Libanoridin Isolated from Corydalis heterocarpa Inhibits Adipogenic Differentiation of Bone Marrow-Derived Mesenchymal Stromal Cells.

Bone marrow adiposity is a complication in osteoporotic patients. It is a result of the imbalance between adipogenic and osteogenic differentiation of bone marrow cells. Phytochemicals can alleviate osteoporotic complications by hindering bone loss and decreasing bone marrow adiposity. Corydalis heterocarpa is a biennial halophyte with reported bioactivities, and it is a source of different coumarin derivatives. Libanoridin is a coumarin isolated from C. heterocarpa, and the effect of libanoridin on adipogenic differentiation of human bone marrow-derived mesenchymal stromal cells (hBM-MSCs) was evaluated in the present study. Cells were induced to undergo adipogenesis, and their intracellular lipid accumulation and expression of adipogenic markers were observed under libanoridin treatment. Results showed that 10 µM libanoridin-treated adipocytes accumulated 44.94% less lipid compared to untreated adipocytes. In addition, mRNA levels of PPARγ, C/EBPα, and SREBP1c were dose-dependently suppressed with libanoridin treatment, whereas only protein levels of PPARγ were decreased in the presence of libanoridin. Fluorescence staining of adipocytes also revealed that cells treated with 10 µM libanoridin expressed less PPARγ compared to untreated adipocytes. Protein levels of perilipin and leptin, markers of mature adipocytes, were also suppressed in adipocytes treated with 10 µM libanoridin. Analysis of MAPK phosphorylation levels showed that treatment with libanoridin inhibited the activation of p38 and JNK MAPKs observed by decreased levels of phosphorylated p38 and JNK protein. It was suggested that libanoridin inhibited adipogenic differentiation of hBM-MSCs via suppressing MAPK-mediated PPARγ signaling. Future studies revealing the anti-adipogenic effects of libanoridin in vivo and elucidating its action mechanism will pave the way for libanoridin to be utilized as a nutraceutical with anti-osteoporotic properties.

Effect of organic acids on the formation of biogenic amines in fermented anchovy sauce comprising raw anchovy materials with different levels of freshness.

We investigated the effect of different levels of organic acids on the formation of biogenic amines in anchovy fish sauce. Fish sauce samples were prepared with fresh anchovies used immediately after being caught (F), and anchovies left at ambient temperature for 4 h (4 h), 18 h (18 h), or 24 h (24 h). Anchovies from each of the four groups were mixed with salt at a 4:1 ratio and then fermented at ambient temperature for varying periods of time. The F and 4 h anchovies contained higher levels of acetic acid, succinic acid, and lactic acid, and the levels increased during the fermentation process. The histamine content of the fish sauce samples prepared using F anchovies ranged from 44.0 to 9.2 mg/kg at 1 and 24 months of fermentation. The histamine content of fish sauce samples prepared using 4 h, 18 h, and 24 h anchovies was 111.5-37.0 mg/kg (4 h), 780.1-560.3 mg/kg (18 h), and 880.6-675.7 mg/kg (24 h). Our findings indicated that the histamine and other biogenic amine levels were closely associated with the degree of freshness of the raw anchovy material. These findings indicate that lower pH levels resulting from organic acids generated by the fresh raw material can effectively inhibit histamine formation.

Anti-obesity effects of Laminaria japonica fermentation on 3T3-L1 adipocytes are mediated by the inhibition of C/EBP-α/ß and PPAR-γ.

Obesity is global problem that contributes to disease, and is partly caused by fast-food, high-fat diets. Much attention has been focused on developing anti-obesity foods and chemical materials from natural sources. Seaweed has bioactive properties that influence immune activity and have anti-cancer and anti-obesity effects. Laminaria japonica is a widely consumed seaweed, and has been promoted as a health food in Korea. The bioactive properties of L. japonica include anti-cancer, anti-diabetic, and anti-inflammation effects. Most Laminaria japonica are distributed in a simple processing form such as drying, and their availability is very low. Therefore, various types of functional products can be developed if they can be applied to foods through functionalization using fermentation techniques. It is a structural problem that is the most problematic in seaweed processing. In this study, we used fermented Laminaria japonica. To increase physiological activity, fermentation treatment was performed to loosen the structure, thereby increasing the activity of the glycoprotein. First, we screened the anti-obesity potential of an L. japonica fermentation extract (LJF) using 3T3-L1 adipocyte cells. We determined cytotoxicity using an MTS assay and measured LJF for its ability to affect adipogenesis through glucose uptake, triglyceride levels, and Oil Red O staining. We confirmed that LJF inhibited adipocyte differentiation. CCAAT/enhancer-binding proteins α/ß (C/EBP-α/ß) and peroxisome proliferator-activated receptor-γ (PPAR-γ) are involved in the early and late stages of adipocyte differentiation. LJF significantly reduced the expression levels of C/EBP-α/ß and PPAR-γ and decreased the concentration of adiponectin. Thus, our results suggest that LJF inhibits adipogenesis in 3T3-L1 cells, and may be valuable for its anti-obesity effects.

