Sang Hyun Moh

IRAK1/4-Targeted Anti-Inflammatory Action of Caffeic Acid

Caffeic acid (CA) is a phenolic compound that is frequently present in fruits, grains, and dietary supplements. Although CA has been reported to display various biological activities such as anti-inflammatory, anti-cancer, anti-viral, and anti-oxidative effects, the action mechanism of CA is not yet fully elucidated. In this study, the anti-inflammatory action mechanism of CA was examined in lipopolysaccharide (LPS) treated macrophages (RAW264.7 cells) and HCl/EtOH-induced gastritis. CA was found to diminish nitric oxide (NO) and prostaglandin E2 (PGE2) production in LPS-stimulated RAW264.7 cells. Additionally, mRNA levels of tumor necrosis factor (TNF)-α, cyclooxygenase (COX)-2, and inducible NO synthase (iNOS) were downregulated by CA. CA also strongly suppressed the nuclear translocation of AP-1 family proteins and the related upstream signaling cascade composed of interleukin-1 receptor-associated kinase 1 (IRAK1), IRAK4, TGF-β-activated kinase 1 (TAK1), mitogen-activated protein kinase kinase 4/7 (MKK4/7), and c-Jun N-terminal kinase (JNK). In a direct kinase assay, CA was revealed to directly inhibit IRAK1 and IRAK4. CA also ameliorated HCl/EtOH-induced gastric symptoms via the suppression of JNK, IRAK1, and IRAK4. Therefore, our data strongly suggest that CA acts as an anti-inflammatory drug by directly suppressing IRAK1 and IRAK4.

Methanol extract of Osbeckia stellata suppresses lipopolysaccharide- and HCl/ethanol-induced inflammatory responses by inhibiting Src/Syk and IRAK1

ETHNOPHARMACOLOGICAL RELEVANCE Osbeckia stellata Buch.-Ham. ex D.Don is traditionally prescribed to treat various inflammatory diseases. However, how this plant is able to modulate inflammatory responses is unknown. This study explored the anti-inflammatory effects of 99% methanol extracts of O. stellata (Os-ME). MATERIALS AND METHODS The anti-inflammatory effect of Os-ME was evaluated by measuring the levels of nitric oxide (NO) and prostaglandin E(2) (PGE(2)) in lipopolysaccharide (LPS)-treated RAW264.7 macrophage cells and by determining gastric inflammatory lesions in mice induced by HCl/ethanol (EtOH). The molecular mechanisms of the inhibitions were elucidated by analyzing the activation of transcription factors, upstream signaling cascade, and the kinase activities of target enzymes. RESULTS Os-ME dose-dependently diminished the release of NO and PGE(2), and suppressed the expression of inducible NO synthase and cyclooxygenase-2 in LPS-treated RAW264.7 cells. Os-ME clearly inhibited the translocation of c-Rel, a subunit of nuclear factor κB (NF-κB), and c-Fos, a subunit of activator protein-1 (AP-1), and their regulatory upstream enzymes including Src, Syk, and IRAK1. Interestingly, orally administered Os-ME ameliorated acute inflammatory symptoms and suppressed the activation of Src, Syk, and IRAK1 induced by HCl/EtOH treatment in mouse stomach. CONCLUSION Os-ME can be considered as an orally available anti-inflammatory herbal remedy with Src/Syk/NF-κB and IRAK1/AP-1 inhibitory properties.

Src and Syk are targeted to an anti-inflammatory ethanol extract of Aralia continentalis

ETHNOPHARMACOLOGICAL RELEVANCE Aralia continentalis Kitagawa (Araliaceae) is a representative ethnomedicinal herbal plant traditionally prescribed in Korea to relieve various inflammatory symptoms. However, the exact molecular mechanism of its anti-inflammatory activity has not been fully investigated. MATERIALS AND METHODS The effect of the ethanol extract from the roots of this plant (Ac-EE) on the production of the inflammatory mediator nitric oxide (NO) was studied in RAW264.7 cells. Its effect on inflammatory symptoms (gastritis and hepatitis) in mice was also examined. In particular, the molecular inhibitory mechanism was analysed by measuring the activation of transcription factors and their upstream signalling and the kinase activity of target enzymes. RESULTS Ac-EE dose-dependently suppressed NO production in lipopolysaccharide (LPS)-activated RAW264.7 cells. This extract also displayed curative activity against EtOH/HCl-induced gastritis and LPS-induced hepatitis in mice. Ac-EE-mediated anti-inflammatory activity was found to be at the transcriptional level, as it blocked the activation of the nuclear factor (NF)-κB pathway composed of Syk and Src, according to immunoblotting and immunoprecipitation analyses and a kinase assay with whole and nucleus lysates from RAW264.7 cells and mice. CONCLUSION Ac-EE may be developed as a functional herbal remedy targeting Syk- and Src-mediated anti-inflammatory mechanisms. Future work using pre-clinical studies will be needed to investigate this possibility.

