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Highlights • Lipid accumulation in the 3T3-L1 cells were inhibited by treatment with DRE. • The expression levels of SREBP1c, PPARγ, C/EBPα, and FAS were decreased by DRE. • HFD induced fat mice showed lower rate of weight gain and serum TG level through DRE administration., This study confirms the anti-obesity effect of the ethyl acetate fraction of Distylium racemosum (DRE), a member of Hamamelidaceae, that naturally grows on Jeju Island, on adipocyte differentiation in 3T3-L1 cells. This study further demonstrated that DRE exhibits anti-obesity effects in C57BL/6 obese mice. The degree of adipocyte differentiation was determined using Oil red O stain; results indicated a decrease in fat globules, which was dependent on DRE concentration, when pre-adipocytes were treated with differentiation-inducing agents. In addition, this significantly reduced the expression of the adipogenic transcription factor and related genes. C57BL/6 obese mice treated with DRE showed a lower rate of body weight gain than the high-fat diet (HFD) group mice. Further, the level of serum triglyceride in the DRE treatment group was lower than that in the HFD group. The findings show that DRE are capable of suppressing adipocyte accumulation; therefore, DRE may represent a promising source of functional materials for the anti-obesity.

A phytochemical application of leaves from Ficus erecta var. sieboldii (Miq.) King has not been widely investigated. We determined an anti-inflammatory effect of F. erecta extracts on lipopolysaccharide (LPS)-mediated production through modulation of several pro-inflammatory mediators and prostaglandin E2 (PGE2). Among the F. erecta extracts, the CH2Cl2 fraction (CFE) most effectively suppressed the LPS-mediated production of nitric oxide (NO) in Raw264.7 cells. As determined by immunoblotting and PCR, CFE was shown to have an inhibitory effect on LPS-induced mRNA and protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). In addition, CFE showed significant inhibitory effects on LPS-mediated production of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and PGE2 (P2 production were syringaresinol (C1) and 6,7-furano-5-methoxy hydrocoumaric acid (C2). In conclusion, CFE showed an inhibitory effect on LPS-mediated inflammatory responses by suppressing iNOS, COX-2, TNF-α, IL-1β, and IL-6 production. The compounds C1 and C2 showed strong inhibitory effects on LPS-mediated production of PGE2 and may be applicable as starter compounds for developing anti-inflammatory and anti-nociceptive drugs.

3-Bromo-4,5-dihydroxybenzaldehyde (BDB) is a natural bromophenol compound that is most commonly isolated from red algae. The present study was designed to investigate the anti-inflammatory properties of BDB on atopic dermatitis (AD) in mice induced by 2,4-dinitrochlorobenzene (DNCB) and on lipopolysaccharide (LPS)-stimulated murine macrophages. BDB treatment (100 mg/kg) resulted in suppression of the development of AD symptoms compared with the control treatment (induction-only), as demonstrated by reduced immunoglobulin E levels in serum, smaller lymph nodes with reduced thickness and length, a decrease in ear edema, and reduced levels of inflammatory cell infiltration in the ears. In RAW 264.7 murine macrophages, BDB (12.5, 25, 50, and 100 μM) suppressed the production of interleukin-6, a proinflammatory cytokine, in a dose-dependent manner. BDB also had an inhibitory effect on the phosphorylation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and signal transducer and activator of transcription 1 (STAT1; Tyr 701), two major signaling molecules involved in cellular inflammation. Taken together, the results show that BDB treatment alleviates inflammatory responses in an atopic dermatitis mouse model and RAW 264.7 macrophages. These results suggest that BDB may be a useful therapeutic strategy for treating conditions involving allergic inflammation such as atopic dermatitis.

We evaluated the effects of Cynanchum wilfordii (CW) ethanolic extract on blood cholesterol levels in adults with high low-density lipoprotein cholesterol (LDL-C) levels. In a double-blind, randomized, placebo-controlled, parallel trial, 84 subjects were recruited. Participants were randomly divided into two groups with a low-dose (300 mg/d) or high-dose (600 mg/d) of CW. Levels of very low-density lipoprotein (p = 0.022) and triglycerides (p = 0.022) were significantly lower in the low-dose CW group than in the placebo group after 8 weeks. In a subgroup of participants with LDL-C&ge, 150 mg/dL (n = 33), there was a significant decrease in total cholesterol (low-dose, p = 0.012, high-dose, p = 0.021), apolipoprotein B (low-dose, p = 0.022, high-dose, p = 0.016), and cholesteryl ester transfer protein (low-dose, p = 0.037, high-dose, p = 0.016) after 8 weeks of CW. The correlation between changes in total cholesterol and baseline LDL-C levels was significant in the groups that received both doses of CW (low-dose, p = 0.010, high-dose, p = 0.015). These results show that the CW ethanolic extract can regulate blood cholesterol in subjects with LDL-C&ge, 150 mg/dL.

