Caffeic acid-grafted chitooligosaccharides downregulate MAPK and NF-kB in RAW264.7 cells.

Chitooligosaccharide (COS) is a derivative of chitosan, which is a natural macromolecular compound. COS has been shown effects in an inflammatory response. Recent reports show that COS derivatives have enhanced anti-inflammatory activity by inhibiting intracellular signals. Evaluation of the anti-inflammatory effect of caffeic acid conjugated COS chain (CA-COS) was performed in this study. The effects of CA-COS on the inflammatory response were demonstrated in lipopolysaccharide-stimulated RAW264.7 macrophages. The results showed that CA-COS inhibited nitric oxide (NO) production and downregulated the gene expression of nitric oxide synthase (iNOS), and cytokines such as tumor necrosis factor-alpha (TNF-α), IL-1ß, and IL-6 without cytotoxic effect. In addition, western blot analysis showed that CA-COS inhibits the protein expression of iNOS and nuclear factor kappa B (NF-kB), including p50 and p65, and mitogen-activated protein kinase (MAPK) signaling pathways. Collectively, these results provide clear evidence for the anti-inflammatory mechanism of CA-COS that show great potential as a novel agent for the prevention and therapy of inflammatory diseases.

Possibility of nanostructured lipid carriers encapsulating astaxanthin from Haematococcus pluvialis to alleviate skin injury in radiotherapy.

PURPOSE: The study aimed to protect patients' skin against ionizing irradiation during radiotherapy by using astaxanthin-encapsulated nanostructured lipid carriers (NLC-ATX). MATERIALS AND METHODS: NLC-ATX was prepared by a combined method of hot homogenization and sonication. Cytotoxicity of NLC-ATX was evaluated by MTT colorimetric assay. The in vitro radioprotection of NLC-ATX for human fibroblast (HF) cells was investigated based on the level of ROS (reactive oxygen species), DNA damage, and cell death caused by X-irradiation. In addition, the in vivo radioprotection was evaluated based on the appearance and histological structure of the irradiated skin. RESULTS: NLC-ATX was successfully prepared, with a mean particle size, zeta potential, and encapsulation efficiency of 114.4 nm, -34.1 mV, and 85.67%, respectively. Compared to the control, NLC-ATX, at an optimum ATX concentration under in vitro condition, reduced the amount of generated ROS and DNA damage of 81.6% and 41.6%, respectively, after X-radiation, resulting in a significant decrease in cell death by 62.69%. Under in vivo condition, after the 9th day of X-irradiation (equivalent to an accumulated dose of 14 Gy), the dorsal skin of five out of six NLC-ATX-untreated mice exhibited grade-1 skin damage, according to CTCAE v5.0, while treatment with NLC-ATX protected 6/6 mice from acute skin damage. Moreover, on the 28th day after the first X-irradiation, the histological images illustrated that NLC-ATX at an ATX concentration of 0.25 µg/mL exhibited good recovery of the skin, with barely any difference noted in the collagen fibers and sebaceous glands compared to normal skin. CONCLUSIONS: NLC-ATX shows potential for application in skin protection against adverse effects of ionizing rays during radiotherapy.

Ferulic acid grafted onto chitooligosaccharides attenuates LPS-stimulated in murine macrophages by modulating the NF-κB and MAPK pathways.

Although chitooligosaccharides (COS) improve the drawbacks of chitosan, their biological activities in medical applications have not been highly appreciated. The main approach is to synthesise the COS derivatives in order to improve the biological properties of the COS. In this study, ferulic acid (FA) grafted onto COS (FA-COS) were synthesised and their mechanism of anti-inflammatory activity was investigated in the murine macrophage cells. The synthesis conditions of FA-COS were optimised and confirmed that the FA was successfully conjugated onto COS with the grafting effect of 15-34%. FA-COS exhibited anti-inflammatory activities via suppressing of nitric oxide formation, reducing iNOS expression at transcription and translation levels, down-regulation of TNF-α, IL-6 and IL-1 ß genes; NF-κB and MAPKs signalling pathways. These results show anti-inflammatory molecular mechanism of FA-COS that exhibit enormous potential for prevention of inflammatory diseases.

GABA-enriched rice bran inhibits inflammation in LPS-stimulated macrophages via suppression of TLR4-MAPK/NF-κB signaling cascades.

