Alpha-Linolenic Acid Alleviates Dextran Sulfate Sodium-Induced Ulcerative Colitis in Mice.

Ulcerative colitis (UC) is a type of inflammatory bowel disease characterized by inflammation of the large intestine, rectal bleeding, and abdominal pain. It can be alleviated by certain bioactive compounds, including α-linolenic acid (ALA), which is a bioactive component in fermented black radish (Raphanus sativus L. var. niger). The aim of this study was to evaluate the anti-inflammatory effects of ALA in dextran sulfate sodium (DSS)-induced UC in mice. UC was induced in C57BL/6 mice by allowing them to freely drink water containing 2.5% DSS for 7 days, followed by oral administration of ALA (30 and 60 mg/kg/day) or vehicle control for 7 days. DSS-induced colitis was evaluated using the Disease Activity Index (DAI) and by measuring colon length and performing a histopathological examination. Compared to the control group, the vehicle-treated group showed a higher DAI score, shorter colon, goblet cell loss, and crypt shortening. The ALA treatment mitigated clinical signs of UC and histopathological changes. Furthermore, it mitigated intestinal inflammation by reducing the expression of ionized calcium binding adaptor molecule 1-positive macrophages in the colon. These results show that ALA alleviates DSS-induced UC by suppressing colon damage, which includes goblet cell loss, crypt shortening, and a reduction of macrophages in the colon.

Fermented black radish (Raphanus sativus L. var. niger) attenuates methionine and choline deficient diet-induced nonalcoholic fatty liver disease in mice.

As one of the wide-ranging form of chronic liver disease, there are only limited therapeutic options for nonalcoholic fatty liver disease (NAFLD). We evaluated whether fermented black radish (Raphanus sativus L. var. niger; FBR) ameliorates lipid accumulation, inflammation, and hepatic fibrosis, which are characteristics of the pathogenesis of NAFLD. Fermented black radish treatment reduced lipid accumulation in 3T3-L1 adipocytes, which appeared to be associated with the downregulation of adipogenic transcription factors, including sterol regulatory element-binding protein 1c, CCAAT/enhancer-binding protein α, peroxisome proliferator-activated receptor γ, and lipid accumulation-related genes including adipocyte protein-2 and fatty acid synthase. Administration of FBR to C57BL/6J mice challenged with methionine and choline deficient (MCD) diet significantly attenuated the increased serum levels of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and triglyceride. In addition, treatment with FBR interestingly repressed the hepatic inflammation induced with MCD diet, by lowering the expression of inducible nitric oxide synthase and suppressing the inactivation of macrophages and Kupffer cells in the liver. Fermented black radish was also shown to mitigate liver fibrosis through the inhibition of alpha-smooth muscle actin, transforming growth factor beta-1, and collagen type I alpha 1 chain. Our results indicate that FBR ameliorates NAFLD and its related metabolic disease by regulating multiple pathways, suggesting that FBR may be an effective dietary supplement for ameliorating NAFLD.

Black Radish (Raphanus sativus L. var. niger) Extract Mediates Its Hepatoprotective Effect on Carbon Tetrachloride-Induced Hepatic Injury by Attenuating Oxidative Stress.

Nonalcoholic fatty liver disease is a serious liver disorder associated with oxidative stress. Black radish (Raphanus sativus L. var. niger) extract (BRE) can lower the risk of this disease. The hepatoprotective effect of BRE containing 3-(E)-(methylthio)methylene-2-pyrrolidinethione was evaluated in human hepatocyte carcinoma (HepG2) cells and in rat livers with carbon tetrachloride (CCl4)-induced hepatic injury. BRE was administered at 125, 250, 500, and 1000 µg/mL to the oleic acid-induced HepG2 cells. Male Sprague-Dawley rats were randomly divided into seven groups: the control group, BRE group, CCl4 group, and BRE + CCl4 group. BRE was administered orally at 125, 250, 500, and 1000 mg/kg/day once daily for 7 consecutive days, followed by a single oral treatment of 1.5 mL/kg CCl4. Inhibition of lipid accumulation, serum markers of liver injury, histological evaluations, levels of oxidative stress related enzymatic and nonenzymatic antioxidants in HepG2 cells and liver tissue were investigated. The protein expression of main liver P450 isoenzymes such as cytochrome p450(CYP)2E1, the expression of nuclear factor erythroid 2-related factor-2(Nrf-2) and heme oxygenase-1(HO-1) were also studied. BRE has an inhibitory effect on lipid accumulation and caused acute hepatotoxicity manifested by increased levels of lipid peroxidation, serum alanine aminotransferase, and aspartate aminotransferase with corresponding histopathological changes and high levels of oxidative stress. BRE treatment significantly increased the level of CYP2E1, Nrf-2, and HO-1 in a dose-dependent manner. Besides, 3-(E)-(methylthio)methylene-2-pyrrolidinethione significantly increased radical-scavenging effects and the expression of Nrf-2 in oleic acid-treated HepG2 cells. These results suggest that BRE treatment reduces lipid accumulation in oleic acid-induced steatosis of HepG2 cells, and has a hepatoprotective effect against CCl4-induced liver injury in rats, possibly through Nrf-2/HO-1-mediated antioxidant effects.