This study determined the anti-obesity effect of Crinum asiaticum var. japonicum Baker extract (CAE) on adipocytes and obese mice. The inhibitory effects of CAE on adipocyte differentiation and adipogenesis were determined using differentiation induction medium in 3T3-L1 cells. To get an insight into underlying molecular actions of CAE, we investigated the changes in the expression levels of genes involved in lipogenesis by CAE treatment using qRT-PCR. CAE strongly suppressed adipocyte differentiation through downregulation of PPARγ, C/EBPα, C/EBP β, and aP2. CAE treatment could also suppress the expression levels of ACC, FAS, LPL and HMGCR gene in 3T3-L1 cells. Male C57BL/6 strain and C57BL/6J-ob/ob strain mice were fed with HFD containing 60% fat and normal diet in the presence or absence of 25, 50, and 100mg/kg CAE for 7 weeks. CAE supplementation could highly suppress the body weight gain and epididymal fat accumulation without changes in food uptake in both obese models. Increases in total cholesterol, LDL-cholesterol and triglyceride were highly suppressed in the presence of CAE. In summary, CAE has an anti-obesity effect and this anti-obesity potential might be associated with downregulation of genes involved in adipocyte differentiation and lipogenesis.

Allergic skin inflammation such as atopic dermatitis is characterized by skin barrier dysfunction, edema, and infiltration with various inflammatory cells. The anti-inflammatory effects of Apo-9'-fucoxanthinone, isolated from Sargassum muticum, have been described in many diseases, but the mechanism by which it modulates the immune system is poorly understood. In this study, the ability of Apo-9'-fucoxanthinone to suppress allergic reactions was investigated using a mouse model of atopic dermatitis. The Apo-9'-fucoxanthinone-treated group showed significantly decreased immunoglobulin E in serum. Also, Apo-9'-fucoxanthinone treatment resulted in a smaller lymph node size with reduced the thickness and length compared to the induction group. In addition, Apo-9'-fucoxanthinone inhibited the expression of interleukin-4, interferon-gamma and tumor necrosis factor-alpha by phorbol 12-myristate 13-acetate and ionomycin-stimulated lymphocytes. These results suggest that Apo-9'-fucoxanthinone may be a useful therapeutic strategy for treating chronic inflammatory diseases.

The anti-inflammatory activities of the products of supercritical fluid extraction of Litsea japonica fruit (SFELJF) using supercritical carbon dioxide were investigated. The SFELJF extract dose-dependently inhibited the production of inflammatory markers [nitric oxide (NO), inducible nitric oxide synthase (iNOS), prostaglandin E2 (PGE2), and cyclooxygenase-2 (COX-2)] and proinflammatory cytokines [tumour necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-6] induced by lipopolysaccharide (LPS) treatment. To further elucidate the mechanisms underlying these inhibitory effects, LPS-induced nuclear factor-κB (NF-κB) activation and mitogen-activated protein kinase (MAPK) phosphorylation were studied. The SFELJF extract inhibited NF-κB (p65 and p50) activation and MAPK [extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38] phosphorylation in a dose-dependent manner. These results suggest that the anti-inflammatory activities of SFELJF extracts are due to proinflammatory cytokines and mediators via suppression of NF-κB activation and MAPK phosphorylation.

Pinus thunbergii is a black pine that has been widely used in health-promoting foods. This study was conducted to investigate the hepatoprotective effects of the alcohol-aqueous extract of Pinus thunbergii PARL leaves (PTE) in ethanol-induced liver-damaged mice. PTE increased the weight and survival rate of mice while reducing the levels of serum hepatic marker enzymes. PTE also restored the levels of antioxidant enzymes in liver tissues while drastically increasing lipid peroxidation concentrations. Histopathological examination of PTE-treated mice presented relatively less lipid droplets and hepatic tissue damage compared to ethanol-treated mice. Finally, to identify PTE components, Natural Products Application Solution with UNIFI along with an in-house library was processed and results revealed 35 compounds in the extract. These compounds were reported to exhibit significant antioxidant and/or hepatoprotective properties. Our results suggest that PTE improves liver function and can be developed as a food product with hepatoprotective effects.

The present study aimed to investigate comparative anti-inflammatory effects of Litsea japonica fruit and leaf extract considering the balance of safety and efficacy. Dose response studies were performed to determine the inhibitory effects of 70% EtOH extract of leaf (L70%) on the pro-inflammatory enzymes expression, COX-2/PGE2 and NO/iNOS, and pro-inflammatory cytokines production, IL-1β, IL-6, and TNF-α in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. We also examined comparative effects of 30 and 70% EtOH extract of fruits (F30% and F70%) at low concentration (10 ㎍/㎖) in the same conditions. L70% at 50 and 100 ㎍/㎖ showed inhibitory effects on almost all the inflammatory mediators we examined except for COX-2 regulation, but there were no effects at 10 ㎍/㎖. Since 100 ㎍/㎖ of L70% have 18.2% cytotoxicity, we compared the effects of fruit extract, F30% and F70% at 10 ㎍/㎖ on the regulation of NO/iNOS, PGE2, IL-1β, IL-6, and TNF-α and obtained that fruit extacts are more efficacious and safe than leaf. This study suggests that the 30% EtOH fraction of L. japonica fruit could be a good candidate for development as a functional food supplement in the prevention of inflammatory disorders.