Gamma-aminobutyric acid (GABA)-enriched products (GEP) exhibited a wide range of pharmaceutical properties. In this study, anti-inflammatory activity of GEP from Lactobacillus fermentum-fermented water solution of rice bran was evaluated on lipopolysaccharide-activated macrophage model. GABA content in L. fermentum-fermented rice bran solution was determined up to 1.27 g/L. GEP was shown to inhibit the expression levels of inducible nitric oxide synthase and cyclooxygenase-2 enzymes. Moreover, pretreatment of GEP attenuated the generation level of interleukin (IL)-6, IL-1ß, tumor necrosis factor α, and monocyte chemoattractant protein-1. Especially, the activation of signaling pathways due to nuclear factor-κB (NF-κB) and mitogen-activated protein kinases (MAPKs) was interrupted in GEP-exposed cells. Notably, molecular docking result showed a potential binding of GABA to Toll-like receptor 4 with a binding energy of -3.88 kcal/mol, suggesting the role of GABA in suppression of Toll-like receptor 4-MAPK/NF-κB signaling cascades. As the result, GEP from L. fermentum-fermented rice bran solution could be suggested as a promising food for suppression of inflammatory responses. PRACTICAL APPLICATIONS: GABA-enriched products have been evidenced to possess various pharmaceutical properties and health beneficial effects. In this study, GABA-enriched product from L. fermentum-fermented rice bran solution exhibited the inhibition on inflammatory response in macrophages. Hence, it could be used as a potential ingredient for the mitigation of inflammatory responses.

Improving silymarin oral bioavailability using silica-installed redox nanoparticle to suppress inflammatory bowel disease.

Chronic inflammatory diseases such as inflammatory bowel diseases (IBD), which are strongly related to the overproduction of reactive oxygen species (ROS), have become more threatening to health. Silymarin is an active compound with the effect of expressing anti-inflammatory activity; however, it exhibits poor bioavailability due to the rapid metabolism and secretion, low permeability across the intestinal epithelial cells, and poor water solubility. In this study, we developed silica-containing redox nanoparticles (siRNP) with 50-60 nm in diameter to improve the bioavailability of silymarin by improving its uptake into the bloodstream and delivery to the targeted tissues of the colon. Silymarin-loaded siRNP (SM@siRNP) significantly increased the antioxidant capacity and anti-inflammatory efficacy in vitro by scavenging 2,2-diphenyl-1-picrylhydrazyl free radical and suppressing nitric oxide and pro-inflammatory cytokines as compared to the other treatments such as free silymarin, siRNP, and silymarin-loaded si-nRNP (the control nanoparticle without ROS scavenging property). Orally administered SM@siRNP significantly improved the bioavailability of silymarin and its retention in the colonic mucosa. The anti-inflammatory effects of SM@siRNP were also investigated in dextran sodium sulfate (DSS)-induced colitis in mice and it was observed that SM@siRNP treatment significantly improved the damage in the colonic mucosa of DSS colitis mice as compared to the other treatments. The results in this study indicate that SM@siRNP is a promising nanomedicine for enhancing the anti-inflammatory activity of silymarin and has a high potential for the treatment of IBD.

Draft genome sequence data of Rhodosporidium toruloides VN1, a strain capable of producing natural astaxanthin.

Rhodosporidium toruloides strain VN1 is of special interest because of its capability for high astaxanthin production. Here, we report the draft genome sequence of R. toruloides VN1, which comprises 20.01 Mb in 424 contigs with an overall G + C content of 61.8%. This whole-genome shotgun project has been deposited at DDBJ/EMBL/GenBank under the accession number SJTE00000000.

Poly(ornithine)-based self-assembling drug for recovery of hyperammonemia and damage in acute liver injury.

Oligo-peptides, including monomeric amino acids, have received much attention as bioactive molecules and drugs. One of the biggest problems of these compounds, however, is their very short bioavailability due to instant metabolism and rapid excretion. To solve this problem, we newly designed a poly(ethylene glycol) (PEG)-block-polypeptide self-assembling based drug for the treatment of acute liver injury. Here, PEG-block-poly(L-Ornithine) (PEG-b-POrn) was synthesized via a ring opening polymerization, and a nano-sized polyion self-assembling complex (NanoOrn) was prepared by simply mixing polycationic PEG-b-POrn with polyanionic chondroitin sulfate. The obtained NanoOrn was quite stable under high ionic strength and different pH conditions and NanoOrn exhibited extremely low toxicity in vitro and in vivo as compared to the original PEG-b-POrn. As compared to monomeric L-ornithine, administration of NanoOrn to mice significantly improved bioavailability of liberated ornithine, especially in the liver. Interestingly, NanoOrn treatment in acetaminophen (APAP)-induced acute liver injury mice remarkably suppressed blood ammonia levels and liver injury markers, resulting in more effective improvement of liver damage compared to monomeric ornithine via activation of ornithine transcarbomylase. These results show that the self-assembling polypeptide NanoOrn may provide a new concept and promising therapeutics as nanomedicines.

