WonWoo Lee

Gallic acid isolated from Spirogyra sp. improves cardiovascular disease through a vasorelaxant and antihypertensive effect

In this study, we investigated the vasorelaxant and antihypertensive effects of gallic acid (GA), a polyphenol isolated from the green alga Spirogyra sp., to assess its suitability as a therapeutic for cardiovascular diseases (CVDs). We examined the effect of GA on endothelium-dependent vasorelaxation in human umbilical vein endothelial cells (HUVECs). GA increased nitric oxide (NO) levels by increasing phosphorylation of endothelial nitric oxide synthase (eNOS), and its effect on NO production was attenuated by pretreatment with the eNOS inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME). We also investigated its antihypertensive effect by examining GA-mediated inhibition of angiotensin-I converting enzyme (ACE). GA inhibited ACE with a half-maximal inhibitory concentration (IC50) value of 37.38 ± 0.39 μg/ml. In silico simulations revealed that GA binds to the active site of ACE (PDB: 1O86) with a binding energy of -270.487 kcal/mol. Furthermore, GA clearly reduced blood pressure in spontaneously hypertensive rats (SHR) to an extent comparable to captopril. These results suggest that GA isolated from Spirogyra sp. exerts multiple therapeutic effects and has potential as a CVD treatment.

Anti-inflammatory activity of halogenated secondary metabolites of Laurencia snackeyi (Weber-van Bosse) Masuda in LPS-stimulated RAW 264.7 macrophages

Secondary metabolites of tropical seaweed are proven to exhibit variety of biological activities. Six species of seaweed (Caulerpa racemosa var. laete-virens, Caulerpa sertularioides f. longipes, Halymenia dilatata, Laurencia snackeyi, Padina boryana, and Sargassum swartzii) were tested for anti-inflammatory activity in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Crude L. snackeyi extract exhibit potent activity, and upon bioassay-guided isolation, it contained four halogenated compounds that exert profound inhibitory effects against nitric oxide (NO) production in LPS-stimulated RAW 264.7 cells. These compounds were subjected to spectroscopic measurements and were identified as palisadin A (1), aplysistatin (2), 5-acetoxypalisadin B (3), and palisol (4). Further experiments showed aplysistatin (2) to significantly inhibit NO production and prostaglandin-E2 (PGE2) production, and suppress inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression in LPS-stimulated RAW 264.7 cells. Therefore, aplysistatin (2) is suggested to inhibit NO and PGE2 production via the inhibition of iNOS and COX-2, indicating that its activity may be attributed to the modulation of anti-inflammatory agents.

Assessment of anti-inflammatory effect of 5β-hydroxypalisadin B isolated from red seaweed Laurencia snackeyi in zebrafish embryo in vivo model

5β-Hydroxypalisadin B, a halogenated secondary metabolite isolated from red seaweed Laurencia snackeyi was evaluated for its anti-inflammatory activity in lipopolysaccharide (LPS)-induced zebrafish embryo. Preliminary studies suggested the effective concentrations of the compound as 0.25, 0.5, 1 μg/mL for further in vivo experiments. 5β-Hydroxypalisadin B, exhibited profound protective effect in the zebrafish embryo as confirmed by survival rate, heart beat rate, and yolk sac edema size. The compound acts as an effective agent against reactive oxygen species (ROS) formation induced by LPS and tail cut. Moreover, 5β-hydroxypalisadin B effectively inhibited the LPS-induced nitric oxide (NO) production in zebrafish embryo. All the tested protective effects of 5β-hydroxypalisadin B were comparable to the well-known anti-inflammatory agent dexamethasone. According to the results obtained, 5β-hydroxypalisadin B isolated from red seaweed L. snackeyi could be considered as an effective anti-inflammatory agent which might be further developed as a functional ingredient.