HangAmDan-B, an Ethnomedicinal Herbal Mixture, Suppresses Inflammatory Responses by Inhibiting Syk/NF-κB and JNK/ATF-2 Pathways

HangAmDan-B (HAD-B) is a powdered mixture of eight ethnopharmacologically characterized folk medicines that is prescribed for solid masses and cancers in Korea. In view of the finding that macrophage-mediated inflammation is a pathophysiologically important phenomenon, we investigated whether HAD-B modulates inflammatory responses and explored the associated molecular mechanisms. The immunomodulatory activity of HAD-B in toll-like receptor-activated macrophages induced by lipopolysaccharide (LPS) was assessed by measuring nitric oxide (NO) and prostaglandin E(2) (PGE(2)) levels. To identify the specific transcription factors (such as nuclear factor [NF]-κB and signaling enzymes) targeted by HAD-B, biochemical approaches, including kinase assays and immunoblot analysis, were additionally employed. HAD-B suppressed the production of PGE(2) and NO in LPS-activated macrophages in a dose-dependent manner. Furthermore, the extract ameliorated HCl/EtOH-induced gastritis symptoms. Moreover, HAD-B significantly inhibited LPS-induced mRNA expression of inducible NO synthase and cyclooxygenase (COX)-2. Interestingly, marked inhibition of NF-κB and activating transcription factor was observed in the presence of HAD-B. Data from direct kinase assays and immunoblot analysis showed that HAD-B suppresses activation of the upstream signaling cascade involving spleen tyrosine kinase, Src, p38, c-Jun N-terminal kinase, and transforming growth factor β-activated kinase 1. Finally, kaempferol, but not quercetin or resveratrol was identified as a bioactive compound in HAD-B. Therefore, our results suggest that HAD-B possesses anti-inflammatory activity that contributes to its anticancer property.

Development of Anti-Wrinkle Materials using Galloyl-Peptide Derivatives

Conjugating a phytochemical, a strong antioxidant, with a functional peptide not only compensates for its stability, but also improves its solubility and anti-wrinkle effects, thereby contributing to the possibility of becoming an excellent cosmetic ingredient. Thus, in this study we examined the potential cosmetic use of a phytochemical-peptide derivative using gallic acid, a phytochemical with antioxidant, anti-inflammatory, and anti-cancer effects. To evaluate the antioxidant and wrinkle-improving efficacy of 5 synthesized gallic acids conjugated with LVH, IVH, KTTKS, YGGFM, and YGGFLRKYP respectively, we observed the expression of genes related to wrinkle improvement using DPPH radical scavenging activity and real-time PCR. As a result, all 5 derivatives had excellent free radical scavenging effects. The expression level of genes involved collagen synthesis also increased, and the secreted peptides during collagen production contributed to their antioxidant and wrinkle improving effects. These results mark the potential use of gallic acid peptide derivatives as a cosmetic ingredient for anti-oxidation and wrinkle improvement.

Phenolic Contents and Antioxida nt Activities of Six Edible Seaweeds

Abstract Phenolic contents and antioxidant activities were determined in the water and ethanol extracts from six species ofedible seaweeds, Phaeophyceae ( Laminaria japonica, Hizikia fusiformis and Undaria pinnatifida ), Rhodophyceae ( Porphyra tenera and Gracilaria verrucosa ) and Chlorophyceae ( Ulva lactuca ). The highest extraction yield was observed in water extract (pH 8.0) of G. verrucosa (44.23 %) and phenolic content was the highest in ethanolic (75%) extract of H. fusiformis (52.82 μg/mg). 25 % ethanolic extract from U. lactuca was found to have the highest DPPH radicals scavenging activity(19.29 %) and superoxide anion scavenging activitiy was higher in water extract (pH 8.0) from H. fusiformis (81.20 %) than that other seaweeds. Phenolic contents were strongly correlated with antioxidant activity in the six edible seaweeds extracts (R 2 =0.852). Taken together, these results indicate that H fusiformis may be a excellentsource for development of natural antioxidants. Key Words :