The anti-inflammatory effects of compounds isolated from the Litsea japonica fruit were investigated in LPS-stimulated murine macrophage (RAW 264.7) cells. The results indicated that litsenolide (LN)B2 significantly inhibited the LPS-induced release of pro-inflammatory mediators, such as NO and PGE 2 . LNC2, and, on the other hand, exhibited cytotoxicity at the same concentrations at which it exhibited an inhibitory property. Therefore, only LNB2 was used for further experimentation. LNB2 reduced the LPS-induced production of pro-inflammatory cytokines. We further investigated the mechanism by which LNB2 inhibits pro-inflammatory mediators and cytokines by examining the level of NF-κB and MAPKs phosphorylation, which all serve as key components in inflammation-induced signaling pathways in RAW 264.7 cells. LNB2 inhibited the LPS-induced phosphorylation of NF-κB and MAPKs. These results suggest that LNB2 has an anti-inflammatory effect, inhibiting the production of pro-inflammatory mediators and cytokines via inhibition of NF-κB and MAPK signaling in LPS-stimulated RAW 264.7 cells.

This study describes a preliminary evaluation of the anti-inflammatory activity of Castanopsis cuspidata extracts. C. cuspidata was extracted using 80% ethanol and then fractionated sequentially with n -hexane, dichloromethane, ethylacetate, and butanol. To screen for anti-inflammatory agents effectively, we first examined the inhibitory effect of the C. cuspidata extracts on the production of pro-inflammatory factors and cytokines stimulated with lipopolysaccharide. In addition, we examined the inhibitory effect of C. cuspidata extracts on pro-inflammatory mediators (NO, iNOS, COX-2) in murine macrophage RAW 264.7 cells. The amounts of protein levels were determined by immunoblotting. Of the sequential solvent fractions of C. cuspidata , the n ‑hexane, dichloromethane and ethylacetate fractions inhibited the mRNA expression of pro-inflammatory cytokines (IL-1 β and IL-6), production of NO, and the protein level of iNOS and COX-2. These results suggest that C. cuspidata may have significant effects on inflammatory factors and may be provided as a possible anti-inflammatory therapeutic plant.

Fruits of the Litsea family of trees and shrubs contain biologically active compounds, some of which have been used as natural nutrients and flavoring agents in food. In this study, we identified novel anti-nociceptive effects of the 30% ethanol extract, the CH(2)Cl(2) fraction and the associated active components (Hamabiwalactone A and B) from Litsea japonica fruit by using in vivo peripheral and central nervous pain models. In addition, we compared the anti-inflammatory effects of several fractions from L. japonica fruit extracts using lipopolysaccharide (LPS)-stimulated Raw264.7 cells. The CH(2)Cl(2) fraction of L. japonica fruit (LJM) had an optimal combination of anti-inflammatory effects and low cytotoxicity. Dose response studies were performed to determine the inhibitory effects of LJM on the pro-inflammatory enzymes, COX-2/PGE(2) and NO/iNOS, and pro-inflammatory cytokines, IL-1β, IL-6, and TNF-α. Molecular profiling revealed that LJM exerts anti-inflammatory effects through inhibition of NF-κB and JNK/p38 MAPK signaling in LPS-induced macrophages. This study suggests that CH2Cl2 fraction of L. japonica fruit and its bioactive components are potential candidates as anti-inflammatory and analgesic agents (painkillers) for the treatment of inflammatory diseases.

We investigated the composition of essential oil from fingered citron (Citrus medica L. var. sarcodactylis) (FCEO) peels by GC–MS and its anti-inflammatory effects on lipopolysaccharide (LPS) – stimulated mouse macrophage (RAW 264.7) cells. Fifteen compounds, representing 98.97% of the essential oil, were tentatively identified; the main constituents were limonene (52.44%) and γ-terpinene (28.41%). FCEO significantly inhibited nitric oxide (NO) and prostaglandin E2 (PGE2) by suppressing the protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2, respectively. Additionally, FCEO suppressed the production of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-6. FCEO attenuated LPS-induced nuclear factor-κB (NF-κB) activation via inhibition of inhibitor κB-α phosphorylation. Furthermore, FCEO blocked activation of c-Jun N-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK) but not that of p38 mitogen-activated protein kinase. These results indicate that FCEO inhibits LPS-stimulated inflammation by blocking the NF-κB, JNK, and ERK pathways in macrophages, and demonstrate that FCEO possesses anti-inflammatory properties.

