Binding Interaction Between Lauric Acid and Cluster of Differentiation 36 Underpinned by a Fluorescence- Intensifying Assay.

Cluster of differentiation 36 (CD36) is a scavenger receptor expressed in various vertebrate cells that contains diverse ligands, including long-chain fatty acids. This receptor has recently been suggested as a captor of specific volatile odorants (e.g., aliphatic acetates) in the mammalian nasal epithelium. This study used a fluorescence-intensifying assay to produce the first evidence that lauric acid, an odorous fatty acid, directly binds to CD36. This expansion of the repertoire of volatile ligands supports potential applications for nasal CD36. Our present findings could promote future research aimed at understanding the mechanisms of fatty acid interactions with CD36.

Striped catfish (Pangasianodon hypophthalmus) exploit food sources across anaerobic decomposition- and primary photosynthetic production-based food chains.

Dietary information from aquatic organisms is instrumental in predicting biological interactions and understanding ecosystem functionality. In freshwater habitats, generalist fish species can access a diverse array of food sources from multiple food chains. These may include primary photosynthetic production and detritus derived from both oxic and anoxic decomposition. However, the exploitation of anoxic decomposition products by fish remains insufficiently explored. This study examines feeding habits of striped catfish (Pangasianodon hypophthalmus) at both adult and juvenile stages within a tropical reservoir, using stable carbon, nitrogen, and sulfur isotope ratios (δ13C, δ15N, and δ34S, respectively) and fatty acid (FA) analyses. The adult catfish exhibited higher δ15N values compared to primary consumers that feed on primary photosynthetic producers, which suggests ingestion of food sources originating from primary photosynthetic production-based food chains. On the other hand, juvenile catfish demonstrated lower δ15N values than primary consumers, correlating with low δ34S value and large proportions of bacterial FA but contained small proportions of polyunsaturated FA. This implies that juveniles utilize food sources from both anoxic decomposition and primary photosynthetic production-based food chains. Our results indicate that food chains based on anoxic decomposition can indeed contribute to the dietary sources of tropical fish species.

Intestinal Absorption and Anti-Inflammatory Effects of Siphonein, a Siphonaxanthin Fatty Acid Ester from Green Algae.

Siphonein is a C19 acylated siphonaxanthin found in some edible green algae (e.g., Codium fragile and Caulerpa lentillifera). Although the content of siphonein in these green algae is similar to or higher than that of siphonaxanthin, studies of health-related biological activity of siphonein are much less than those of siphonaxanthin. Given the difference in the position of the acyl chain, one cannot infer intestinal absorption of siphonein from other general carotenoid fatty acid esters. In this study, we first investigated the intestinal absorption of siphonein using mouse and cell culture models. A small amount of siphonein was detected in the plasma of treated mice, and its concentration was higher than that of siphonaxanthin (i.e., the hydrolyzed product of ingested siphonein) from 1 to 6 h after administration. Pharmacological inhibition tests with differentiated Caco-2 cells showed that Nieman-Pick C1-like 1-mediated facilitated diffusion was involved in the cellular uptake of siphonein. These results indicate that, unlike general carotenoid fatty acid esters, siphonein can be absorbed without hydrolysis. We also evaluated the anti-inflammatory effect of siphonein in differentiated Caco-2 cells. Siphonein pretreatment modulated lipopolysaccharide-induced cellular lipidome alterations and suppressed mRNA expression of proinflammatory chemokines, CXCL8 protein release, and activation of NF-κB. This study provides new insights into the absorption processes of carotenoids and shows the anti-inflammatory effect of siphonein for the first time.

Gly-Pro protects normal human dermal fibroblasts from UVA-induced damages via MAPK-NF-κB signaling pathway.

Photoaging is characterized by skin dysfunction and wrinkle formation predominantly caused by chronic exposure to ultraviolet (UV) irradiation. Collagen peptides are well-recognized as nutritional supplements for enhancing skin health. Gly-Pro, a dipeptide found in collagen as a major repetitive sequence, is considered a prospect collagen peptide derivative that displays anti-photoaging potential. Herein, we evaluated the photoprotective effects of Gly-Pro in normal human dermal fibroblasts (NHDFs). Pretreatment by Gly-Pro at a concentration of 0.1 µM inhibited UVA-driven generation of reactive oxygen species (ROS) in NHDFs and attenuated UVA-induced changes in mRNA expression and activation of proteins of the MAPK-NF-κB signaling pathway. Meanwhile, Pro-Gly and cyclo(-Gly-Pro), two dipeptides that are structurally similar to Gly-Pro, depicted less anti-photoaging effects against UVA irradiation. Collectively, our data suggests that Gly-Pro has potential as a novel ingredient in nutricosmetic products for skincare and anti-photoaging.