Protective effect of marine brown algal polyphenols against oxidative stressed zebrafish with high glucose

The zebrafish (Danio rerio) is one of the most widely used vertebrate models in research studies in molecular genetics, development biology, drug discovery and human disease. This study has confirmed an increase in the production of reactive oxygen species (ROS) and induction of cell death by high glucose treatment in zebrafish. We observed that exposure to phlorotannins, which include 6,6-bieckol, phloroeckol, dieckol and phlorofucofuroeckol isolated from an edible brown alga, Ecklonia cava, significantly inhibited high glucose induced ROS and cell death. Among the phlorotannins, DK (Dieckol) significantly reduced heart rates, ROS, nitric oxide (NO), lipid peroxidation generation and cell death in high glucose induced oxidative stress. Further, high glucose levels induced the over expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), whereas DK treatment reduced its over expression. These findings indicate that the zebrafish model is an efficient animal model that can be used to investigate hyperglycemia-stimulated oxidative stress. Therefore, this model can be used as an in vivo experiment to confirm the antioxidant properties of functional foods and nutraceuticals.

Radio-protective effect of polysaccharides isolated from Lactobacillus brevis-fermented Ecklonia cava

We investigated the radioprotective effects of a polysaccharide isolated from enzymatic extracts of Ecklonia cava (E. cava) fermented by fungi and bacteria. We identified that the aqueous extract of the Lactobacillus brevis-fermented E. cava especially showed the highest proliferation effect. In addition, the enzymatic extract prepared by enzyme-assisted extraction using Viscozyme (VLFE) significantly increased cell proliferation. Further study indicated that the polysaccharides isolated from the >30 kDa fraction of VLFE (VLFEP) significantly enhanced survival and proliferation effects in γ-ray-irradiated cells. Also, VLFEP markedly reduced the DNA damage, production of reactive oxygen species, and the percentage of Sub-G(1) DNA contents caused by γ-ray-irradiation. Moreover, VLFEP modulated the expression levels of p53, Bax, and Bcl-2 via inhibition of IκBα degradation and phosphorylation and NFκB p65 translocation into nuclei. These results demonstrate that VLFEP has radioprotective properties including the modulation of apoptosis via the inhibition of the NFκB signaling pathway.

A fucoidan fraction purified from Chnoospora minima; a potential inhibitor of LPS-induced inflammatory responses

Fucoidans are an interesting group of bioactive sulfated polysaccharides abundant in brown seaweeds. The current study highlights the enrichment and extraction of fucoidan from Chnoospora minima by means of enzyme-assistant extraction using Celluclast and evaluation of its anti-inflammatory potential through in vitro and in vivo studies. The purified C. minima fucoidan (F2,4) inhibited the nitrous oxide (NO) production (IC50=27.82±0.88μg/ml) and expression of PGE2 through the subsequent downregulation of iNOS and COX-2 expression in lipopolysaccharide (LPS) stimulated RAW 264.7 macrophages. F2,4 downregulated TNF-α, IL1-β, and IL-6 in RAW 264.7 macrophages in a dose-dependent manner and suppressed NO and ROS production in LPS stimulated zebrafish embryos while exerting a protective effect against the cell damage caused by LPS. Polysaccharide structural characterization was performed using FTIR, HPAE-PAD analysis of the monosaccharide content and NMR spectroscopy. Current findings confirm the potential anti-inflammatory activity of fucoidan purified from C. minima and elaborate its potential application as a functional ingredient in consumer products.

Antibacterial effect of citrus press-cakes dried by high speed and far-infrared radiation drying methods

In this study, the antibacterial effect was evaluated to determine the benefits of high speed drying (HSD) and far-infrared radiation drying (FIR) compared to the freeze drying (FD) method. Citrus press-cakes (CPCs) are released as a by-product in the citrus processing industry. Previous studies have shown that the HSD and FIR drying methods are much more economical for drying time and mass drying than those of FD, even though FD is the most qualified drying method. The disk diffusion assay was conducted, and the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined with methanol extracts of the dried CPCs against 11 fish and five food-related pathogenic bacteria. The disk diffusion results indicated that the CPCs dried by HSD, FIR, and FD prevented growth of all tested bacteria almost identically. The MIC and MBC results showed a range from 0.5-8.0 mg/mL and 1.0-16.0 mg/mL respectively. Scanning electron microscopy indicated that the extracts changed the morphology of the bacteria cell wall, leading to destruction. These results suggest that CPCs dried by HSD and FIR showed strong antibacterial activity against pathogenic bacteria and are more useful drying methods than that of the classic FD method in CPCs utilization.