Inflammatory cytokines play a major role in osteoclastogenesis, leading to the bone resorption that is frequently associated with osteoporosis. Sargachromanol G (SG), isolated from the brown alga Sargassum siliquastrum, inhibits the production of inflammatory cytokines. In the present study, we determined the effect of SG on receptor activator of NF-κB ligand (RANKL)-induced osteoclast formation. SG inhibited RANKL-induced osteoclast differentiation from RAW264.7 cells without signs of cytotoxicity. Additionally, the expression of osteoclastic marker genes, such as tartrate-resistant acid phosphatase (TRAP), cathepsin K (CTSK), matrix metalloproteinase 9 (MMP9), and calcitonin receptor (CTR), was strongly inhibited. SG inhibited RANKL-induced activation of NF-κB by suppressing RANKL-mediated IκB-α degradation. Furthermore, SG inhibited RANKL-induced phosphorylation of mitogen activated protein kinases (p38, JNK, and ERK). This study identified SG as an inhibitor for osteoclast formation and provided evidence that natural compounds, such as SG, are an alternative medicines for preventing and treating osteolysis.

The cytoprotective effect of quercitrin (QR) against oxidative stress induced cell damage by hydrogen peroxide (H2O2) in Chinese hamster lung fibroblast (V79-4) cells was investigated. QR evidenced a scavenging effect of 1,1-diphenyl-2-picrylhydrazyl (DPPH), superoxide, hydroxyl radicals and on intracellular ROS, and thus prevented lipid peroxidation. As a result, QR reduced H2O2-induced cell death and apoptosis in V79-4 cells. Moreover, H2O2 induced the cleavage of caspase-3, -9, and poly-ADP-ribose polymerase (PARP) and a reduction in Bcl-xL levels, whereas pretreatment with QR significantly inhibited caspase-3, -9, and PARP cleavage and the reduction in Bcl-xL levels, and ultimately ameliorated H2O2-induced apoptosis. Taken together, these results indicate that the treatment of V79-4 cells with QR can block H2O2-induced apoptosis via the regulation of Bcl-xL. QR may be exploited as a biopreservative in food applications or as a health supplement to alleviate oxidative stress.

The aim of this study was to investigate the cytoprotective properties of the ethyl acetate fraction of Sargassum muticum (SME) against ultraviolet B (UVB)-induced cell damage in human keratinocytes (HaCaT cells). SME exhibited scavenging activity toward the 1,1-diphenyl-2-picrylhydrazyl radicals and hydrogen peroxide (H(2)O(2)) and UVB-induced intracellular reactive oxygen species (ROS). SME also scavenged the hydroxyl radicals generated by the Fenton reaction (FeSO(4) + H(2)O(2)), which was detected using electron spin resonance spectrometry. In addition, SME decreased the level of lipid peroxidation that was increased by UVB radiation, and restored the level of protein expression and the activities of antioxidant enzymes that were decreased by UVB radiation. Furthermore, SME reduced UVB-induced apoptosis as shown by decreased DNA fragmentation and numbers of apoptotic bodies. These results suggest that SME protects human keratinocytes against UVB-induced oxidative stress by enhancing antioxidant activity in cells, thereby inhibiting apoptosis.

In this study, the potent anti-tumor effects of brown algae on human leukemia HL-60 cells were investigated. The Sargassum siliquastrum extract among the 14 species of brown algae exhibited profound growth inhibitory effect on HL-60 cells in the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, therefore, S. siliquastrum was selected for use in further experiments. The highest inhibitory activity of S. siliquastrum on HL-60 cells was detected in the chloroform fraction, and the active compound was identified as a kind of chromene, sargachromanol E (SE). SE treatment showed significant growth inhibitory effects on HL-60 cells in a dose-dependent manner by inducing apoptosis, as evidenced by the formation of apoptotic bodies, fragmented DNA ladder, and the accumulation of DNA in the sub-G1 phase of cell cycle. SE induced apoptosis was accompanied by downregulation of Bcl-xL, upregulation of Bax, activation of caspase-3, and cleavage of poly (ADP-ribose) polymerase (PARP). Moreover, z-DEVD-fmk, a caspase-3 inhibitor, significantly inhibited cell cytotoxicity, apoptotic characteristics such as apoptotic bodies, sub-G1 DNA content, and cleavage of PARP induced by SE. These results suggest that SE exerts its growth inhibitory effects on HL-60 cells through caspase-3-mediated induction of apoptosis. Therefore, SE offers promising chemotherapeuric potential to prevent cancers such as human leukemia.

Melanogenesis is a well-known physiological response of human skin that may occur because of exposure to ultraviolet light, for genetic reasons, or due to other causes. In our efforts to find new skin lightening agents, palmitoleic acid was investigated for its ability to inhibit melanogenesis. In this study, palmitoleic acid's effect on melanin formation was assessed. Results indicated that palmitoleic acid was shown to down-regulate melanin content in a dose-dependent pattern. To clarify the target of palmitoleic acid action in melanogenesis, we performed Western blotting for tyrosinase, tyrosinase-related protein-1 (TRP-1), TRP-2, and microphthalmia-associated transcription factor (MITF), which are key melanogenic enzymes. Palmitoleic acid inhibited tyrosinase, TRP-2, and MITF expressions in a dose-dependent manner. However, it did not inhibit TRP-1 expression. In order to assess its usefulness in future cosmetic product applications, the cytotoxic effects of the palmitoleic acid were also determined by colourimetric MTT assays using human keratinocyte HaCaT cells. Palmitoleic acid exhibited no cytotoxicity at 500 muM in a human cell line. Therefore, this study suggests that palmitoleic acid is a candidate anti-melanogenic agent, and it might be effective in hyperpigmentation disorders.