Sphingolipids as Functional Food Components: Benefits in Skin Improvement and Disease Prevention.

Sphingolipids are ubiquitous components in eukaryotic organisms and have attracted attention as physiologically functional lipids. Sphingolipids with diverse structures are present in foodstuffs as these structures depend on the biological species they are derived from, such as mammals, plants, and fungi. The physiological functions of dietary sphingolipids, especially those that improve skin barrier function, have recently been noted. In addition, the roles of dietary sphingolipids in the prevention of diseases, including cancer and metabolic syndrome, have been studied. However, the mechanisms underlying the health-improving effects of dietary sphingolipids, especially their metabolic fates, have not been elucidated. Here, we review dietary sphingolipids, including their chemical structures and contents in foodstuff; digestion, intestinal absorption, and metabolism; and nutraceutical functions, based on the available evidence and hypotheses. Further research is warranted to clearly define how dietary sphingolipids can influence human health.

Integration of bioassay and non-target metabolite analysis of tomato reveals that ß-carotene and lycopene activate the adiponectin signaling pathway, including AMPK phosphorylation.

Adiponectin, an adipokine, regulates glucose metabolism and insulin sensitivity through the adiponectin receptor (AdipoR). In this study, we searched for metabolites that activate the adiponectin signaling pathway from tomato (Solanum lycopersicu). Metabolites of mature tomato were separated into 55 fractions by liquid chromatography, and then each fraction was examined using the phosphorylation assay of AMP-protein kinase (AMPK) in C2C12 myotubes and in AdipoR-knockdown cells by small interfering RNA (siRNA). Several fractions showed AMPK phosphorylation in C2C12 myotubes and siRNA-mediated abrogation of the effect. Non-targeted metabolite analysis revealed the presence of 721 diverse metabolites in tomato. By integrating the activity of fractions on AMPK phosphorylation and the 721 metabolites based on their retention times of liquid chromatography, we performed a comprehensive screen for metabolites that possess adiponectin-like activity. As the screening suggested that the active fractions contained four carotenoids, we further analyzed ß-carotene and lycopene, the major carotenoids of food. They induced AMPK phosphorylation via the AdipoR, Ca2+/calmodulin-dependent protein kinase kinase and Ca2+ influx, in addition to activating glucose uptake via AdipoR in C2C12 myotubes. All these events were characteristic adiponectin actions. These results indicated that the food-derived carotenoids, ß-carotene and lycopene, activate the adiponectin signaling pathway, including AMPK phosphorylation.

Early secretory pathway-resident Zn transporter proteins contribute to cellular sphingolipid metabolism through activation of sphingomyelin phosphodiesterase 1.

Sphingomyelin phosphodiesterase 1 (SMPD1) converts sphingomyelin into ceramide and phosphocholine; hence, loss of SMPD1 function causes abnormal accumulation of sphingomyelin in lysosomes, which results in the lipid-storage disorder Niemann-Pick disease (types A and B). SMPD1 activity is dependent on zinc, which is coordinated at the active site of the enzyme, and although SMPD1 has been suggested to acquire zinc at the sites where the enzyme is localized, precisely how SMPD1 acquires zinc remains to be clarified. Here, we addressed this using a gene-disruption/reexpression strategy. Our results revealed that Zn transporter 5 (ZNT5)-ZNT6 heterodimers and ZNT7 homodimers, which localize in the compartments of the early secretory pathway, play essential roles in SMPD1 activation. Both ZNT complexes contribute to cellular sphingolipid metabolism by activating SMPD1 because cells lacking the functions of the two complexes exhibited a reduced ceramide to sphingomyelin content ratio in terms of their dominant molecular species and an increase in the sphingomyelin content in terms of three minor species. Moreover, mutant cells contained multilamellar body-like structures, indicative of membrane stacking and accumulation, in the cytoplasm. These findings provide novel insights into the molecular mechanism underlying the activation of SMPD1, a key enzyme in sphingolipid metabolism.

Oral supplementation of sea cucumber and its hydrolysate mitigates ultraviolet A-induced photoaging in hairless mice.