The roles of NF-κB and ROS in regulation of pro-inflammatory mediators of inflammation induction in LPS-stimulated zebrafish embryos

Abstract In this study, the roles of reactive oxygen species (ROS) and NF‐&kgr;B on inflammation induction in lipopolysaccharide (LPS)‐stimulated zebrafish embryos were evaluated using N‐acetyl‐l‐cysteine (NAC) and pyrrolidine dithiocarbamate (PDTC), specific inhibitors of ROS and NF‐&kgr;B, respectively. LPS‐stimulated zebrafish embryos showed increasing production of NO and ROS and expression of iNOS and COX‐2 protein, compared to a control group without LPS. However, NAC significantly inhibited production of NO and ROS and markedly suppressed expression of iNOS and COX‐2 protein in LPS‐stimulated zebrafish embryos. The mRNA expressions of NF‐&kgr;B such as p65NF‐&kgr;B and I&kgr;B‐A were significantly increased after LPS stimulation, whereas PDTC attenuated mRNA expression of NF‐&kgr;B. PDTC also inhibited production of NO and reduced expression of iNOS and COX‐2 protein in LPS‐stimulated zebrafish embryos. Taken together, these results indicated that LPS increases pro‐inflammatory mediators in zebrafish embryos through ROS and NF‐&kgr;B regulation. HighlightsWe confirmed the roles of ROS and NF‐&kgr;B in LPS‐stimulated zebrafish embryos.We employed NAC and PDTC, which are specific inhibitors of ROS and NF‐kB.NAC inhibited LPS‐induced NO and ROS production and iNOS and COX‐2 expression.PDTC also inhibited LPS‐induced NO production and iNOS and COX‐2 expression.PDTC significantly inhibited LPS‐induced increases in the mRNA expression of NF‐&kgr;B.

Apoptotic and antiproliferative properties of 3β‐hydroxy‐Δ5‐steroidal congeners from a partially purified column fraction of Dendronephthya gigantea against HL‐60 and MCF‐7 cancer cells

Organisms belonging to the genus Dendronephthya are among a group of marine invertebrates that produce a variety of terpenoids with biofunctional properties. Many of these terpenoids have been proven effective as anticancer drugs. Here, we report the antiproliferative effect of 3β‐hydroxy‐Δ5‐steroidal congeners against the proliferation of HL‐60 human leukemia cells and MCF‐7 human breast cancer cells. The sterol‐rich fraction (DGEHF2‐1) inhibited the growth of HL‐60 and MCF‐7 cells with IC50 values of 13.59 ± 1.40 and 29.41 ± 0.87 μg ml–1 respectively. Treatment with DGEHF2‐1 caused a dose‐dependent increase in apoptotic body formation, DNA damage and the sub‐G1 apoptotic cell population. Moreover, DGEHF2‐1 downregulated the expression of Bcl‐xL while upregulating Bax, caspase‐9, and PARP cleavage in both HL‐60 and MCF‐7 cells. The steroid fraction was found to act via the mitochondria‐mediated apoptosis pathway. Identification of the sterols was performed via gas chromatography–tandem mass spectrometry analysis. Studying the mechanism of the anticancer effect caused by these sterol derivatives could lead to the identification of other natural products with anticancer properties.

Anti-inflammatory effects of trans-1,3-diphenyl-2,3-epoxypropane-1-one in zebrafish embryos in vivo model.

In this study, trans-1,3-diphenyl-2,3-epoxypropane-1-one (DPEP) was evaluated for its anti-inflammatory activity in lipopolysaccharide (LPS)-stimulated zebrafish embryos. In the present study, DPEP exhibited potential protective effect in the zebrafish embryos as confirmed by survival rate. DPEP acts as an effective agent against reactive oxygen species (ROS) formation induced by LPS. Moreover, DPEP effectively inhibited LPS-induced nitric oxide (NO) production in zebrafish embryos. In addition, DPEP significantly reduced the expression of inducible NOS (iNOS) and cycloxygenase 2 (COX-2), which generate NO as a key mediator of inflammation in a concentration-dependent manner. According to these results, DPEP could be considered an effective anti-inflammatory agent, which might be further developed as a functional ingredient. This is the first report of the anti-inflammatory activity of DPEP in the LPS-stimulated zebrafish model.