Dioxinodehydroeckol Enhances the Differentiation of Osteoblasts by Regulating the Expression of Phospho-Smad1/5/8.

Lack of bone formation-related health problems are a major problem for the aging population in the modern world. As a part of the ongoing trend of developing natural substances that attenuate osteoporotic bone loss conditions, dioxinodehydroeckol (DHE) from edible brown alga Ecklonia cava was tested for its effects on osteoblastogenic differentiation in MC3T3-E1 pre-osteoblasts. DHE was observed to successfully enhance osteoblast differentiation, as indicated by elevated cell proliferation, alkaline phosphatase activity, intracellular cell mineralization, along with raised levels of osteoblastogenesis indicators at the concentration of 20 µM. Results suggested a possible intervening of DHE on the bone morphogenetic protein (BMP) signaling pathway, according to elevated protein levels of BMP-2, collagen-I, and Smads. In addition, the presence of DHE was also able to raise the phosphorylated extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) levels which are also activated by the BMP signaling pathway. In conclusion, DHE is suggested to be a potential bioactive compound against bone loss that could enhance osteoblastogenesis with a suggested BMP pathway interaction.

Anti-Inflammatory Activity of Heterocarpin from the Salt Marsh Plant Corydalis heterocarpa in LPS-Induced RAW 264.7 Macrophage Cells.

The inhibitory effect of three chromones 1-3 and two coumarins 4-5 on the production of nitric oxide (NO) was evaluated in LPS-induced RAW 264.7 macrophage cells. Among the compounds tested heterocarpin (1), a furochromone, significantly inhibited its production in a dose-dependent manner. In addition, heterocarpin suppressed prostaglandin E2 (PGE2) production and expression of cytokines such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß) and interleukin-6 (IL-6).

Development and Quality Characteristics of Elderly-Friendly Pulpo a La Gallega Prepared Using Texture-Modified Octopus (Octopus vulgaris) Arms.

Considering the need for developing new senior-friendly processed seafood, this study aimed to develop octopus pulpo with high preference and excellent quality characteristics for elderly individuals by optimizing the vinegar immersion and sous vide softening treatment conditions for octopus (Octopus vulgaris) arms. The optimal sous vide heating temperature (70.0-100.0 °C), time (69.5-170.5 min), and vinegar concentration (0.2-0.8%) were established using response surface methodology (RSM). The pulpo prototype was produced using an octopus arm softened under optimal conditions and seasoned with a potato and olive oil sauce. The physicochemical and nutritional properties of the prototype were evaluated, followed by sensory evaluation and safety assessments. The hardness of softened octopuses obtained by 0.48% vinegar immersion and sous vide treatment (84.3 °C, 139.8 min), determined using RSM, was 394.5 × 1000 N/m2, showing a reduction of 83.0%; this was confirmed by electron microscopic observation. The texture of the pulpo prototype with softened octopus arms showed the highest preference (8.4 points) and high physicochemical and nutritional properties. Overall, the octopus pulpo a La Gallega prototype produced using texture-modified octopus arms was suitable for consumption by elderly people with chewing disorders and could help improve their quality of life.

Development and Quality Characteristics of Jangjorim Prepared Using Long-Arm Octopus (Octopus minor) as an Elderly-Friendly Food.