BACKGROUND: Chronic exposure to ultraviolet (UV) radiation promotes skin photoaging, which is clinically characterized by dryness, laxity, and wrinkling. Sea cucumber (Stichopus japonicus) (SC) is a marine organism with culinary and medicinal applications, especially in Asian countries. It is also a potential nutraceutical as it exhibits bioactive effects, such as antioxidant, antitumor, and anticancer activity. This study examined the effects of SC and its hydrolysate (SCH) on ultraviolet A (UVA) induced skin barrier function and wrinkle formation using hairless mice. RESULTS: Ultraviolet A significantly induced transepidermal water loss and wrinkle formation, which were significantly mitigated upon oral administration of SC and SCH. Sea cucumber also mitigated the UVA-induced downregulation of epidermal natural moisturizing factors and the upregulation of Aqp3, Mmp13, Tnfa, and Il6 mRNA levels in the mouse skin. CONCLUSION: Taken together, these results suggest that dietary SC and SCH exert anti-photoaging effects by modulating filaggrin synthesis and desquamation in the epidermis and regulating the NF-κB pathway in the skin. Our research indicates that SC and SCH have potential applications in nutricosmetics for photoaging. © 2021 Society of Chemical Industry.

Multivariate Analysis Reveals That Unsubstituted ß-Ring and C8-Keto Structures Are Important Factors for Anti-Inflammatory Activity of Carotenoids.

Carotenoids are natural lipophilic pigments with substantial health benefits. Numerous studies have demonstrated the anti-inflammatory activities of carotenoids, especially toward lipopolysaccharide-induced inflammatory responses. As such, there are few reports on the evaluation and comparison of the anti-inflammatory activities of carotenoids against inflammation induced by other stimuli. In this study, we used pathogen-associated molecular patterns, proinflammatory cytokines, degenerated proteins, and chemical irritants as inflammatory inducers to evaluate the anti-inflammatory activities of eight different carotenoids. Each carotenoid showed characteristic anti-inflammatory activities; thus, we conducted a multivariate analysis to clarify the differences among them. Unsubstituted ß-ring (i.e., provitamin A) and C8-keto structures of carotenoids were found to be crucial for their inhibitory effects on the activation of nuclear factor-kappa B and interferon regulatory factors, respectively. Furthermore, we found that ß-carotene and echinenone treatment increased intracellular retinoid levels in monocytes and that the retinoids showed the similar activities to ß-carotene and echinenone. Taken together, the intake of both provitamin A and C8-keto carotenoids (e.g., siphonaxanthin and fucoxanthin) might be effective in improving the inflammatory status of individuals. A multivariate analysis of anti-inflammatory activities is a useful method for characterizing anti-inflammatory compounds.

Assessment of direct binding interaction between CD36 and its potential lipid ligands using a peptide mimic of the receptor labeled with a fluorophore.

Cluster of differentiation 36 (CD36) is a cell-surface receptor that recognizes diverse substances. We have presented indirect evidence that a short segment of the receptor comprising amino acids 149-168 contains a site for binding of its lipid ligands (e.g., distinct fatty acids and aldehydes). However, experimental support for their direct interactions is yet to be achieved. For this, we devised a fluorescence intensity assay, where a synthetic peptide consisting of CD36 amino acids 149-168 labeled with fluorescein isothiocyanate (FITC-CD36149-168) and its variant peptides were used as positive and negative probes, respectively. First, we obtained results indicating that 1-palmitoyl-2-(5-keto-6-octenedioyl)phosphatidylcholine (an established CD36 ligand) but not 1-palmitoyl-2-arachidonyl-phosphatidylcholine (a non-ligand of the receptor) bound in a saturable and specific manner to FITC-CD36149-168. Strikingly, the assay allowed us to provide the first evidence supporting direct and specific binding between the CD36 segment and fatty aldehydes (e.g., Z-11-hexadecenal). However, this method failed to illustrate specific interactions of the segment with fatty acids, such as oleic acid. Nonetheless, our findings offer further insight into the biologically relevant ligands and the role of CD36. In addition, we suggest that this fluorescence-based technique provides a convenient means to evaluate protein (peptide)-lipid interactions.

Dietary Ceramide Prepared from Soy Sauce Lees Improves Skin Barrier Function in Hairless Mice.