Despite its beneficial role in host defense mechanisms, excessive nitric oxide (NO) production by activated macrophages has been implicated in several inflammatory diseases. To clarify the mechanisms of the anti-inflammatory activities of Sargassum micracanthum, we evaluated whether extracts of S. micracanthum could modulate the production of NO by activated macrophages. S. micracanthum were extracted with 80% EtOH. The extract was then successively partitioned with hexane, CH 2 Cl 2 , EtOAc, BuOH, and water. The results indicate that the hexane and CH2Cl2 fractions of S. micracanthum extract were effective inhibitors of LPS-induced NO and prostaglandin E2 (PGE2) production in RAW 264.7 cells. The inhibitory effects of the hexane and CH2Cl2 fractions of S. micracanthum were accompanied by dosedependent decreases in the production of iNOS and COX-2 proteins and iNOS and COX-2 mRNA expression. To test the inhibitory effects of S. micracanthum fractions on other cytokines, we also performed ELISA and RT-PCR assays for TNF- , IL-1s, and IL-6 in LPSstimulated RAW 264.7 macrophage cells. In these assays, the hexane and CH2Cl2 fractions of S. micracanthum produced dose-dependent decreases in the production and mRNA expression of TNF- , IL-1s, and IL-6. To test the potential application of S. micracanthum extract as a cosmetic material, we also performed MTT assays on human dermal fibroblast cells, as well as primary skin irritation tests. In these assays, S. micracanthum extracts did not induce any adverse reactions. Based on these results, we suggest that S. micracanthum extracts may be considered potential anti-inflammatory candidates for topical application.

We elucidated the pharmacological and biological effects of Oenothera laciniata extracts on the production of inflammatory mediators in macrophages. The CH(2)Cl(2) fraction of O. laciniata extract effectively inhibited LPS-induced NO, PGE(2), and proinflammatory cytokine production in RAW264.7 cells. These inhibitory effects of the CH(2)Cl(2) fraction of O. laciniata were accompanied by decreases in the expression of iNOS and COX-2 proteins and iNOS, COX-2, TNF-alpha, IL-1beta, and IL-6 mRNA. Asiatic acid and quercetin were present in the HPLC fingerprint of the O. laciniata extract. We tested the potential application of O. laciniata extract as a cosmetic material by performing primary skin irritation tests. In New Zealand white rabbits, primary irritation tests revealed that application of O. laciniata extracts (1%) did not induce erythema or edema formation. Human skin primary irritation tests were performed on the normal skin (upper back) of 30 volunteers to determine if any material in O. laciniata extracts had irritation or sensitization potential. In these assays, O. laciniata extracts did not induce any adverse reactions. Based on these results, we suggest that O. laciniata extracts be considered possible anti-inflammatory candidates for topical application.

Gracilaria verrucosa is a common marine red alga that has anti-oxidant and anti-cancer properties. Recently, we reported that anti-inflammatory constituents of G. verrucosa operate through an unknown mechanism. For this reason, we isolated two enone fatty acids from G. verrucosa and investigated their molecular mechanism in LPS-stimulated RAW264.7 cells. We found that the two compounds inhibited the production of inflammatory markers (nitric oxide, TNF-alpha, and IL-6) in a dose-dependent manner. We next studied the effects of G. verrucosa compounds on LPS-induced signaling pathways. The two compounds suppressed NF-kappaB reporter activity by interfering with nuclear translocation of NF-kappaB and suppressed JAK/STAT (p-STAT1) signaling. These results suggest that G. verrucosa inhibits the production of inflammatory mediators (NO, TNF-alpha, and IL-6) by suppressing the activation of NF-kappaB and the phosphorylation of STAT1.

This study was designed to analyze the chemical composition of Citrus obovoides (Geumgamja) and Citrus natsudaidai (Cheonyahagyul) oils and to test their biological activities. These citrus essential oils were obtained by steam distillation of fruits collected from Jeju Island, Korea, and were analyzed using gas chromatograph (GC)-flame ionization detectors (FID) and GC-MS. Limonene and gamma-terpinene were the major components of the two citrus species. To evaluate in vitro anti-acne activity, they were tested against Propionibacterium acnes and Staphylococcus epidermidis, which are involved in acne. The Geumgamja and Cheonyahagyul oils exhibited antibacterial activity against both P. acnes and S. epidermidis. Their effects on DPPH radical scavenging, superoxide anion radical scavenging, and nitric oxide radical were also assessed. Cheonyahagyul and Geumgamja exhibited only superoxide anion radical-scavenging activity. To assess their potential usefulness in future cosmetic product applications, the cytotoxic effects of the two oils were determined by colorimetric MTT assays using two animal cell lines: normal human fibroblasts and HaCaT cells. They exhibited low cytotoxicity at 0.1 microl/ml in both cell lines. In addition, they reduced P. acnes-induced secretion of interleukin-8 (IL-8) and tumor necrosis factor alpha (TNF-alpha) in THP-1 cells, an indication of anti-inflammatory effects. Therefore, based on these results, we suggest that Geumgamja and Cheonyahagyul essential oils are attractive acne-mitigating candidates for topical application.