We prepared a long-arm octopus Jangjorim prototype (LOJP) by optimizing the ratio of ingredients for seasoning and establishing heat sterilization parameters. The optimal amounts of purified water (2.9-56.6%, A), starch syrup (0.3-37.8%, B), and soy sauce (25.5-71.5%, C) for the production of seasoning soy sauce were obtained using response surface analysis. The LOJP was prepared by combining A, B, and C under the optimal conditions and evaluated for consumer preferences and physicochemical, nutritional, and microbiological properties and compared with Korea's legal management standards for geriatric nutrition. The hardness of the LOJP produced using the optimal mixing ratio of purified water (51.2%, 154.0 g), starch syrup (29.3%, 308.0 g), and soy sauce (19.5%, 256.9 g) was 36.7 × 1000 N/m2. This value was lower than the hardness of raw octopus (2153.6 × 1000 N/m2) by 2116.9 × 1000 N/m2. It received the highest score (8.7) in the preference evaluation of older consumers. The LOJP was classified as level 2, allowing consumption through the gums of elderly consumers per Korea's food standards for the elderly. The LOJP was the product highly preferred by elderly consumers with chewing disorders due to its ease of intake and nutritional content.

Effect and Comparison of Luteolin and Its Derivative Sodium Luteolin-4'-sulfonate on Adipogenic Differentiation of Human Bone Marrow-Derived Mesenchymal Stem Cells through AMPK-Mediated PPARγ Signaling.

Luteolin is a common phytochemical from the flavonoid family with a flavone structure. Studies reported several bioactivities for luteolin and similar flavones. Attenuating the increased adipogenesis of bone marrow cells (hBM-MSCs) has been regarded as a therapeutic target against osteoporotic bone disorders. In the present study, the potential roles of luteolin and its sulfonic acid derivative luteolin-OSO3Na in regulating adipogenic differentiation of hBM-MSCs were investigated. Adipo-induced cells were treated with or without compounds, and their effect on adipogenesis was evaluated by adipogenic marker levels such as lipid accumulation and PPARγ pathway activation. Luteolin hindered the adipogenic lipid accumulation in adipo-induced hBM-MSCs. Immunoblotting and reverse transcription-polymerase chain reaction analysis results indicated that luteolin downregulated PPARγ and downstream factors of C/EBPα and SREBP1c expression which resulted in inhibition of adipogenesis. Luteolin-OSO3Na showed similar effects; however, it was significantly less effective compared to luteolin. Investigating p38, JNK, and ERK MAPKs and AMPK activation indicated that luteolin suppressed the MAPK phosphorylation while stimulating AMPK phosphorylation. On the other hand, luteolin-OSO3Na was not able to notably affect the MAPK and AMPK activation. In conclusion, this study suggested that luteolin inhibited adipogenic differentiation of hBM-MSCs via upregulating AMPK activation. Replacing its 4'-hydroxyl group with sulfonic acid sodium salt diminished its antiadipogenic effect indicating its role in regulating AMPK activation. The general significance is that luteolin is a common phytochemical with various health-beneficial effects. The current study suggested that luteolin may serve as a lead compound for developing antiosteoporotic substances with antiadipogenic properties.

MMP-Inhibitory Effects of Flavonoid Glycosides from Edible Medicinal Halophyte Limonium tetragonum.

Limonium tetragonum has been well-known for its antioxidative properties as a halophyte. This study investigated the antimetastasis effect of solvent-partitioned L. tetragonum extracts (LTEs) and isolated compounds on HT1080 mouse melanoma cell model with a focus on matrix metalloproteinase (MMP) activity and TIMP and MAPK pathways. Upregulation and stimulation of MMPs result in elevated degradation of extracellular matrix which is part of several complications such as metastasis, cirrhosis, and arthritis. The anti-MMP capacity of LTEs was confirmed by their MMP-inhibitory effects, regulation of MMP and TIMP expression, and suppression of MAPK pathway. Among all tested LTEs, 85% aq. MeOH and n-BuOH were found to be most active fractions which later yielded two known flavonoid glycosides, myricetin 3-galactoside and quercetin 3-o-beta-galactopyranoside. Anti-MMP potential of the compounds was confirmed by their ability to regulate MMP expression through inhibited MAPK pathway activation. These results suggested that L. tetragonum might serve as a potential source of bioactive substances with effective anti-MMP properties.

Physicochemical changes in kimchi containing skate (Raja kenojei) pretreated with organic acids during fermentation.

