Ethyl acetate fraction of Osmanthus fragrans var. aurantiacus and its triterpenoids suppress proliferation and survival of colorectal cancer cells by inhibiting NF-κB and COX2.

ETHNOPHARMACOLOGICAL RELEVANCE: Colorectal cancer (CRC) remains a significant global health concern, and targeting inflammation has emerged as a promising approach for its prevention and treatment. Medicinal plants and phytochemicals have garnered attention for their potential efficacy against inflammation with minimal toxicity. Osmanthus fragrans var. aurantiacus Makino (O. fragrans) has a history of traditional use in Korea and China in treating various inflammation-related conditions, but its potential use for CRC has not been uncovered. AIM OF THE STUDY: This study aims to explore the potential anti-proliferative and pro-apoptotic properties of O. fragrans, focusing on its impact on CRC treatment. By investigating O. fragrans, we aim to uncover its anti-proliferative and apoptotic effects in human CRC cells, potentially paving the way for effective and well-tolerated therapeutic strategies for CRC patients. MATERIALS AND METHODS: Ethanol (EtOH) extracts of O. fragrans leaf and flower, along with specific fractions (n-hexane, ethyl acetate (EtOAc), n-butanol, and the aqueous residue) were evaluated for their anti-proliferative effects in human CRC cells using MTT assays, and compared to normal colon cells. Mechanistic insights and chemical profiling were obtained through flow cytometry, colorimetric assays, western blotting, and molecular docking, and high-performance liquid chromatography (HPLC) system. RESULTS: Both flower and leaf EtOH extracts of O. fragrans exhibited significant anti-proliferative effects in human CRC cells, with the leaf extract demonstrating higher potency. The EtOAc fraction from the leaf extract displayed the strongest anti-CRC cell proliferative effects while no cytotoxic effects in normal colon cells. Chemical profiling of these fractions identified triterpenoids as significant components in the EtOAc fractions. The leaf EtOAc fraction caused cell cycle arrest and apoptosis, accompanied by elevating intracellular reactive oxygen species and mitochondrial dysfunction in CRC cells. Additionally, it inhibited NF-κB and ERK1/2 signaling, leading to reduced COX2 expression. Notably, two triterpenoids isolated from the leaf EtOAc fraction, maslinic acid and corosolic acid, displayed potent anti-cancer activity in CRC cells without affecting normal colon cells. Corosolic acid exhibited a strong binding affinity to COX2 and reduced its expression, supporting its role in the anti-inflammatory and anti-cancer effects. CONCLUSIONS: Our findings suggest that O. fragrans, particularly its triterpenoid-rich EtOAc fraction, holds promise as a novel therapeutic agent for CRC prevention and therapy. These results provide valuable insights into the potential application of O. fragrans and its bioactive compounds in combating CRC.

Bioactivity-Guided Fraction from Viscera of Abalone, Haliotis discus hannai Suppresses Cellular Basophils Activation and Anaphylaxis in Mice.

Basophils and mast cells are specialized effector cells in allergic reactions. Haliotis discus hannai (abalone), is valuable seafood. Abalone male viscera, which has a brownish color and has not been previously reported to show anti-allergic activities, was extracted with acetone. Six different acetone/hexane fractions (0, 10, 20, 30, 40, and 100%) were obtained using a silica column via ß-hexosaminidase release inhibitory activity-guided selection in phorbol myristate acetate and a calcium ionophore, A23187 (PMACI)-induced human basophils, KU812F cells. The 40% acetone/hexane fraction (A40) exhibited the strongest inhibition of PMACI-induced-ß-hexosaminidase release. This fraction dose-dependently inhibited reactive oxygen species (ROS) production and calcium mobilization without cytotoxicity. Western blot analysis revealed that A40 down-regulated PMACI-induced MAPK (ERK 1/2, p-38, and JNK) phosphorylation, and the NF-κB translocation from the cytosol to membrane. Moreover, A40 inhibited PMACI-induced interleukin (IL)-1ß, IL-6, and IL-8 production. Anti-allergic activities of A40 were confirmed based on inhibitory effects on IL-4 and tumor necrosis factor alpha (TNF-α) production in compound (com) 48/80-induced rat basophilic leukemia (RBL)-2H3 cells. A40 inhibited ß-hexosaminidase release and cytokine production such as IL-4 and TNF-α produced by com 48/80-stimulated RBL-2H3 cells. Furthermore, it's fraction attenuated the IgE/DNP-induced passive cutaneous anaphylaxis (PCA) reaction in the ears of BALB/c mice. Our results suggest that abalone contains the active fraction, A40 is a potent therapeutic and functional material to treat allergic diseases.