The solvent extracts of Sargassum muticum, which were extracted by using several solvents with different polarities, were prepared for use as natural preservatives. The S. muticum extract with 80% ethanol was sequentially fractionated with n.hexane, dichloromethane, ethylacetate, and butanol. In order to effectively screen for natural preservatives agents, we first investigated the antioxidant activities such as DPPH radical scavenging capacity, superoxide radical scavenging capacity, and xanthine oxidase inhibitory activity of the S. muticum extracts. Through the screening system, we found that dichloromethane and ethylacetate fraction had high antioxidant activity with increments of the extract concentration. The antimicrobial activities and cell growth inhibition were investigated for each strain with the different concentrations of S. muticum extracts. Antimicrobial activities were shown in ethanol, dichloromethane, and n.hexane fractions of S. muticum. However, butanol, ethylacetate and water fractions showed weak antimicrobial activity against the tested microorganisms. Among the five fractions, dichloromethane fraction showed the highest antimicrobial activities against microorganisms tested, such as Bacillus sublitis, Listeria monocytogenes, Staphylococcus aureus, Escherichia coli, Salmonella Enteritidis and Pseudomonas aeruginosa. The polyphenolic compounds from ethanol, n.hexane, dichloromethane, ethylacetate, butanol, and water fractions were 63.96 mg/g, 8.49 mg/g, 28.11 mg/g, 172.64 mg/g, 114.56 mg/g, and 34.91 mg/g, respectively. The dichloromethane fraction could be suitable for development as a food preservative.

The anti-inflammatory activity of Eurya emarginata (Thumb) Makino, of which leaves have been traditionally used to treat ulcers or diuretic in Jeju Island, has been investigated in the present study. Through the phytochemical study from the methanol extract of E. emaginiata, eutigosides B and C were isolated as the active components. Sseveral inflammatory markers including TNF-alpha, IL-1beta, IL-6, NO, iNOS, and COX-2 were examined. Eutigosides B and C potentially inhibited production of pro-inflammatory cytokines (IL-6 and TNF-alpha) in a dose-dependent manner. Additionally, the intracellular contents of iNOS protein were markedly decreased after treatment with eutigosides B and C. The inhibition of iNOS activity was correlated with the decrease in nitrite levels. These results suggest that eutigoside B and C from E. emarginata may have anti-inflammatory activity through the inhibition of pro-inflammatory cytokines (TNF-alpha and IL-6), iNOS and COX-2.

Inflammation is complex process involving a variety of immune cells that defend the body from harmful stimuli. However, pro-inflammatory cytokines and inflammatory mediators can also exacerbate diseases such as cancer. The aim of this study was to identify a natural effective remedy for inflammation. We isolated a functional algal chromene compound from Sargassum siliquastrum, named sargachromanol D (SD). We evaluated the anti-inflammatory effect of SD on lipopolysaccharide (LPS)-exposed RAW 264.7 cells by measuring cell viability, cytotoxicity, and production of inflammatory mediators such as nitric oxide (NO), prostaglandin E2 (PGE2), inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), and the pro-inflammatory cytokines tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-6. SD inhibited production of NO and PGE2 from LPS-induced cells by preventing the expression of inflammatory mediators such as iNOS and COX-2 in a dose-dependent manner. Concurrently, levels of the pro-inflammatory cytokines TNF-α, IL-1β, and IL-6 were reduced with increasing concentrations of SD. In addition, SD inhibited the activation of NF-κB and mitogen-activated protein kinases (MAPKs) pathways in a concentration-dependent manner. These results indicate that SD inhibits LPS-stimulated inflammation by inhibition of the NF-κB and MAPKs pathways in macrophages.

To assess the potential therapeutic properties of trans-1,3-diphenyl-2,3-epoxypropane-1-one (DPEP), its anti-inflammatory effects were investigated in lipopolysaccharide (LPS)-stimulated mouse macrophage (RAW 264.7) cells. DPEP induced dose-dependent reduction of the protein levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) and concomitant reduction in the production of NO and prostaglandin E(2) (PGE(2)). Additionally, DPEP suppressed the production of inflammatory cytokines, including tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-6. We investigated the mechanism by which DPEP inhibits NO and PGE(2) by examining the level of nuclear factor-κB (NF-κB) activation within the mitogen-activated protein kinase (MAPK) pathway, which is an inflammation-induced signaling pathway in RAW 264.7 cells. DPEP inhibited LPS-induced phosphorylation of ERK, JNK, and p38. Furthermore, DPEP inhibited the LPS-induced phosphorylation of inhibitor κB (IκB)-α and NF-κB p50. Taken together, the results of this study demonstrate that DPEP inhibits LPS-stimulated inflammation by blocking the NF-κB and MAPK pathways in macrophages.