Allyl isothiocyanate reduces liver fibrosis by regulating Kupffer cell activation in rats.

Allyl isothiocyanate (AITC), a metabolite of the glucosinolate sinigrin, protects the liver of rats injured by carbon tetrachloride (CCl4). This study evaluated whether AITC reduces hepatic fibrosis in rats repetitively exposed to CCl4. Serum chemistry showed that AITC (doses of 5 and 50 mg) administered to rats exposed to CCl4 significantly reduced the levels of alanine aminotransferase and aspartate aminotransferase activity that were elevated in CCl4-intoxicated rats. The connective tissue in AITC-treated rats was significantly reduced based on Sirius staining. In addition, Kupffer cell activation was significantly reduced in the AITC and CCl4 co-treated groups. Collectively, this study suggests that AITC mitigates hepatic fibrosis in rats repetitively exposed to CCl4 with concurrent regulation of Kupffer cell and monocyte activation.

Beetroot (Beta vulgaris) rescues mice from γ-ray irradiation by accelerating hematopoiesis and curtailing immunosuppression.

CONTEXT: Beetroot [Beta vulgaris Linné (Chenopodiaceae)], a vegetable usually consumed as a food or a medicinal plant in Europe, has been reported to have antioxidant and anti-inflammatory properties. Since the lymphohematopoietic system is the most sensitive tissue to ionizing radiation, protecting it from radiation damage is one of the best ways to decrease detrimental effects from radiation exposure. OBJECTIVE: In this study, we evaluated the radio-protective effects of beetroot in hematopoietic stem cells (HSCs) and progenitor cells. MATERIALS AND METHODS: Beetroot extract was administered at a dose of 400 mg/mouse per os (p.o.) three times into C57BL/6 mice and, at day 10 after γ-ray irradiation, diverse molecular presentations were measured and compared against non-irradiated and irradiated mice with PBS treatments. Survival of beetroot-fed and unfed irradiated animal was also compared. RESULTS: Beetroot not only stimulated cell proliferation, but also minimized DNA damage of splenocytes. Beetroot also repopulated S-phase cells and increased Ki-67 or c-Kit positive cells in bone marrow. Moreover, beetroot-treated mice showed notable boosting of differentiation of HSCs into burst-forming units-erythroid along with increased production of IL-3. Also, beetroot-treated mice displayed enhancement in the level of hematocrit and hemoglobin as well as the number of red blood cell in peripheral blood. Beetroot diet improved survival rate of lethally exposed mice with a dose reduction factor (DRF) of 1.1. DISCUSSION AND CONCLUSION: These results suggest that beetroot has the potency to preserve bone marrow integrity and stimulate the differentiation of HSCs against ionizing radiation.

Anti-inflammatory activities of Dangyuja (Citrus grandis Osbeck) in concanavalin A stimulated murine splenocytes and 12-O-tetradecanoylphorbol-13-acetate-induced murine skin edema.

Dangyuja (Citrus grandis Osbeck), a citrus cultivated in southern Korea, has been used in traditional medicine for its anti-inflammatory effect. In this study, we investigated the anti-inflammatory potential of extract of Citrus grandis Osbeck (ECGO). In in vitro assays, ECGO treatment of concanavalin A (10µg/ml, for 24h) stimulated splenocytes showed significant reduction in CD44/CD62L+ T cell population and a marked decrease in the production of inflammatory cytokines IL-2, IFN-γ and IL-4. Interestingly, in vivo assays of ECGO topical treatment (100µg/20µl/ear) significantly mitigated the TPA (4µg/20µl/ear) induced edema induction and Myeloperoxidase activity. Anti-inflammatory potential of ECGO were further evidenced through its potent decrease in expression of inducible nitric oxide, cyclooxygenase-2, IL-1ß and TNF-α and suppressed homing of CD3+ T cells and F4/80+ macrophages to site of inflammation. This study emphasizes the possibility of developing ECGO as an alternative natural topical agent to combat inflammatory diseases.