Anti-Inflammatory Food in Asthma Prepared from Combination of Raphanus sativus L., Allium hookeri, Acanthopanax sessiliflorum, and Dendropanax morbiferus Extracts via Bioassay-Guided Selection.

Asthma is a highly prevalent inflammatory disease of the respiratory airways and an increasing health risk worldwide. Hence, finding new strategies to control or attenuate this condition is necessary. This study suggests nutraceuticals that are a combination of herbal plant extracts prepared from Acanthopanax sessiliflorum (AS), Codonopsis lanceolate (CL), Dendropanax morbiferus (DM), Allium hookeri (AH), and Raphanus sativus L. (RS) that can improve immunomodulatory ability through the detoxification and diuresis of air pollutants. Herbal parts (AH whole plant, RS and CL roots, AS and DM stems, and DM leaves) were selected, and four types of mixtures using plant extracts were prepared. Among these mixtures, M2 and M4 exhibited antioxidant activities in potent 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) and 1,1-diphenyl-ß-picrylhydrazine (DPPH) radical assays. Moreover, M4 exhibited a marked increase in glutathione S-transferase (GST) activity and significantly inhibited the inflammatory mediator, nitric oxide (NO) and proinflammatory cytokines, interleukin (IL)-1ß, IL-6 and tumor necrosis factor (TNF)-α generation. Furthermore, M4 exhibited the strongest antioxidant, hepatoprotective, and anti-inflammatory effects and was selected to prepare the product. Before manufacturing the product, we determined that the active mixture, M4, inhibited gene expression and generation of proinflammatory cytokines IL-1ß, IL-6 and TNF-α in ovalbumin (OVA)-, lipopolysaccharide (LPS)-, and particulate matter (PM)-induced asthmatic rat models. The granular product (GP) was manufactured using M4 along with additives, i.e., lactose, oligosaccharide, stevioside extract, and nutmeg seed essential oils (flavor masking), in a ratio of 1:4 using a granulation machine, dried and ultimately packaged. The GP inhibited the generation of proinflammatory cytokines IL-1ß, IL-6 and TNF-α in OVA-, LPS- and PM-induced asthmatic rat models. These results suggest that GP prepared from a combination of herbal plants (AS, CL, DM, AH and RS) is a potent functional food with anti-inflammatory activity that can be used to treat asthma caused by ambient air pollutants.

Sargahydroquinoic acid isolated from Sargassum serratifolium as inhibitor of cellular basophils activation and passive cutaneous anaphylaxis in mice.