Dietary sphingolipids such as glucosylceramide and sphingomyelin are known to improve the skin barrier function of damaged skin. In this study, we focused on free-ceramide prepared from soy sauce lees, which is a byproduct of soy sauce production. The effects of dietary soy sauce lees ceramide on the skin of normal mice were evaluated and compared with those of dietary maize glucosylceramide. We found that transepidermal water loss value was significantly suppressed by dietary supplementation with soy sauce lees ceramide as effectively as or more effectively than maize glucosylceramide. Although the content of total and each subclass of ceramide in the epidermis was not significantly altered by dietary sphingolipids, that of 12 types of ceramide molecules, which were not present in dietary sources, was significantly increased upon ingestion of maize glucosylceramide and showed a tendency to increase with soy sauce lees ceramide intake. In addition, the mRNA expression of ceramide synthase 4 and involucrin in the skin was downregulated by sphingolipids. This study, for the first time, demonstrated that dietary soy sauce lees ceramide enhances skin barrier function in normal hairless mice, although further studies are needed to clarify the molecular mechanism.

Appearance of Intact Molecules of Dietary Ceramides Prepared from Soy Sauce Lees and Rice Glucosylceramides in Mouse Plasma.

Although the beneficial effects of dietary sphingolipids have recently been reported, the mechanism of their intestinal absorption has yet to be fully elucidated. In this study, the absorption and metabolism of dietary ceramides and glucosylceramides were evaluated by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis in the plasma of mice after a single oral administration. Ceramide molecules prepared from soy sauce lees (mainly composed of phytosphingosine and its derivatives) were undetectable or minor compounds in the plasma of control mice but appeared 1-6 h after administration. Rice glucosylceramide (mainly composed of sphingadienine) was endogenously detected in mouse plasma and showed a tendency to increase 1-6 h after administration by LC-MS/MS analysis. In addition, the ceramide molecules, which are hydrolysates of dietary glucosylceramide, were significantly increased in the plasma after administration. These findings strongly suggest that dietary ceramides and glucosylceramides are partly absorbed as intact molecules or hydrolysates.

Evaluation of Intestinal Absorption of Dietary Halocynthiaxanthin, a Carotenoid from the Sea Squirt Halocynthia roretzi.

Halocynthiaxanthin is an acetylenic carotenoid mainly found in Halocynthia roretzi. To date, several bioactivities of halocynthiaxanthin have been reported, but its mechanism of digestion and absorption in mammals has not been studied yet. In this study, we evaluated the intestinal absorption of halocynthiaxanthin in mice. The halocynthiaxanthin-rich fraction was prepared from the tunicate Halocynthia roretzi. Mice were orally administered the fraction at a dose of 5 mg/kg body weight. The halocynthiaxanthin levels in the plasma, liver, and small intestine, were quantified using HPLC-PDA, 1, 3, 6, and 9 h after ingestion. The halocynthiaxanthin-rich fraction mainly consisted of the all-trans form and a small amount of cis forms. These three isomers were detected in the plasma of mice 3 h after ingestion. Time-course changes after the ingestion of this fraction were found, with cis isomers being more abundant than the all-trans isomer in the mouse plasma and liver. In the small intestine, however, the all-trans isomer was primarily detected. The possibility that cis isomers might be absorbed rapidly from the small intestine cannot be denied, but our results suggest that dietary all-trans-halocynthiaxanthin might be isomerized to the cis isomer after intestinal absorption.

Dietary ceramide 2-aminoethylphosphonate, a marine sphingophosphonolipid, improves skin barrier function in hairless mice.

Sphingolipids are one of the major components of cell membranes and are ubiquitous in eukaryotic organisms. Ceramide 2-aminoethylphosphonate (CAEP) of marine origin is a unique and abundant sphingophosphonolipid with a C-P bond. Although molluscs such as squids and bivalves, containing CAEP, are consumed globally, the dietary efficacy of CAEP is not understood. We investigated the efficacy of marine sphingophosphonolipids by studying the effect of dietary CAEP on the improvement of the skin barrier function in hairless mice fed a diet that induces severely dry-skin condition. The disrupted skin barrier functions such as an increase in the transepidermal water loss (TEWL), a decrease in the skin hydration index, and epidermal hyperplasia were restored by CEAP dietary supplementation. Correspondingly, dietary CAEP significantly increased the content of covalently bound ω-hydroxyceramide, and the expression of its biosynthesis-related genes in the skin. These effects of dietary CAEP mimic those of dietary plant glucosylceramide. The novel observations from this study show an enhancement in the skin barrier function by dietary CAEP and the effects could be contributed by the upregulation of covalently bound ω-hydroxyceramide synthesis in the skin.