The Role of Rhodomyrtus tomentosa (Aiton) Hassk. Fruits in Downregulation of Mast Cells-Mediated Allergic Responses.

Rhodomyrtus tomentosa, a flowering plant of Myrtaceae family from southern and southeastern Asia, was known to possess a rich source of structurally diverse and various biological activities. In this study, the inhibitory effect of R. tomentosa fruit extract (RFE) on allergic responses in calcium ionophore A23187-activated RBL-2H3 mast cells was investigated. The result showed that RFE was able to inhibit mast cell degranulation via decreasing ß-hexosaminidase release and intracellular Ca2+ elevation at the concentration of 400 µg/ml. Moreover, the suppressive effects of RFE on the production of interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α) were evidenced. In addition, RFE effectively scavenged DPPH radical and suppressed the reactive oxygen species generation in a dose-dependent manner. Notably, the pretreatment of RFE caused the downregulation of tyrosine kinase Fyn phospholipid enzyme phospholipase Cγ (PLCγ), extracellular-signal-regulated kinase (ERK), and nuclear factor kappa B (NF-κB) phosphorylation. These results indicated that RFE could be a promising inhibitor of allergic responses and may be developed as bioactive ingredient for prevention or treatment of allergic diseases.

Investigation of Biological Activities of Wild Bitter Melon (Momordica charantia Linn. Var. Abbreviata Ser.).

Wild bitter melon (Momordica charantia L. var. Abbreviata Ser.) is a wild edible variety of M. charantia, often used in folk medicine. In this study, the biological activities of its extract and fractions were investigated in vitro. It was found that ethyl acetate (EA) fraction exhibited high 1,1-diphenyl-2-picryl-hydrazyl (DPPH) scavenging activity with a half maximal inhibitory concentration (IC50) value of 0.43 ± 0.04 mg/mL, while the chloroform (CF), EA, and n-butanol (Bu) fractions had strong 2,2-azinobis-3-ethyl benzothiazoline-6-sulfonic acid (ABTS)+ scavenging ability with IC50 values of 0.36 ± 0.04 mg/mL, 0.35 ± 0.02 mg/mL, and 0.35 ± 0.05 mg/mL, respectively. Moreover, the EA and Bu fractions exhibited the highest protective effect against H2O2-induced DNA damage in a concentration-dependent manner. Furthermore, the EA fraction was effective in the inhibition of enzyme α-amylase activity with an IC50 value of 0.27 ± 0.029 mg/mL. Finally, it was observed that the production of nitric oxide (NO), a pro-inflammatory mediator, was significantly reduced from LPS-stimulated murine macrophage RAW 264.7 cells by the ethanol extract (ET) and the EA fraction. Therefore, wild bitter melon could be considered as a promising biomaterial for the development of pharmaceutical products.

Antiproliferative Effect of Aminoethyl-Chitooligosaccharide on Human Lung A549 Cancer Cells.

The aminoethyl-chitooligosaccharide (AE-COS) was reported to inhibit human gastric cancer cell proliferation and human fibrosarcoma cell invasion. In this study, the role of AE-COS in down-regulation of proliferation of human lung A549 cancer cells was evaluated. It was found that AE-COS was able to reduce A549 cell proliferation to (32 ± 1.3)% at a concentration of 500 µg/ml. Moreover, AE-COS treatment caused suppression on COX-2 expression in a dose-dependent manner. Notably, the role of AE-COS in induction of cell apoptosis was observed via decreasing Bcl-2 expression and increasing caspase-3 and -9 activation. Accordingly, the antiproliferative effect of AE-COS was indicated due to suppression of cell proliferation and induction of cell apoptosis, suggesting AE-COS as a promising chemotherapy agent for treatment of lung cancer.

The Suppressive Activity of Fucofuroeckol-A Derived from Brown Algal Ecklonia stolonifera Okamura on UVB-Induced Mast Cell Degranulation.

UV light, especially UVB, is known as a trigger of allergic reaction, leading to mast cell degranulation and histamine release. In this study, phlorotannin Fucofuroeckol-A (F-A) derived from brown algal Ecklonia stolonifera Okamura was evaluated for its protective capability against UVB-induced allergic reaction in RBL-2H3 mast cells. It was revealed that F-A significantly suppress mast cell degranulation via decreasing histamine release as well as intracellular Ca2+ elevation at the concentration of 50 µM. Moreover, the inhibitory effect of F-A on IL-1ß and TNF-α productions was also evidenced. Notably, the protective activity of F-A against mast cell degranulation was found due to scavenging ROS production. Accordingly, F-A from brown algal E. stolonifera was suggested to be promising candidate for its protective capability against UVB-induced allergic reaction.

Inhibitory effect of aminoethyl-chitooligosaccharides on invasion of human fibrosarcoma cells.