Basophils and mast cells are characteristic effector cells in allergic reactions. Sargahydorquinoic acid (SHQA), a compound isolated from Sargassum serratifolium (marine alga), possesses various biochemical properties, including potent antioxidant activities. The objective of the present study was to investigate inhibitory effects of SHQA on the activation of human basophilic KU812F cells induced by phorbol myristate acetate and A23187 (PMACI), a calcium ionophore. Furthermore, we confirmed the inhibitory effects of SHQA on the activation of rat basophilic leukemia (RBL)-2H3 cells induced by compound 48/80 (com 48/80), bone marrow-derived mast cells (BMCMCs) induced by anti-dinitrophenyl(DNP)-immunoglobulin E (IgE)/DNP-bovine serum albumin (BSA), DNP/IgE and on the reaction of passive cutaneous anaphylaxis (PCA) mediated by IgE. SHQA reduced PMACI-induced intracellular reactive oxygen species (ROS) and calcium levels. Western blot analysis revealed that SHQA downregulated the activation of ERK, p38, and NF-κB in a dose-dependent manner. Moreover, SHQA suppressed the production and gene expression of various cytokines, including interleukin (IL)-1 ß, IL-4, IL-6, and IL-8 in PMACI-induced KU812F cells and IL-4 and tumor necrosis factor (TNF)- α in com 48/80-induced RBL-2H3 cells. It also determined the inhibition of PMACI, com 48/80- and IgE/DNP-induced degranulation by reducing the release of ß -hexosaminidase. Furthermore, it attenuated the IgE/DNP-induced PCA reaction in the ears of BALB/c mice. These results suggest that SHQA isolated from S. serratifolium is a potential therapeutic functional food material for inhibiting effector cell activation in allergic reactions and anaphylaxis in animal model.

Antioxidant Compounds, Kirenol and Methyl ent-16α, 17-dihydroxy-kauran-19-oate Bioactivity-Guided Isolated from Siegesbeckia glabrescens Attenuates MITF-Mediated Melanogenesis via Inhibition of Intracellular ROS Production.

Siegesbeckia glabrescens (Compositae), an annual herb indigenous to Korean mountainous regions and has been eaten as a food in Korea. This study investigated ABTS, DPPH and nitric oxide (NO) radical-scavenging activities, and melanin production and TYR inhibitory effects-guided fractionation to identify therapeutic phytochemicals from S. glabrescens that can attenuate oxidation and melanogenesis in murine melanoma B16F10 cells. Nine compounds with inhibitory effects on melanin production, and TYR activity, and ABTS, DPPH, and NO radical scavenging activity were isolated from the 100% ethanol fraction from S. glabrescens. Among the nine compounds, kirenol (K), methyl ent-16α, 17-dihydroxy-kauran-19-oate (MDK) had strong inhibitory effects on melanin production and TYR activity with antioxidant effects. Western blot analysis revealed that K and MDK suppressed tyrosinase-related protein (TYRP)-1, TYRP-2 and microphthalmia-associated transcription factor (MITF) expression. Moreover, these two compounds inhibited intracellular reactive oxygen species (ROS) level in tert-butyl hydroperoxide (t-BHP)-treated B16F10 cells. Our results suggest that S. glabrescens containing active compounds such as K and MDK, which has antioxidant and antimelanogenesis effects, is the potent therapeutic and functional material for the prevention of oxidation-induced hyperpigmentation.

Anti-inflammatory activity of phenylpropyl triterpenoids from Osmanthus fragrans var. aurantiacus leaves.

Osmanthus fragrans var. aurantiacus has been used for the treatment of menopausal pain, foul breath, and intestinal bleeding. Four phenylpropyl triterpenoids, 3-O-trans-p-coumaroyltormentic acid (1), 3ß-trans-p-coumaroyloxy-2α-hydroxyl-urs-12-en-28-oic acid (2), 3ß-cis-p-coumaroyloxy-2α-hydroxyl-urs-12-en-28-oic acid (3), 3-O-cis-coumaroylmaslinic acid (4), were isolated from the leaves of O. fragrans var. aurantiacus and the inhibitory effect on nitric oxide (NO) production in lipopolysaccharide (LPS)-induced macrophages were evaluated. Among them, compounds 2-4 concentration dependently showed NO production inhibitory effects. To determine the signaling factors involved in the inhibition of NO production by compounds 2-4, we assessed anti-inflammatory activity. Western blot analysis revealed compounds 2-4 significantly decreased the expression of LPS-stimulated protein, inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2, nuclear factor-kappa B (NF-κB) and phosphorylated extracellular regulated kinase (pERK)1/2. Also, compounds 2-4 downregulated tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, IL-6 and IL-8 levels in LPS-induced macrophages and colonic epithelial cells. This study demonstrated that phenylpropyl triterpenoids 2-4 isolated from O. fragrans var. aurantiacus leaves can be used as potential candidates for the prevention and treatment of inflammatory bowel disease.