Antiaging Effects of Algae-Derived Mycosporine-Like Amino Acids (MAAs) on Skin

Skin aging is a complex biological process that is a consequence of both intrinsic and extrinsic aging. Intrinsic aging refers to genetically programmed aging with time. Extrinsic aging is mainly caused by environmental factors such as ultraviolet damage, pollution, harsh weather, and cigarette smoke. Chronic sun exposure is one of the main causes of extrinsic skin aging and is responsible for age-related changes such as wrinkles, roughness, mottled hyperpigmentation, dilated blood vessels, and loss of skin tone. The substantial loss in the stratospheric ozone layer and consequent increment in solar ultraviolet (UV) radiation on the earth’s surface have augmented the interest in searching for natural photoprotective compounds. Several photosynthetic marine organisms have evolved defense mechanisms to counteract UV radiation by synthesizing UV-absorbing compounds, such as mycosporine-like amino acids (MAAs). MAAs have been reported in diverse organisms; they are a family of secondary metabolites that directly or indirectly absorb the energy of solar radiation and protect organisms from enhanced solar UV radiation. MAAs havemaximumUVabsorption between 310 and 362 nm and high molar extinction coefficients. In addition, they have a capability to dissipate absorbed radiation as heat efficiently without producing reactive oxygen species (ROS). And also MAAs increase the photostability and resistance to several abiotic stressors. Several MAAs are introduced in this chapter and their functional roles were suggested for antiaging. Among them, we will discuss three specific MAAs, porphyra-334, shinorine, and mycosporine-glycine (M-Gly), on their antiaging effects comprising antioxidant, antiinflammation, and skin-firming properties.

Effects of Hypoxia on the Fertilization and Early Development of Sea Urchin, Strongylocentrotus nudus

Dissolved oxygen is one of the most important factors controlling growth in aquatic organisms. Hypoxia is generally defined as dissolved oxygen less than 2.8 mg (equivalent to 2 mL or 91.4 mM). Therefore, hypoxia zone can cause a serious problem in marine ecosystem. In this study, to investigate embryotoxic (fertilization and embryo development rates) effects of hypoxia on sea urchin Strongylocentrotus nudus were exposed to dissolved oxygen levels of 7.6 mg (normoxia) and 1.8 mg (hypoxia) for 2 days at and 33 ‰. Also, Expression levels of stress related gene (HSP70) and antioxidant related gene (glutathione reductase) in the sea urchins exposed to hypoxia were confirmed by Immunoblotting and RT-PCR analysis. In results, we showed that developmental rates were dramatically reduced in hypoxia condition. Molecular analysis demonstrated that higher HSP70 (5.5 fold) and glutathione reductase gene (2.79 fold) were present in the sea urchin exposed to hypoxia. Our results suggested that hypoxia can cause the abnormal development and elicits a stress and antioxidant response on sea urchin.

Development of DNA chip for verification of 25 microalgae collected from southern coastal region in Korea

Countless species occur in the marine microalgal domain. Some are used as health functional foods or medical products but many species are harmful such as those that cause the red tide. Therefore, it is necessary to conduct prompt and accurate identification of microalgal species. As it is quite difficult to accurately distinguish all species in terms of morphology, we performed DNA barcoding analysis using molecular markers for more accurate and rapid screening. DNA barcoding analysis, i.e., DNA chip technology, is a powerful method for studies on microalgal taxonomy and biodiversity. We used the mitochondrial cytochrome c oxidase subunit I (mtCOI) as a barcoding gene to identify microalgal species. In this study, the diversity and phylogenetic differences among different microalgae were analyzed. Additionally, a microalgal species-specific probe was screened by 21–23 bp and the result was printed on silylated slide for use in a robotic microarrayer. As a result, we performed a DNA chip assay for each of 25 microalgal species and determined that the COI barcode gene was suitable as a marker gene, as it could identify various microalgae from the Korean South Sea by species.

Production Yield Enhancement of Mycosporine-like amino acid(MAA)s in Transformed Microalgae Culture by Radiofrequency

In sea water, microalgae are exposed to a range of critical environmental conditions. Microalgae are protected from UV-A radiation due to the presence of mycosporine like amino acids(MAAs). Owing to the UV-A absorption properties of MAAs, they are used widely as a UV protecting ingredient in cosmetics. Therefore, there is a need to increase the production yield of MAAs. This study investigated the production yield of MAAs in transformed microalgae by radiofrequency(RF) exposure. Initially, the Glut-1 gene was transformed to Chlamydomonas hedleyi microalgae as a glucose transporter. The biomass was enhanced after Glut-1 gene transformation. In addition, the MAAs production yield was increased during large scale production in bioreactors due to the RF treatment. Therefore, purified extracts of MAAs can be used as a sun block material in the cosmetic industrial field.