Absorption and Tissue Distribution of Siphonaxanthin from Green Algae.

Siphonaxanthin has been known to possess inhibitory effects against obesity, inflammation, and angiogenesis. However, little information on its in vivo bioavailability and biotransformation is available. To assess the bioavailability and metabolism of siphonaxanthin, its absorption and accumulation were evaluated using intestinal Caco-2 cells and Institute of Cancer Research (ICR) mice. Siphonaxanthin was absorbed and exhibited non-uniform accumulation and distribution patterns in tissues of ICR mice. Notably, in addition to siphonaxanthin, three main compounds were detected following dietary administration of siphonaxanthin. Because the compounds showed changes on mass spectra compared with that of siphonaxanthin, they were presumed to be metabolites of siphonaxanthin in ICR mice. Siphonaxanthin mainly accumulated in stomach and small intestine, while putative metabolites of siphonaxanthin mainly accumulated in liver and adipose tissues. Furthermore, siphonaxanthin and its putative metabolites selectively accumulated in white adipose tissue (WAT), especially mesenteric WAT. These results provide useful evidence regarding the in vivo bioactivity of siphonaxanthin. In particular, the results regarding the specific accumulation of siphonaxanthin and its metabolites in WAT have important implications for understanding their anti-obesity effects and regulatory roles in lipid metabolism.

Siphonaxanthin, a carotenoid from green algae Codium cylindricum, protects Ob/Ob mice fed on a high-fat diet against lipotoxicity by ameliorating somatic stresses and restoring anti-oxidative capacity.

Oxidative stress is implicated in the pathogenesis of many diseases including obesity, non-alcoholic fatty liver disease, and diabetes mellitus. Previously, we reported that siphonaxanthin, a carotenoid from green algae, elicited a potent inhibitory effect on hepatic de novo lipogenesis, and an anti-obesity effect in both 3T3L1 cells and KKAy mice. Thus, we hypothesized that consumption of siphonaxanthin could improve metabolic disorders including hepatic steatosis and systemic adiposity, as well as ameliorate somatic stress under obese conditions. Both the hepatocyte cell line HepG2 and a mouse model of severe obesity, produced by feeding Ob/Ob mice on a high-fat diet (HFD), were used to test this hypothesis. In obese mice, siphonaxanthin intake did not improve liver steatosis or systemic adiposity. However, intake did lower plasma glucose and alanine aminotransferase (ALT) levels and diminished hepatic lipid peroxidation products and antioxidant gene expression, which increased significantly in control group obese mice. Renal protein carbonyl content decreased significantly in the siphonaxanthin group, which might also indicate an ameliorated oxidative stress. Siphonaxanthin restored gene expression related to antioxidant signaling, lipid ß-oxidation, and endoplasmic-reticulum-associated protein degradation in the kidney, which decreased significantly in obese mice. Liver and kidney responded to obesity-induced somatic stress in a divergent pattern. In addition, we confirmed that siphonaxanthin potently induced Nrf2-regulated antioxidant signaling in HepG2 cells. In conclusion, our results indicated that siphonaxanthin might protect obesity-leading somatic stress through restoration of Nrf2-regulated antioxidant signaling, and might be a promising nutritional supplement.

Exopolysaccharides from milk fermented by lactic acid bacteria enhance dietary carotenoid bioavailability in humans in a randomized crossover trial and in rats.