Eckol from Ecklonia cava Suppresses Immunoglobulin E-mediated Mast Cell Activation and Passive Cutaneous Anaphylaxis in Mice.

Eckol, a precursor compound belonging to the dibenzo-1,4-dioxin class of phlorotannins, is a phloroglucinol derivative that exerts various activities. In the present study, we investigated the antiallergic effects of eckol isolated from the marine brown algae, Ecklonia cava using immunoglobulin E (IgE)/bovine serum albumin (BSA)-stimulated mouse bone marrow-derived cultured mast cells (BMCMC) and a mouse model of anaphylaxis. Eckol inhibited IgE/BSA-induced BMCMC degranulation by reducing ß-hexosaminidase release. A flow cytometric analysis revealed that eckol decreases FcεRI expression on cell surface and IgE binding to the FcεRI in BMCMC. Moreover, eckol suppressed the production of the cytokines, interleukin (IL)-4, IL-5, IL-6, and IL-13 and the chemokine, thymus activation-regulated chemokine (TARC) by downregulating, IκB-α degradation and NF-κB nuclear translocation. Furthermore, it attenuated the passive cutaneous anaphylactic reaction induced by IgE/BSA-stimulation in the ear of BALB/c mice. These results suggest that eckol is a potential therapeutic candidate for the prevention and treatment of allergic disorders.

p-Hydroxybenzoic Acid ß-d-Glucosyl Ester and Cimidahurinine with Antimelanogenesis and Antioxidant Effects from Pyracantha angustifolia via Bioactivity-Guided Fractionation.

This study evaluated bioactivity-guided fractionation as a means to identify therapeutic phytochemicals from Pyracantha angustifolia that can attenuate melanogenesis and oxidation. Seven compounds with inhibitory effects on melanin production and tyrosinase (TYR) activity, and ABTS and DPPH radical-scavenging activities, which have not been reported as whitening materials, were isolated from the n-butanol fraction from P. angustifolia leaves (PAL). Among the seven compounds, p-hydroxybenzoic acid ß-d-glucosylester (HG), and cimidahurinine (CH) had strong inhibitory effects on melanin production and TYR activity, as well as ABTS and DPPH radical-scavenging activities. Western blot analysis showed that HG and CH suppressed tyrosinase-related protein (TYRP)-1 and TYRP-2 expression. Moreover, HG and CH inhibited reactive oxygen species (ROS) generation in tert-butyl hydroperoxide (t-BHP)-treated B16F10 cells. These results suggest that P. angustifolia containing active compounds, such as HG and CH, is a potent therapeutic candidate for the development of hypopigmenting agents.

Anti-inflammatory and anti-oxidative activities of lemon myrtle (Backhousia citriodora) leaf extract.

In this present study, we examined the anti-inflammatory and anti-oxidative properties of alcoholic lemon myrtle extract (LME). The total polyphenol and flavonoid content of LME were determined as 118.77 and 14.53 mg/g extract, respectively. LME showed anti-oxidative properties, such as 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) and 2,2'-azino-bis(3-ethylbenz-thiazoline-6-sulfonic acid) (ABTS) radical scavenging activity. The anti-inflammatory activities of LME were investigated using the lipopolysaccharide-stimulated murine macrophage RAW 264.7 cells. Pretreatment with LME was performed at non-cytotoxic concentrations of 10-100 µg/mL. LME inhibited the production of inflammatory mediators such as nitric oxide (NO). Enzyme-linked immunosorbent assay and reverse-transcriptase polymerase chain reaction (RT-PCR) revealed that pretreatment with LME suppressed the protein expression and mRNA levels of pro-inflammatory cytokines such as interleukin IL-6, and tumor necrosis factor (TNF)-α in a concentration-dependent manner, respectively. These results suggest that LME could be used as a potential therapeutic agent having potent anti-inflammatory effects that could be used to treat inflammatory bowel disease.