Nitric oxide (NO) is a crucial molecule in inflammatory diseases and is synthesized from L-arginine by a specific enzyme, NO synthase (NOS). The expression of inducible NOS (iNOS) is activated in macrophages by various stimuli, such as lipopolysaccharide (LPS), a wall component of gram-negative bacteria. LPS binds to toll-like receptor 4 (TLR4) on the macrophage surface and activates several downstream signaling pathways, including mitogen-activated protein kinase (MAPK) and nuclear factor (NF)-κB pathways. This study investigated whether sargaquinoic acid isolated from Sargassum siliquastrum might have anti-inflammatory activity and interfere with NO production in macrophages by disrupting LPS-induced signaling. This study was conducted in vitro using RAW264.7 murine macrophages. LPS-stimulated cells were treated with sargaquinoic acid, and the effects on NO production, iNOS expression, and involvement of the NF-κB signaling pathway were investigated by Griess assay, western blotting, and confocal microscopy. The results demonstrated that sargaquinoic acid inhibited the production of NO and the expression of the iNOS protein in LPS-stimulated RAW264.7 macrophages. Moreover, sargaquinoic acid inhibited the degradation of inhibitory-κB protein (IκB)-α and the nuclear translocation of NF-κB, a key transcription factor for the regulation of iNOS expression. Also, sargaquinoic acid influenced the phosphorylation of JNK1/2 MAPK, except ERK1/2 and p38 MAPKs, stimulated by LPS. These results suggest that sargaquinoic acid specifically prevents NO production in macrophages via the blockade of NF-κB activation and may thus have therapeutic applications in various inflammatory diseases.

A study on the anti-inflammatory activity of brown alga Sargassum siliquastrum led to the isolation of sargachromanol G (SG). In this study, the anti-inflammatory effect and the action mechanism of SG have been investigated in murine macrophage cell line RAW 264.7. SG dosedependently inhibited the production of inflammatory markers [nitric oxide (NO), inducible nitric oxide synthase (iNOS), prostaglandin E(2) (PGE(2)), and cyclooxygenase-2 (COX-2)] and pro-inflammatory cytokines [tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-6] induced by LPS treatment. To further elucidate the mechanism of this inhibitory effect of SG, we studied LPS-induced nuclear factor-κB (NF-κB) activation and mitogen-activated protein kinases (MAPKs) phosphorylation. SG inhibited the phosphorylation IκB-α and NF-κB (p65 and p50) and MAPK (ERK1/2, JNK, and p38) in a dose dependent manner. These results suggest that the anti-inflammatory activity of SG results from its modulation of pro-inflammatory cytokines and mediators via the suppression of NF-κB activation and MAPK phosphorylation.

In this study, the anti-inflammatory effect of fucoxanthin (FX) derivatives, which was isolated from Sargassum siliquastrum were evaluated by examining their inhibitory effects on pro-inflammatory mediators in lipopolysaccharide (LPS)-stimulated murine macrophage RAW 264.7 cells. The FX derivatives were isolated from activity-guided chloroform fraction using inhibition of nitric oxide (NO) production and identified as 9'-cis-(6'R) fucoxnathin (FXA), and 13-cis and 13'-cis-(6'R) fucoxanthin complex (FXB) on the basis of a comparison of NMR spectroscopic data. Both FXA and FXB significantly inhibited the NO production and showed slightly reduce the PGE2 production. However, FXB exhibited cytotoxicity at the whole tested concentration, therefore, the results of FXA was only illustrate for further experiments. FXA induced dose-dependent reduction in the inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) proteins as well as mRNA expression. In addition, FXA reduced the LPS-stimulated production and mRNA expressions of TNF-α and IL-6 in a dose-dependent manner whereas IL-1β production do not inhibit by addition of FXA. Taken together, these findings indicate that the anti-inflammatory properties of FXA may be due to the inhibition of iNOS/NO pathway which associated with the attenuation of TNF-α and IL-6 formation. Thus FXA may provide a potential therapeutic approach for inflammation related diseases.

Bone diseases are characterized by the presence of pro-inflammatory cytokines that regulate bone turnover. The receptor activator of NF-κB ligand (RANKL) is a soluble osteoblast-derived protein that induces bone resorption through osteoclast differentiation and activation. Sargachromanol G (SG) was isolated from the brown algae Sargassum siliquastrum; SG has anti-osteoclastogenic activity, but its mechanism of action and its active components remain largely unknown. In the present study, we investigated the anti-osteoclastogenic effects of SG on the expression of interleukin-1β (IL-1β)-induced osteoclastogenic factors (PGE(2), COX-2, IL-6, OPG, and RANKL) in the human osteoblast cell line MG-63. We also examined the role of the nuclear factor-κB (NF-κB) and the mitogen-activated protein kinase (MAPK) signaling pathways in IL-1β-stimulated MG-63 cells. SG dose-dependently inhibited the production of osteoclastogenic factors in MG-63 cells. SG also inhibited phosphorylation of MAPK (ERK1/2, p38, and JNK) and NF-κB (p65, p50, and IκB-α). These results suggest that the anti-osteoporotic effect of SG may be because of the modulation of osteoclastogenic factors via suppression of MAPK and NF-κB activation.

