Metabolism and Effects of Dietary Phenolic Acids.

Ferulic acid is a secondary metabolite usually found as esters in plants and dihydrocaffeic acid results from the microbial metabolism of flavonoids and of caffeic acid, both widely distributed in food. Even though ferulic acid and flavonoids have been proposed to exert several beneficial effects on health, their in vivo activities could partly result from their microbial metabolites and strongly depend on their bioavailability. The absorption and metabolism of phenolic acids were studied in vitro with a model for the colonic epithelium composed of absorptive and mucus secreting cells, ex vivo with everted colonic sacs and liver slices, and in vivo with rats. The photoprotective effect of phenolic acids was tested in vitro on the keratinocytes HaCaT. The ferulic acid permeation was mainly by transcellular diffusion and also by a facilitated transport (S-MCT and MCT1). Intestinal cells conjugated ferulic acid with sulphate or glucuronide and reduced its unsaturated side chain. In rats, intestinal cells were more potent for glucuronidation of dihydrocaffeic acid, whereas the liver favoured sulphation, the methylation being regio-selective. Intestinal and hepatic cells oxidized dihydroferulic acid into ferulic acid, which they reduced into dihydroferulic acid. HaCaT cells were able of sulphation, methylation and reduction. Dihydrocaffeic acid decreased the cytotoxicity and the production of IL-6 and IL-8 in HaCaT cells following UV radiation, the minimum structure required for such effect consisting in a propionic side chain attached to a phenyl ring substituted with a catechol moiety. Dihydrocaffeic acid also protected erythrocytes from lysis induced by a free radical initiator. Phenolic acids released from food by the colonic microflora can be conjugated and metabolized by the intestinal epithelium, liver and other tissues, such as the epidermis, strongly reducing the circulating amount of the parent compound, which could be the most active form

Metabolism and Effects of Dietary Phenolic Acids.

Ferulic acid is a secondary metabolite usually found as esters in plants and dihydrocaffeic acid results from the microbial metabolism of flavonoids and of caffeic acid, both widely distributed in food. Even though ferulic acid and flavonoids have been proposed to exert several beneficial effects on health, their in vivo activities could partly result from their microbial metabolites and strongly depend on their bioavailability. The absorption and metabolism of phenolic acids were studied in vitro with a model for the colonic epithelium composed of absorptive and mucus secreting cells, ex vivo with everted colonic sacs and liver slices, and in vivo with rats. The photoprotective effect of phenolic acids was tested in vitro on the keratinocytes HaCaT. The ferulic acid permeation was mainly by transcellular diffusion and also by a facilitated transport (S-MCT and MCT1). Intestinal cells conjugated ferulic acid with sulphate or glucuronide and reduced its unsaturated side chain. In rats, intestinal cells were more potent for glucuronidation of dihydrocaffeic acid, whereas the liver favoured sulphation, the methylation being regio-selective. Intestinal and hepatic cells oxidized dihydroferulic acid into ferulic acid, which they reduced into dihydroferulic acid. HaCaT cells were able of sulphation, methylation and reduction. Dihydrocaffeic acid decreased the cytotoxicity and the production of IL-6 and IL-8 in HaCaT cells following UV radiation, the minimum structure required for such effect consisting in a propionic side chain attached to a phenyl ring substituted with a catechol moiety. Dihydrocaffeic acid also protected erythrocytes from lysis induced by a free radical initiator. Phenolic acids released from food by the colonic microflora can be conjugated and metabolized by the intestinal epithelium, liver and other tissues, such as the epidermis, strongly reducing the circulating amount of the parent compound, which could be the most active form

Predicting Phenolic Acid Absorption in Caco-2 Cells: A Theoretical Permeability Model and Mechanistic Study

There is a considerable need to rationalize the membrane permeability and mechanism of transport for potential nutraceuticals. The aim of this investigation was to develop a theoretical permeability equation, based on a reported descriptive absorption model, enabling calculation of the transcellular component of absorption across Caco-2 monolayers. Published data for Caco-2 permeability of 30 drugs transported by the transcellular route were correlated with the descriptors 1-octanol/water distribution coefficient (log D, pH 7.4) and size, based on molecular mass. Nonlinear regression analysis was used to derive a set of model parameters a?, ??, and b? with an integrated molecular mass function. The new theoretical transcellular permeability (TTP) model obtained a good fit of the published data (R2 = 0.93) and predicted reasonably well (R2 = 0.86) the experimental apparent permeability coefficient (Papp) for nine non-training set compounds reportedly transported by the transcellular route. For the first time, the TTP model was used to predict the absorption characteristics of six phenolic acids, and this original investigation was supported by in vitro Caco-2 cell mechanistic studies, which suggested that deviation of the Papp value from the predicted transcellular permeability (Papptrans) may be attributed to involvement of active uptake, efflux transporters, or paracellular flux

Absorption and Metabolism of Chlorogenic Acids in Cultured Gastric Epithelial Monolayers