Inhibitory Effects of Eriodictyol-7-O-ß-d-glucuronide and 5,7-Dihydroxy-4-chromene Isolated from Chrysanthemum zawadskii var. latilobum in FcεRI-Mediated Human Basophilic KU812F Cell Activation.

Chrysanthemum zawadskii var. latilobum (CZL) has been used in Eastern medicine for the treatment of various diseases, such as pneumonia, bronchitis, cough, the common cold, pharyngitis, bladder-related disorders, gastroenteric disorders, and hypertension. In the present study, we isolated two strong antiallergic compounds from CZL, namely, eriodictyol-7-O-ß-d-glucuronide (EDG) and 5,7-dihydroxy-4-chromene (DC), and investigated their antiallergic effects in FcεRI-mediated human basophilic KU812F cells. EDG and DC downregulated the protein and messenger RNA (mRNA) expression of FcεRI on the cell surface. Moreover, Western blotting analysis showed that EDG and DC inhibited the expression of protein tyrosine kinases such as Syk and Lyn, and extracellular-regulated kinases (ERK) 1/2. These results suggested that EDG and DC, antiallergic constituents of CZL, are potential therapeutic candidates for protection against and for the treatment of allergic disorders.

Pandanus fascicularis Lam Extract Inhibits Pro-Inflammatory Cytokines Production in LPS-Stimulated RAW 264.7 Cells.

Pandanus fascicularis Lam has been shown to exert to a variety of physiological effects on edema, tumors, leprosy, spasm, inflammation, pain, and rheumatoid arthritis. In this study, the effects of a P. fascicularis extract on the production of lipopolysaccharide (LPS)-induced pro-inflammatory cytokines interleukin (IL)-1ß, IL-6, and tumor necrosis factor (TNF)-α expression and on the production of the inflammatory mediator, nitric oxide (NO), in RAW 264.7 cells were examined. The P. fascicularis extract decreased LPS-induced NO production. Moreover, enzyme-linked immunosorbent assays revealed that the P. fascicularis extract concentration-dependently suppressed LPS-induced production of the pro-inflammatory cytokines IL-1ß, IL-6, and TNF-α. These results suggest that the P. fascicularis extract has potential as an anti-inflammatory therapeutic substance for use in the prevention of the inflammatory disorder.

Suppressive Effects of Vaccinium angustifolium Root Extract via Down-Regulation of Activation of Syk, Lyn, and NF-κB in FcɛRI-Mediated Allergic Reactions.

Vaccinium angustifolium, reported as the lowbush blueberry, has a rich polyphenolic content with which biological activities have been closely associated. In this study, the effects of V. angustifolium root extract (VAE) on the anti-FcɛRI α chain antibody (CRA-1)-induced FcɛRI-mediated signaling factors, protein tyrosine kinases (PTK), Lyn, Syk, and nuclear factor kappa-B cells (NF-κB) in KU812F cells were investigated. The total phenolic content of VAE was found to be 170±1.9 mg gallic acid equivalents/g. Western blot analysis revealed that VAE dose-dependently inhibited FcɛRI-mediated phosphorylation of PTK involving Lyn and Syk. Evaluation of intracellular reactive oxygen species (ROS) by spectrofluorometric analysis using 2'7'-dichlorofluorescin-diacetate revealed that they were reduced by VAE in a dose-dependent manner. Moreover, VAE reduced the levels of ß-hexosaminidase released from CRA-1-stimulated KU812F cells. It was identified that VAE suppressed CRA-1-induced activation of NF-κB by Western blot analysis. Our results show that VAE may contribute to the inhibition of allergic actions via inactivation of basophils through the inhibition of ß-hexosaminidase release and ROS production, which occurs as a result of inhibition of PTK, Syk, Lyn, and NF-κB.

Biflavonoids Isolated from Selaginella tamariscina and Their Anti-Inflammatory Activities via ERK 1/2 Signaling.