BACKGROUND: Dietary supplementation with carotenoids can have beneficial health effects, but carotenoids are poorly absorbed. OBJECTIVES: We aimed to evaluate how milk fermented by lactic acid bacteria affects dietary carotenoid bioavailability in humans and rats and to investigate mechanisms by which active components in milk fermented by Lactobacilli enhance dietary carotenoid absorption. METHODS: Male rats (n = 8/group) were administered ß-carotene or ß-carotene + fermented milk. Rats (n = 6/group) were also pretreated with ezetimibe, a cholesterol absorption inhibitor, to investigate ß-carotene transport mechanisms. In humans, 3 studies were conducted using a randomized crossover method. Subjects (n = 16/study) consumed a vegetable (carrot, tomato, or spinach) drink alone or with a fermented milk drink. Blood samples were collected at various time points after consumption. RESULTS: In rats, the serum ß-carotene area under the concentration-time curve (AUC) was significantly higher for the ß-carotene + fermented milk than for ß-carotene only. A significant correlation (r = 0.83, P < 0.001) between the exopolysaccharide (EPS) content of fermented milk and serum ß-carotene AUC was observed. Ezetimibe treatment did not suppress elevations in serum ß-carotene concentrations induced by fermented milk ingestion. In humans, the incremental area under the concentration-time curve (iAUC) for ß-carotene in the plasma triacylglycerol-rich lipoprotein (TRL) fraction was significantly (1.8-fold, range: 0.6-3.9) higher when carrot + fermented milk was consumed compared with carrot drink alone. A significantly (6.5-fold, range: 0.04-7.7) higher iAUC for lycopene in the plasma TRL fraction was observed for subjects who consumed tomato + fermented milk compared with tomato drink alone. A significant increase in plasma lutein in all fractions was observed after consumption of spinach + fermented milk, but not with spinach drink alone. CONCLUSIONS: Co-ingestion of ß-carotene and fermented milk significantly increased dietary ß-carotene bioavailability in humans and rats. EPSs could affect the physical properties of fermented milk to enhance dietary ß-carotene absorption mediated by simple diffusion mechanisms. These findings may be relevant for methods to increase dietary carotenoid bioavailability.This trial was registered at umin.ac.jp/ctr as UMIN000034838, UMIN000034839, and UMIN000034840.

Gut microbial fatty acid metabolites (KetoA and KetoC) affect the progression of nonalcoholic steatohepatitis and reverse cholesterol transport metabolism in mouse model.

Nonalcoholic steatohepatitis (NASH) is a common liver disease that occurs in both alcoholics and nonalcoholics. Oxidative stress is a possible causative factor for liver diseases including NASH. Gut microorganisms, especially lactic acid bacteria, can produce unique fatty acids, including hydroxy, oxo, conjugated, and partially saturated fatty acids. The oxo fatty acid 10-oxo-11(E)-octadecenoic acid (KetoC) provides potent cytoprotective effects against oxidative stress through activation of Nrf2-ARE pathway. The aim of this study was to explore the preventive and therapeutic effects of gut microbial fatty acid metabolites in a NASH mouse model. The mice were divided into 3 experimental groups and fed as follows: (1) high-fat diet (HFD) (2) HFD mixed with 0.1% KetoA (10-oxo-12(Z)-octadecenoic acid), and (3) HFD mixed with 0.1% KetoC. After 3 weeks of feeding, plasma parameters, liver histology, and mRNA expression of multiple genes were assessed. There was hardly any difference in fat accumulation in the histological study; however, no ballooning occurred in 2/5 mice of KetoC group. Bridging fibrosis was not observed in the KetoA group, although KetoA administration did not significantly suppress fibrosis score (p = 0.10). In addition, KetoC increased the expression level of HDL related genes and HDL cholesterol levels in the plasma. These results indicated that KetoA and KetoC may partly affect the progression of NASH in mice models.

Siphonaxanthin, a carotenoid from green algae, suppresses advanced glycation end product-induced inflammatory responses.

Advanced glycation end products (AGEs) induce inflammation and contribute to the pathogenesis of atherosclerosis. Although many studies have demonstrated the protective effects of carotenoids against atherosclerosis, the effects of carotenoids on AGE-induced inflammation have not been characterized. As such, we aimed to identify carotenoids that provided protection against AGE-elicited inflammation. AGE-stimulated RAW264 macrophages were first evaluated for NO generation. Among 17 carotenoids tested, only siphonaxanthin significantly suppressed it. Next, mRNA expression levels were measured in RAW264 macrophages and human umbilical vascular endothelial cells following siphonaxanthin and AGE treatment. Siphonaxanthin significantly suppressed AGE-induced mRNA expression of interleukin-6 and cellular adhesion molecules, which are known to be important for the pathogenesis of atherosclerosis. Siphonaxanthin also significantly suppressed endoplasmic reticulum (ER) stress marker genes. A reporter gene assay revealed that siphonaxanthin, as well as an ER stress inhibitor, significantly inhibited AGE-induced nuclear factor-κB (NF-κB) activation. Altogether, mitigation of ER stress and subsequent NF-κB activation is one of the molecular mechanisms by which siphonaxanthin suppressed AGE-elicited inflammation. Siphonaxanthin is a carotenoid commonly found in standard diets and is considered relatively safe for human consumption, and hence, dietary intake of siphonaxanthin or siphonaxanthin-containing green algae could be beneficial in lowering the risk of developing atherosclerosis.