Hepatoprotective effects of allyl isothiocyanate against carbon tetrachloride-induced hepatotoxicity in rat.

We evaluated the hepatoprotective activity of allyl isothiocyanate (AITC) against carbon tetrachloride (CCl4)-induced liver injury in rats. Sprague Dawley rats were orally administered AITC at doses of 5 (AITC 5) and 50 (AITC 50) mg/kg body weight once daily for 3 days, with or without intraperitoneal injection of CCl4. Serum chemistry was assessed for changes in alanine aminotransferase (ALT) and aspartate aminotransferase (AST). The enzyme activities of superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA) were examined in liver tissues, while pro-inflammatory cytokines including tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1ß) mRNA expression were analyzed using real-time polymerase chain reaction. And heme oxygenase-1 (HO-1) and ionized calcium binding protein-1 (Iba-1) immunoreactivities were evaluated by Western blot analysis and immunohistochemistry, respectively. In serum chemistry, the oral administration of AITC itself did not affect the serum levels of ALT or AST, furthermore pretreatment with AITC 5 and AITC 50 significantly reduced the ALT and AST activity levels that were elevated in CCl4-intoxicated rats. In addition, AITC significantly suppressed the reduction of SOD and CAT, and the elevation of MDA, TNF-α mRNA expression, on the other hands, induced the expression of HO-1 compared with those of the vehicle-treated CCl4 group. The histopathological evaluation and Iba-1 immunoreactivity also supported the hepatoprotective effects of AITC against CCl4-induced liver injury. These results suggest that AITC ameliorates oxidative liver injury, possibly through reducing lipid peroxidation, enhancing antioxidant enzymes, and suppressing Kupffer cells and macrophages.

Citrus hallabong [(Citrus unshiu × C. sinensis) × C. reticulata)] exerts potent anti-inflammatory properties in murine splenocytes and TPA-induced murine ear oedema model.

CONTEXT: Hallabong [(Citrus unshiu × C. sinensis) X C. reticulata)] (Rutaceae) is a hybrid citrus cultivated in temperate regions of South Korea. Its fruit is well-known for pharmacological properties. OBJECTIVE: This study examined the anti-inflammatory effect of 80% ethanol extract of Hallabong (HE) on concanavalin A (Con A)-stimulated splenocytes and mouse oedema model induced by 12-O-tetradecanoylphorbal acetate (TPA). MATERIALS AND METHODS: Murine splenocytes treated with HE were stimulated with Con A (10 µg/mL, for 24 h) were evaluated for T-cell population and production of inflammatory cytokines IL-2, IL-4 and IFN-γ. Anti-inflammatory effect of topically applied HE (100 µg/20 µL) on TPA (4 µg/20 µL/ear)-induced ear oedema was investigated in mouse model. RESULTS: HE-treated Con A-stimulated murine splenocytes showed a marked decrease in CD44/CD62L+ memory T-cell population, an important marker for anti-inflammatory activity, and a significant inhibition in the production of IL-2 and IFN-γ. HE treatment had reduced the mouse skin oedema (47%) and myeloperoxidase (MPO) activity significantly (40%) in TPA-challenged tissues. More importantly, immunohistochemical localization revealed the suppressed (p < 0.05) expression of inducible nitric oxide (iNOS), cyclooxygenase-2 (COX2). HE decreased the infiltration of CD3+ T cells and F4/80+ macrophages to the site of inflammation and a topical application of HE significantly suppressed the expression of TNF-α (20.2%). DISCUSSION AND CONCLUSION: A topical application of HE can exert a potential anti-inflammatory effect and HE can be explored further as a putative alternative therapeutic agent for inflammatory oedema.

Diphlorethohydroxycarmalol attenuated cell damage against UVB radiation via enhancing antioxidant effects and absorbing UVB ray in human HaCaT keratinocytes.

Exposure of human skin to excessive ultraviolet B (UVB) radiation induces pathophysiological processes via the generation of reactive oxygen species (ROS) in skin cells, such as keratinocytes. This study investigated the ability of diphlorethohydroxycarmalol (DPHC) to protect human keratinocytes (HaCaT) against UVB-induced cell damage. DPHC restored cell viability that was reduced by UVB light. DPHC had an absorption maximum close to the UVB spectrum and decreased UVB-induced intracellular ROS levels, increased levels of reduced glutathione, activated superoxide dismutase and catalase. DPHC also decreased UVB-mediated damage to cellular components, including lipids, proteins, DNA, and attenuated UVB-induced apoptosis. These results suggest that DPHC safeguards human keratinocytes against UVB-induced cell damage by absorbing UVB ray, scavenging ROS and enhancing antioxidant system.