Selaginella tamariscina (S. tamariscina) (Beauv.) Spring (Selaginellaceae) has been used in oriental medicine for the treatment of dysmenorrhea, chronic hepatitis, hyperglycemia, amenorrhea, hematuria, prolapse of the anus and metrorrhagia. In the present study, we isolated two strong anti-inflammatory compounds, the biflavonoids hinokiflavone (H) and 7'-O-methyl hinokiflavone (mH), from S. tamariscina and examined their anti-inflammatory activities in lipopolysaccharide (LPS)-mediated murine macrophages (RAW 264.7) and colon epithelial cells (HT-29). H and mH suppressed the production of the inflammatory mediators nitric oxide (NO), interleukin (IL)-6, IL-8, and tumor-necrosis factor (TNF)-α, which are most highly activated in inflammatory bowel disease (IBD). In addition, Western blot analysis revealed that H and mH suppressed the LPS-induced expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2, and the activation of nuclear factor-κB (NF-κB) and extracellular regulated kinases (ERK) 1/2. These results suggest that H and mH are compounds having potent anti-inflammatory effects that could be used to treat such diseases as IBD.

Reduction in Prevalence of Hypertension and Blood Heavy Metals among Curry-Consumed Korean.

Curcuma longa is rich in curcumin and used to treat disease and in food as a spice, especially in curry dishes. The objective of the present study was to determine whether curry intake reduces blood levels of heavy metals and hypertension (HTN) using Korean National Health and Nutrition Examination Survey 2013 data (n = 1,350). Study subjects (n = 1,350) were divided into two groups: 1) a curry intake group (n = 603), members of which had consumed a curry dish more than once a month over the previous year, and 2) a non-curry intake group (n = 747). Analysis showed the curry intake group had significantly lower heavy metal blood concentrations (Pb, Hg, and Cd) and blood concentrations of heavy metals were significantly associated with prevalence of HTN (P < 0.001 for Pb, Hg, and Cd). Curry intake also reduced the risk of HTN prevalence (odd ratios for curry intake vs. non-curry intake; Pb, 1.503 vs. 1.862; Hg, 1.112 vs. 1.149; Cd, 1.676 vs. 1.988). Logistic regression analysis was used to confirm the association between blood heavy metal levels and HTN. After adjusting for age, sex, lifetime smoking, and body mass index, the odd ratio of HTN was significant in the non-curry intake group, but not in the curry intake group, implying other factors influenced relations in the curry intake group. In view of the importance of curry consumption with reduced concentrations of heavy metals in blood and the prevalence of HTN, we suggest further well-designed clinical trials be conducted.

Anti-inflammatory activity of mulberrofuran K isolated from the bark of Morus bombycis.

Morus bombycis Koidzumi, commonly known as silkworm mulberry, is a plant belonging to family Moraceae. It has been used in Asian countries as a traditional medicine for treating hypertension, diabetes, and inflammatory disorders. In this study, we isolated eleven compounds from the cortex of M. bombycis and evaluated their inhibitory effects on nitric oxide (NO) production as an indicator of their anti-inflammatory activities using lipopolysaccharide (LPS)-stimulated murine macrophages. Compound 4 showed the most potent inhibitory activity on NO production. It was identified as mulberrofuran K (MFK). Anti-inflammatory activity of MFK was then carried out using LPS-stimulated RAW264.7 cells. MFK suppressed the production of NO, reactive oxygen species (ROS), and proinflammatory cytokines (interleukin (IL)-1ß, IL-6 and tumor necrosis factor alpha (TNF-α)) in a concentration-dependent manner. Western blot analysis revealed that MFK treatment inhibited expression levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). MFK also inhibited transcriptional activation of nuclear factor-κB (NF-κB) and extracellular-regulated kinases (ERK) 1/2 in LPS-stimulated murine macrophages. These results suggest that MFK, an anti-inflammatory constituents of M. bombycis cortex, has potential as a therapeutic candidates for preventing and treating inflammatory diseases.