Gastric absorption of feruloylquinic acid and di-O-caffeoylquinic acid analogs has never been investigated despite their potential contribution to the proposed beneficial health effects leading to reduced risk of type 2 diabetes. Using a cultured gastric epithelial model, with an acidic apical pH, the relative permeability coefficients (Papp) and metabolic fate of a series of chlorogenic acids (CGAs) were investigated. Mechanistic studies were performed in the apical to basal direction and demonstrated differential rates of absorption for different CGA subgroups. For the first time, we show intact absorption of feruloylquinic acids and caffeoylquinic acid lactones across the gastric epithelium (Papp ∼ 0.2 cm/s). Transport seemed to be mainly by passive diffusion, because good linearity was observed over the incubation period and test concentrations, and we speculate that a potential carrier-mediated component may be involved in uptake of certain 4-acyl CGA isomers. In contrast, absorption of intact di-O-caffeoylquinic acids was rapid (Papp ∼ 2–10 cm/s) but nonlinear with respect to time and concentration dependence, which was potentially limited by interaction with an efflux transporter and/or pH gradient dependence. For the first time, methylation is shown in gastric mucosa. Furthermore, isoferulic acid, dimethoxycinnamic acid, and ferulic acid were identified as novel gastric metabolites of CGA biotransformation. We propose that the stomach is the first location for the release of hydroxycinnamic acids, which could explain their early detection after coffee consumption

Circulating Structurally Related (-)-Epicatechin Metabolite Species and Levels after Sustained Intake of a Cocoa Powder High in Polyphenols Are Comparable to Those Achieved after a Single Dose

Background: While the bioavailability of cocoa polyphenols, particularly of the monomer (-)-epicatechin, has been investigated after a single-dose intake, the effect of sustained cocoa consumption on the metabolic profile of the structurally related (-)-epicatechin metabolites (SREMs) has not been investigated. Methods: A randomized, controlled crossover clinical trial in healthy young adults (18–40 year) was conducted to evaluate SREMs after consumption of a single-dose and after daily consumption of 1.3 g of polyphenol-rich cocoa powder for 28 days. The circulating SREMs were measured by ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Results: Twenty subjects (eleven males and nine females) were enrolled. The SREMs concentrations increased to 1741 ± 337 nM after a single-dose and to 1445 ± 270 nM after sustained supplementation. Sulfate conjugates showed higher levels in females (p < 0.05). The epicatechin-3′-glucuronide (E3′G) and epicatechin-3′-sulfate (E3′S) were the most abundant metabolites in all subjects. A high intra-individual correlation (r = 0.72, p < 0.001) between SREMs concentrations after single-dose and sustained supplementation was observed. The antioxidant capacity of plasma did not change in response to the intervention and was not correlated with any of the SREMs. Conclusion: The individual SREMs profile and concentrations after a 28-day supplementation are comparable to those after a single dose

Clostridium difficile Biofilm: Remodeling Metabolism and Cell Surface to Build a Sparse and Heterogeneously Aggregated Architecture

International audienceClostridium difficile is an opportunistic entero-pathogen causing post-antibiotic and nosocomial diarrhea upon microbiota dysbiosis. Although biofilms could contribute to colonization, little is known about their development and physiology. Strain 630Δerm is able to form, in continuous-flow micro-fermentors, macro-colonies and submersed biofilms loosely adhesive to glass. According to gene expression data, in biofilm/planktonic cells, central metabolism is active and fuels fatty acid biosynthesis rather than fermentations. Consistently, succinate is consumed and butyrate production is reduced. Toxin A expression, which is coordinated to metabolism, is down-regulated, while surface proteins, like adhesins and the primary Type IV pili subunits, are over-expressed. C-di-GMP level is probably tightly controlled through the expression of both diguanylate cyclase-encoding genes, like dccA, and phosphodiesterase-encoding genes. The coordinated expression of genes controlled by c-di-GMP and encoding the putative surface adhesin CD2831 and the major Type IV pilin PilA1, suggests that c-di-GMP could be high in biofilm cells. A Bacillus subtilis SinR-like regulator, CD2214, and/or CD2215, another regulator co-encoded in the same operon as CD2214, control many genes differentially expressed in biofilm, and in particular dccA, CD2831 and pilA1 in a positive way. After growth in micro-titer plates and disruption, the biofilm is composed of robust aggregated structures where cells are embedded into a polymorphic material. The intact biofilm observed in situ displays a sparse, heterogeneous and high 3D architecture made of rods and micro-aggregates. The biofilm is denser in a mutant of both CD2214 and CD2215 genes, but it is not affected by the inactivation of neither CD2831 nor pilA1 . dccA, when over-expressed, not only increases the biofilm but also triggers its architecture to become homogeneous and highly aggregated, in a way independent of CD2831 and barely dependent of pilA1 . Cell micro-aggregation is shown to play a major role in biofilm formation and architecture. This thorough analysis of gene expression reprogramming and architecture remodeling in biofilm lays the foundation for a deeper understanding of this lifestyle and could lead to novel strategies to limit C. difficile spread

Intragenic FMR1 disease-causing variants: a significant mutational mechanism leading to Fragile-X syndrome

International audienceFragile-X syndrome (FXS) is a frequent genetic form of intellectual disability (ID). The main recurrent mutagenic mechanism causing FXS is the expansion of a CGG repeat sequence in the 5'-UTR of the FMR1 gene, therefore, routinely tested in ID patients. We report here three FMR1 intragenic pathogenic variants not affecting this sequence, identified using high-throughput sequencing (HTS): a previously reported hemizygous deletion encompassing the last exon of FMR1, too small to be detected by array-CGH and inducing decreased expression of a truncated form of FMRP protein, in three brothers with ID (family 1) and two splice variants in boys with sporadic ID: a de novo variant c.990+1G > A (family 2) and a maternally inherited c.420-8A > G variant (family 3). After clinical reevaluation, the five patients presented features consistent with FXS (mean Hagerman's scores=15). We conducted a systematic review of all rare non-synonymous variants previously reported in FMR1 in ID patients and showed that six of them are convincing pathogenic variants. This study suggests that intragenic FMR1 variants, although much less frequent than CGG expansions, are a significant mutational mechanism leading to FXS and demonstrates the interest of HTS approaches to detect them in ID patients with a negative standard work-up