation function score over time; (e) Quality of life score over time ? FAS population. ***p ≤ 0.001; **p ≤ 0.01; *p ≤ 0.05; ns, not significant. Figure S2. Mean evolution (± SE) of the SF-36 sub-scores over time: (a) SF-36 Physical functioning over time; (b) SF-36 Role limitations due to physical health over time; (c) SF-36 Energy/Fatigue over time; (d) SF-36 Pain over time; (e) SF-36 General health over time ? FAS population. ***p ≤ 0.001; **p ≤ 0.01; *p ≤ 0.05; ns, not significant. Table S1. Subjects? knee discomfort history description at the inclusion in the study ? FAS population. Table S2. Results of the repeated measures ANOVA models for KOOS global score and each subscale, and mean difference and effect size between baseline and 3 months of follow-up ? PP population ? N=24. Table S3. Results of the repeated measures ANOVA models for SF-36 global score and each subscale ? PP population ? N=24. Table S4. Results of the repeated measures ANOVA models for knee pain at rest and while walking and PGA using VAS scale, and mean difference and effect size between baseline and 3 months of follow-up ? PP population ? N=24. Table S5. Comparison of patients? treatment response between the first and the second follow-up ? PP population ? N=24. Table S6. Results of the comparison between the first and the second follow-up for compliance? FAS population. Table S7. Comparisons of subjects satisfaction between the two follow-up visits ? FAS population. Table S8. Results of the analysis of the time evolution for pain killer use and frequency of intake ? FAS population. Table S9: Listing of AE and SAE by decreasing order of frequency ? Safety population ? N=28 adverse events. Table S10. Distribution of link with FCH and action taken in response to AE ? Safety population ? N=28 adverse events. Table S11. Distribution of subjects at each visit ? Safety population ? N=32 patients. Table S12. Complete list of exclusion criteria. Figure S3. Primary human chondrocytes subjected to ex vivo procedures for validation of cell viability in human serum. Cell viability was measured with an XTT-based assay upon either FCS or human serum incubation (H-NAIVE for human naive serum and H-FCH for human serum enriched with circulating FCH metabolites) for 24 h and 48 h (A and B). Measures were performed in quadruplicates per condition/volunteer (n=10 volunteers). Values are presented as mean ± SD. The differences were considered statistically significant at p < 0.05 with ** for p < 0.01 and **** for p < 0.0001