Chitooligosaccharides (COS) have been reported to show a variety of biological efficacies such as anti-bacterial activity, anti-tumor activity and immune activity. The purpose of this study is to investigate the inhibitory effect of aminoethyl-chitooligosaccharides (AE-COS) synthesized from COS that were substituted hydroxyl groups with aminoethyl group at C-6 position on cell invasion of human fibrosarcoma cells. First of all, the effect of AE-COS on cell viability was observed using MTT assay. The cytotoxicity of AE-COS was increased in a dose dependent manner. The inhibitory effects of AE-COS on the activity and expression level of MMP-2 and MMP-9 related to invasion of cancer cells were examined using gelatin zymography and western blot. It was found that AE-COS above 20µg/ml showed the inhibitory effect on the activity and expression of MMP-9. Furthermore, AE-COS at 20µg/ml reduced the expression level of p50, a part of NF-κB, compared with phorbol-12- myristate-13- acetate (PMA) group. The available data let us hypothesize that AE-COS could provide chemoprevention as an inhibitor against cell invasion associated with metastasis.

Prevention of H2O2-induced oxidative stress in Chang liver cells by 4-hydroxybenzyl-chitooligomers.

In this study, a bioactive derivative of chitooligomers (1.0-3.0 kDa), 4-hydroxybenzyl-COS (HB-COS), was synthesized to enhance antioxidant activity. Hence, HB-COS was evaluated for its capabilities against H2O2-induced oxidative stress in human Chang liver cells. It was found that HB-COS possessed the free radical scavenging activity via decreasing the intracellular reactive oxygen species production. Furthermore, HB-COS significantly reduced the oxidation of DNA in a dose-dependent manner. Notably, HB-COS treatment upregulated the gene and protein expressions of antioxidative enzymes and thereby enhancing the intracellular antioxidant mechanisms. In addition, HB-COS treatment caused a remarkable blockade on degradation of inhibitory kappa B alpha (IκB-α) protein and translocation of nuclear factor kappa B (NF-κB). The current study demonstrated that HB-COS effectively attenuated hydrogen peroxide-induced oxidative stress in Chang liver cells by increasing levels of antioxidant enzymes and inhibiting reactive oxygen species generation, DNA oxidation and the NF-κB signaling pathway.

Protection of 4-hydroxybenzyl-chitooligomers against inflammatory responses in Chang liver cells.

The aim of this study was to investigate anti-inflammatory activity of 4-hydroxybenzyl-chitooligomers (HB-COS) in Chang liver cells stimulated by a cytokine mixture. It was revealed that HB-COS decreased the level of nitric oxide and prostaglandin E2 (PGE2) production by diminishing the expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) without significant cytotoxicity. Moreover, HB-COS exerted inhibitory effects on the production of pro-inflammatory mediator (interleukin-6) in Chang liver cells. Notably, HB-COS exhibited anti-inflammatory activities via blocking degradation of inhibitory kappa B alpha (IκB-α), translocation of nuclear factor kappa B (NF-κB), and phosphorylation of mitogen-activated protein kinases (MAPKs) in a dose-dependent manner. Collectively, these findings indicated that HB-COS possessed potential anti-inflammatory effects in Chang liver cells, and could be a useful therapeutic agent for the treatment of hepatic inflammatory diseases.

Biological activities and potential health benefits of bioactive peptides derived from marine organisms.

Marine organisms have been recognized as rich sources of bioactive compounds with valuable nutraceutical and pharmaceutical potentials. Recently, marine bioactive peptides have gained much attention because of their numerous health beneficial effects. Notably, these peptides exhibit various biological activities such as antioxidant, anti-hypertensive, anti-human immunodeficiency virus, anti-proliferative, anticoagulant, calcium-binding, anti-obesity and anti-diabetic activities. This review mainly presents biological activities of peptides from marine organisms and emphasizing their potential applications in foods as well as pharmaceutical areas.

Protective effects of aminoethyl-chitooligosaccharides against oxidative stress in mouse macrophage RAW 264.7 cells.

The aim of this study is to investigate the inhibitory effects of aminoethyl-chitooligosaccharides (AE-COS) on oxidative stress in mouse macrophages (RAW 264.7 cells). The inhibitory effects of AE-COS on DNA and protein oxidation were studied in RAW 264.7 cells. Furthermore, free radical scavenging effect of AE-COS were determined in RAW264.7 cells by 2',7'-dichlorofluorescein (DCF) intensity and intracellular glutathione (GSH) level. AE-COS also inhibited myeloperoxidase (MPO) activity in human myeloid cells (HL-60). These results suggest that AE-COS acts as a potential free radical scavenger in RAW 264.7 cells.