Anti-melanogenesis constituents from the seaweed Dictyota coriacea.

This study was conducted to identify the anti-melanogenesis constituents from a seaweed Dictyota coriacea (Holmes). Three known compounds, viz. 1,9-dihydroxycrenulide (1), epiloliolide (2) and D-mannitol (3), were isolated from the ethanol extract. The melanin synthesis inhibition activities were evaluated using B16F10 melanoma cells for the isolates. Compared with the positive control, arbutin, compounds 1 and 2 exhibited more potency, showing 27.8 and 22.6% inhibition activities at a substrate concentration of 30 microg/mL. Our studies also indicate that these compounds are not cytotoxic. Hence, they might prove to be useful therapeutic agents for treating hyperpigmentation and effective components of whitening cosmetics.

Neuroprotective effect of methyl lucidone against microglia-mediated neurotoxicity.

Excessive microglial activation-mediated neurotoxicity has been implicated in playing a crucial role in the pathogenesis of stroke and neurodegenerative diseases. Therefore, much attention has been paid to therapeutic strategies aimed at suppressing neurotoxic microglial activation. The microglial regulatory mechanism of methyl lucidone, a cyclopentenedione isolated from the stem bark of Lindera erythrocarpa Makino, was investigated in the present study. Methyl lucidone treatment (0.1-10 µM) significantly inhibited lipopolysaccharide (LPS, 100 ng/ml, 24 h)-stimulated nitric oxide (NO) production in a dose-dependent manner in both primary cortical microglia and BV-2 cell line. Moreover, it strongly inhibited LPS-stimulated secretion of pro-inflammatory cytokines, such as interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α). Methyl lucidone treatment markedly induced down-regulation of LPS-induced nuclear translocation of nuclear factor κB (NF-κB) through preventing the degradation of the inhibitory protein IκBα. In addition, phosphorylation of Akt and mitogen-activated protein kinases (MAPKs) such as extracellular signal-regulated kinase (ERK) and p38 kinases were also suppressed by methyl lucidone. The cell viabilities of HT-22 neurons were significantly attenuated by treatment of the conditioned media containing neurotoxic secretary molecules from LPS-stimulated microglia. However, methyl lucidone significantly blocked neuronal cell death induced by microglial conditioned media. These neuroprotective effects of methyl lucidone were also confirmed in a neuron-microglia co-culture system using EGFP-transfected B35 neuroblastoma cell line. Taken together, these results suggest that methyl lucidone may have a neuroprotective potential via inhibition of neurotoxic microglial activation implicated in neurodegeneration.

Sasa quelpaertensis phenylpropanoid derivative suppresses lipopolysaccharide-induced nitric oxide synthase and cyclo-oxygenase-2 expressions in RAW 264.7 cells.

3-O-p-Coumaroyl-1-(4-hydroxy-3,5-dimethoxyphenyl)-1-O-ß-D-gulcopyranosylpropanol (ESQ10) is a naturally occurring phenylpropanoid derivative isolated from Sasa quelpaertensis (Gramineae). In the present study, we discovered that ESQ10 inhibits nitric oxide (NO) and prostaglandin E(2) (PGE(2)) production in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. ESQ10 attenuated LPS-induced synthesis of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in parallel and inhibited LPS-induced interleukin-6 production, as determined by an enzyme-linked immunosorbent assay in the macrophages. The mechanism of the antiinflammatory action of ESQ10, i.e., suppression of nuclear factor (NF)-κB and mitogen-activated protein kinase activation, has been documented. However, ESQ10 could not influence LPS-mediated IκB-α degradation and extracellular signal-regulated kinase/c-Jun amino-terminal kinase phosphorylation at concentrations of up to 373 µM. To test the potential application of ESQ10 as a topical material, we also conducted a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay on human HaCaT keratinocytes as well as human dermal fibroblast cells. In this assay, ESQ10 did not induce cytotoxicity. Taken together, the results suggest that ESQ10 may be considered an antiinflammatory candidate for treating inflammatory and skin diseases.

Compounds with tyrosinase inhibition, elastase inhibition and DPPH radical scavenging activities from the branches of Distylium racemosum Sieb. et Zucc.