The effects of supplementing kimchi with acetic acid (TSA)- and citric acid (TSC)-treated skate were investigated during the fermentation process. The chemical and microbiological properties of the product were also investigated. The quality parameters of kimchi were determined by measuring the pH, titratable acidity, reducing sugar content, volatile basic nitrogen (VBN), biogenic amines (BAs), and microbial analysis. During fermentation, the kimchi containing skate was observed to have a lower acidity than the control. Treatment of skate with an organic acid increased the acidity and gradually decreased the reducing sugar content during fermentation. Higher concentrations of organic acid resulted in a decrease in VBN and BAs in the TSA and TSC groups compared to those in the kimchi containing skate. The results suggest that supplementing kimchi with skate treated with organic acids affects the histamine content of the products.

Phlorotannins suppress adipogenesis in pre-adipocytes while enhancing osteoblastogenesis in pre-osteoblasts.

Osteoporosis, a prevalent bone disease in an aging population, is considered to be closely related to osteoblastogenesis and adipogenesis. As a part of an ongoing trend to develop natural substances that attenuate osteoporotic conditions, edible brown algae E. cava and its bioactive constituents were tested for their effects on adipogenic differentiation in 3T3-L1 fibroblasts and osteoblast differentiation in MC3T3-E1 pre-osteoblasts. Following an activity-based isolation, three phlorotannin derivatives, triphlorethol-A (1), eckol (2) and dieckol (3), were isolated. Anti-adipogenesis effect of phlorotannins at the concentration of 20 µM was observed by reduced lipid accumulation and the suppressed expression of adipogenic differentiation markers. In addition, isolated phlorotannins successfully enhanced the osteoblast differentiation as indicated by increased alkaline phosphatase activity along with raised levels of osteoblastogenesis indicators and intracellular calcification at the concentration of 20 µM. In conclusion, E. cava is suggested as a source for functional food ingredients, especially phlorotannin derivatives that can be utilized for extenuating osteoporosis and obesity.

Effect of Baechu Kimchi Added Ecklonia cava Extracts on High Glucose-induced Oxidative Stress in Human Umbilical Vein Endothelial Cells.

Endothelial cell dysfunction is considered to be a major cause of vascular complications in diabetes. In the present study, we investigated the protective effect of a baechu kimchi added Ecklonia cava extract (BKE) against high glucose induced oxidative damage in human umbilical vein endothelial cells (HUVECs). Treatment with a high concentration of glucose (30 mM) induced cytotoxicity, whereas treatment with BKE protected HUVECs from high glucose induced damage; by restoring cell viability. In addition, BKE reduced lipid peroxidation, intracellular reactive oxygen species and nitric oxide levels in a dose dependent manner. Treatment with high glucose concentrations also induced the overexpression of inducible nitric oxide synthase, cyclooxygenase-2 and NF-κB proteins in HUVECs, but BKE treatment significantly reduced the overexpression of these proteins. These findings indicate that BKE may be a valuable treatment against high glucose-induced oxidative stress HUVECs.

Alleviating Effects of Baechu Kimchi Added Ecklonia cava on Postprandial Hyperglycemia in Diabetic Mice.

In this study, we investigated the inhibitory effects of Baechu kimchi added Ecklonia cava on the activities of α-glucosidase and α-amylase and its alleviating effect on the postprandial hyperglycemia in STZ-induced diabetic mice. Baechu kimchi added Ecklonia cava (BKE, 15%) was fermented at 5°C for 28 days. Optimum ripened BKE was used in this study as it showed the strongest inhibitory activities on α-glucosidase and α-amylase by fermentation time among the BKEs in our previous study. The BKE was extracted with 80% methanol and the extract solution was concentrated, and then used in this study. The BKE extract showed higher inhibitory activities than Baechu kimchi extract against α-glucosidase and α-amylase. The IC50 values of the BKE extract against α-glucosidase and α-amylase were 0.58 and 0.35 mg/mL, respectively; BKE exhibited a lower α-glucosidase inhibitory activity but a higher α-amylase inhibitory activity than those of acarbose. The BKE extract alleviated postprandial hyperglycemia caused by starch loading in normal and streptozotocin-induced diabetic mice. Furthermore, the BKE extract significantly lowered the incremental area under the curve in both normal and diabetic mice (P<0.05). These results indicated that the BKE extract may delay carbohydrate digestion and thus glucose absorption.