It has been previously determined that pro-inflammatory mediators including nitric oxide (NO), prostaglandin E 2 (PGE 2 ), tumor necrosis factor (TNF)-α, and interleukin (IL)-1β and IL-6 contribute to the courses of a variety of inflammatory diseases. In this study, we evaluated the anti-inflammatory effects of fucoxanthin (FX), a natural biologically active substance isolated from Ishige okamurae , by determining its inhibitory effects on pro-inflammatory mediators in lipopolysaccharide (LPS)-stimulated murine macrophage RAW 264.7 cells. FX induced dose-dependent reductions in the levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) proteins and concomitant reductions in the production of NO and PGE 2 . Additionally, FX was shown to suppress the production of inflammatory cytokines including IL-1β, TNF-α, and IL-6. Furthermore, FX inhibited the cytoplasmic degradation of inhibitors of B (IκB)-α and the nuclear translocation of p50 and p65 proteins, resulting in lower levels of nuclear factor (NF)-κB transactivation. Additionally, FX was shown to induce a dose-dependent inhibition of the phosphorylation of mitogen-activated protein kinases (MAPKs; JNK, ERK and p38). Collectively, the results of this study demonstrate that FX reduces the levels of pro-inflammatory mediators including NO, PGE 2 , IL-1β, TNF-α, and IL-6 via the inhibition of NF-κB activation and the suppression of MAPK phosphorylation in RAW 264.7 cells. These findings reveal, in part, the molecular basis underlying the anti-inflammatory properties of FX.

Cinnamomum camphora Sieb (Lauraceae) has long been prescribed in traditional medicine for the treatment of inflammation-related diseases such as rheumatism, sprains, bronchitis and muscle pains. In this study, therefore, we aimed to investigate the inhibitory effects of Cinnamomum camphora on various inflammatory phenomena to explore its potential anti-inflammatory mechanisms under non-cytotoxic (less than 100 microg/ml) conditions. The total crude extract (100 microg/ml) prepared with 80% methanol (MeOH extract) and its fractions (100 microg/ml) obtained by solvent partition with hexane and ethyl acetate (EtOAc) significantly blocked the production of interleukin (IL)-1 beta, IL-6 and the tumor necrosis factor (TNF)-alpha from RAW264.7 cells stimulated by lipopolysaccharide (LPS) up to 20-70%. The hexane and EtOAc extracts (100 microg/ml) also inhibited nitric oxide (NO) production in LPS/interferon (IFN)-gamma-activated macrophages by 65%. The MeOH extract (100 microg/ml) as well as two fractions (100 microg/ml) prepared by solvent partition with n-butanol (BuOH) and EtOAc strongly suppressed the prostaglandin E(2) (PGE(2)) production in LPS/IFN-gamma-activated macrophages up to 70%. It is interesting to note that hexane, BuOH and EtOAc extracts (100 microg/ml) also inhibited the functional activation of beta1-integrins (CD29) assessed by U937 homotypic aggregation up to 70-80%. Furthermore, EtOAc and BuOH extracts displayed strong anti-oxidative activity with IC(50) values of 14 and 15 microM, respectively, when tested by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) and xanthine oxide (XO) assays. Taken together, these data suggest that the anti-inflammatory actions of Cinnamomum camphora may be due to the modulation of cytokine, NO and PGE(2) production and oxidative stress, and of the subfractions tested, the EtOAc extract may be further studied to isolate the active anti-inflammatory principles.

The chemical composition and antiinflammatory activities of hydrodistilled essential oil from Neolitsea sericea leaves (NSE) have been investigated for the first time. The chemical constituents of NSE were analysed by GC-MS and found to include sericenine (32.3%), sabinene (21.0%), trans-β-ocimene (13.3%), β-caryophyllene (4.8%), and 4-terpineol (4.2%). The effects of NSE on nitric oxide (NO), prostaglandin E2 (PGE2), tumour necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 production in lipopolysaccharide (LPS)-activated RAW 264.7 macrophages were also examined. Pro-inflammatory cytokine and mediator tests indicated that NSE has excellent dose-dependent inhibitory activities. To further examine the mechanism responsible for the inhibition of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression by NSE, we examined the effect of NSE on nuclear factor-κB (NF-κB) activation and the phosphorylation of mitogen-activated protein kinases (MAPK). NSE inhibited NF-κB activation by LPS, and this was associated with the abrogation of IκB-α phosphorylation and subsequent decreases in nuclear p50 and p65 protein levels. Further, the phosphorylation of p38, ERK and JNK was suppressed by NSE in a concentration-dependent manner. These results suggest that NSE exerts antiinflammatory effects in LPS-stimulated RAW 264.7 macrophages by inhibition of NF-κB activation and MAPK phosphorylation, and, therefore, may be useful for treatment of inflammatory diseases.