Hispidulin-7-O-Neohesperidoside from Cirsium japonicum var. ussuriense Attenuates the Production of Inflammatory Mediators in LPS-Induced Raw 264.7 Cells and HT-29 Cells.

BACKGROUND: Inflammatory bowel disease (IBD) is characterized by chronic inflammation of the gastrointestinal tract and involves secretion of inflammatory mediators. The flavone diglycoside hispidulin-7-O-neohesperidoside (HN) isolated from the methanolic extract of aerial parts of Cirsium japonicum var. ussuriense, but its pharmacologic activities, with the exception of alleviation of alcohol toxicity, have not been investigated to date. OBJECTIVE: The aim of the present study was to investigate the anti-inflammatory activities of HN for the treatment of chronic inflammatory illnesses, including IBD. MATERIALS AND METHODS: In lipopolysaccharide (LPS)-induced RAW264.7 cells and HT-29 cells, the effects of HN on cell viability and nitric oxide (NO) production were examined via MTT assay and the Griess reaction, respectively. The expression levels of interleukin (IL)-1α, IL-8, and tumor necrosis factor (TNF)-α and inducible nitric oxide synthase (iNOS) protein levels were measured by enzyme-linked immunosorbent assay and Western blotting, respectively. RESULTS: HN concentration-dependently inhibited NO production in LPS-induced RAW 264.7 cells. Treatment with HN considerably downregulated the levels of the pro-inflammatory cytokines, IL-1ß and TNF-α and the iNOS protein level in LPS-induced RAW 264.7 cells. Furthermore, HN inhibited the production of the chemotactic cytokine, IL-8, in LPS-induced HT-29 cells. CONCLUSION: HN has potential as an anti-inflammatory agent to prevent and/or treat IBD. SUMMARY: Hispidulin-7-O-neohesperidoside (HN) is flavone diglycoside isolated from the methanolic extract of aerial parts of Cirsium japonicum var. ussuriense.HN concentration-dependently inhibited NO production and considerably downregulated the levels of the proinflammatory cytokines, IL-1ß and TNF-α, and the iNOS protein level in LPS-induced RAW 264.7 cells.HN inhibited the production of the chemotactic cytokine, IL-8, in LPS-induced HT-29 cells.HN has potential as an anti-inflammatory agent to prevent and/or treat IBD. Abbreviations used: IBD: Inflammatory bowel disease, HN: hispidulin-7-O-neohesperidoside, LPS: lipopolysaccharide, NO: nitric oxide, IL: interleukin, TNF: tumor necrosis factor, iNOS: inducible nitric oxide synthase, CD: Crohn's disease, UC: ulcerative colitis, RT: room temperature, DMEM: Dulbecco's modified Eagle's medium, FBS: fetal bovine serum, PBS: phosphate buffered saline, SDS: sodium dodecyl sulfate, PVDF: polyvinylidene difluoride, SD: standard deviation.

Triterpenoids Isolated from Alnus japonica Inhibited LPS-Induced Inflammatory Mediators in HT-29 Cells and RAW264.7 Cells.

Alnus japonica (Betulaceae) is a broad-leaved tree easily found in damp regions within the mountains of Korea and Japan. Four triterpenoids (1-4) from the fruits of A. japonica, including the newly isolated 3ß-hydroxy-lanost-9(11),23(24)-dien-25,26-diol (3), inhibited the lipopolysaccharide (LPS)-induced expression of the chemotactic cytokine interleukin (IL)-8 and nitric oxide (NO) production in HT-29 colon epithelial cells and RAW264.7 macrophages, respectively. Among these triterpenoids, compound 4, which showed the most potent inhibitory activity, effectively down-regulated LPS-induced protein expression of inducible nitric oxide synthase (iNOS) in RAW264.7 cells and in HT-29 cells. Also, compound 4 concentration-dependently inhibited the levels of LPS-induced pro-inflammatory cytokines, IL-1ß and IL-6 in macrophage cells. These triterpenoids isolated from A. japonica fruits are thought to contribute to the anti-inflammatory activities of macrophage and colon epithelial cells, which are important for regulating the colon immune system. They are expected to be potential candidates for therapeutic agents against inflammatory bowel disease.