Twenty compounds were isolated from the ethanol extract of Distylium racemosum branches and their inhibitory activities on tyrosinase, elastase and free radicals evaluated. The isolated compounds were identified as dibenzofurans (1-4), abscisic acid (5), 6'-O-galloylsalidroside (6), catechin derivatives (7-11), gallic acid derivatives (12-14), tyrosol (15), flavonoids (16-18), lupeol (19) and 1,2,3,6-tetragalloylglucose (20). For study of tyrosinase inhibition activities, when compared with arbutin (IC(50) 48.8 µg/mL), four compounds (8, 11, 13, 17) showed higher activities, with IC(50) values of 4.8, 30.2, 40.5 and 37.7 µg/mL, respectively. For the elastase inhibition test, dibenzofuran 1 showed greater activity than the positive control, oleanolic acid (IC(50) 9.7 µg/mL), with an IC(50) of 7.7 µg/mL. In the studies on DPPH radical scavenging activities, five compounds (11, 12, 13, 14, 15) showed higher activities than ascorbic acid (IC(50) 5.0 µg/mL), with IC(50) values of 4.6, 3.9, 2.9, 3.8 and 4.7 µg/mL, respectively.

Diphlorethohydroxycarmalol, isolated from the brown algae Ishige okamurae, protects against radiation-induced cell damage in mice.

The aim of this study was to evaluate the radioprotective effects of diphlorethohydroxycarmalol (DPHC), isolated from the brown algae Ishige okamurae, in mice subjected to gamma irradiation. DPHC significantly decreased the level of radiation-induced intracellular reactive oxygen species in cultured Chinese hamster lung fibroblast (V79-4) cells (p < 0.05), enhanced cell viability that decreased after exposure to γ-rays, and reduced radiation-induced apoptosis in the V79-4 cells. Pretreatment with DPHC (100 mg/kg) in mice prior to irradiation significantly protected the intestinal crypt cells in the jejunum (p < 0.01) and maintained villi height (p < 0.01), compared with those of the vehicle-treated irradiated group. Mice pretreated with DPHC also exhibited dose-dependent increases in the bone marrow cell viability. The dose-reduction factor for gamma irradiation in the DPHC-pretreated mice was 2.05 at 3.5 days after irradiation. These results suggest that DHPC plays a role in protecting cells from irradiation-induced apoptosis, through the scavenging of reactive oxygen species in vitro, and that DPHC significantly protected intestinal progenitor cells and bone marrows cells that were decreased by gamma irradiation in vivo.

Neolitsea sericea essential oil attenuates LPS-induced inflammation in RAW 264.7 macrophages by suppressing NF-kappaB and MAPK activation.

The chemical composition and anti-inflammatory 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-beta-ocimene (13.3%), beta-caryophyllene (4.8%), and 4-terpineol (4.2%). The effects of NSE on nitric oxide (NO), prostaglandin E2 (PGE2), tumour necrosis factor (TNF)-alpha, interleukin (IL)-1beta, 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-kappaB (NF-kappaB) activation and the phosphorylation of mitogen-activated protein kinases (MAPK). NSE inhibited NF-kappaB activation by LPS, and this was associated with the abrogation of IkappaB-alpha 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 anti-inflammatory effects in LPS-stimulated RAW 264.7 macrophages by inhibition of NF-kappaB activation and MAPK phosphorylation, and, therefore, may be useful for treatment of inflammatory diseases.

Effect of palmitoleic acid on melanogenic protein expression in murine b16 melanoma.

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.

Inhibition of nitric oxide production in lipopolysaccharide-activated RAW 264.7 macrophages by Jeju plant extracts.

Nitric oxide (NO) produced in large amounts by inducible nitric oxide synthase (iNOS) is known to be responsible for the vasodilation and hypotension observed during septic shock and inflammation. Thus, inhibitors of iNOS may be useful candidates for the treatment of inflammatory diseases accompanied by the overproduction of NO. In this study, we prepared alcoholic extracts of Jeju plants and screened them for their inhibitory activity against NO production in lipopolysaccharide (LPS)-activated macrophages. Among the 260 kinds of plant extract tested, 122 extracts showed potent inhibitory activity towards NO production by more than 25% at a concentration of 100 µg/mL. Plants such as Malus sieboldii, Vaccinium oldhamii, Corylus hallaisanensis, Carpinus laxiflora, Styrax obassia, and Securinega suffruticosa showed the most potent inhibition (above 70%) at a concentration of 100 µg/mL. The cytotoxic effects of the plant extracts were determined by colorimetric MTT assays and most plant extracts exhibited only moderate cytotoxicity at 100 µg/mL. Therefore, these plants should be considered promising candidates for the further purification of bioactive compounds and would be useful for the treatment of inflammatory diseases accompanying overproduction of NO.