Vaccinium angustifolium Root Extract Suppresses FcɛRI Expression in Human Basophilic KU812F Cells.

Vaccinium angustifolium, commonly known as the lowbush blueberry, is a rich source of flavonoids, with which various human physiological activities have been associated. The present study focuses on the investigation of the effect of the methanolic extract of V. angustifolium root extract (VAE) on high affinity immunoglobulin E receptor (FcɛRI) α chain antibody (CRA-1)-induced allergic reaction in human basophilic KU812F cells. The total phenolic content of VAE was found to be 170±1.9 mg gallic acid equivalents/g. Flow cytometry analysis revealed that the cell surface expression of FcɛRI was suppressed in a concentration-dependent manner upon culture with VAE. Reverse-transcriptase polymerase chain reaction analysis showed that the mRNA level of the FcɛRI α chain was reduced in a concentration-dependent manner as a result of VAE treatment. Western blot analysis revealed that the protein expression of FcɛRI and the phosphorylation of extracellular signal-regulated kinases (ERK) 1/2 were concentration-dependently inhibited by VAE. We determined that VAE inhibited anti-CRA-1-induced histamine release, in addition to the elevation of intracellular calcium concentration ([Ca2+]i), in a concentration-dependent manner. These results indicate that VAE may exert an anti-allergic effect via the inhibition of calcium influx and histamine release, which occurs as a result of the down-regulation of FcɛRI expression through inhibition of ERK 1/2 activation.

Achyranthes japonica Nakai Water Extract Suppresses Binding of IgE Antibody to Cell Surface FcɛRI.

Achyranthes japonica Nakai (AJN) water extract has a variety of physiological properties, including anti-diabetic, anti-cancer, anti-inflammatory, anti-microbial, and anti-oxidative activities. In the present study, the inhibitory effects of AJN extract were investigated in high affinity immunoglobulin E receptor (FcɛRI)-mediated KU812F cells activation. AJN extract showed suppressive effects on histamine release and intracellular calcium [Ca2+]i elevation from anti-FcɛRI antibody (CRA-1)-stimulated cells in a dose-dependent manner. Flow cytometric analysis showed that AJN extract treatment caused a dose-dependent decrease in the cell surface FcɛRI expression and the binding between the cell surface FcɛRI and the IgE antibody. Moreover, reverse transcription-polymerase chain reaction analysis showed that levels of the mRNA for the FcɛRI α chain was decreased by treatment with AJN extract. These results indicate that AJN extract may exert anti-allergic effects via the inhibition of calcium influx and histamine release, which occurs as a result from the down-regulation of the binding of IgE antibody to cell surface FcɛRI. This mechanism may occur through FcɛRI expression inhibition.

6-Methoxyluteolin from Chrysanthemum zawadskii var. latilobum suppresses histamine release and calcium influx via down-regulation of FcεRI α chain expression.

Mast cells and basophils are important effector cells in immunoglobulin-E (IgE)-mediated allergic reactions. Using the human basophilic KU812F cells, we assessed the inhibitory effects of 6-methoxyluteolin, isolated from Chrysanthemum zawadskii, in the FcεRI-mediated allergic reaction. We determined that 6-methoxyluteolin inhibited anti-FcεRI α chain antibody (CRA-1)-induced histamine release, as well as elevation of intracellular calcium concentration [Ca2+]i in a dose-dependent manner. Moreover, the inhibitory effects of 6-methoxyluteolin on the cell surface expression and the mRNA level of the FcεRI α chain were determined by flow cytometric analysis and reverse transcription-polymerase chain reaction (RTPCR), respectively. Therefore, these results show that 6- methoxyluteolin is a potent inhibitor of histamine release and calcium influx via down-regulation of the FcεRI α chain.