Bioactivity-boosting strategy based on combination of anti-allergic O-methylated catechin with a Citrus flavanone, hesperetin.

Many patients with allergies have anxiety about taking anti-allergic medicines due to their side effects and increased medical expenses. Thus, developing functional foods/agricultural products for allergy prevention is strongly desired. In this study, we revealed that a Citrus flavanone, hesperetin, amplified IgE/antigen-mediated degranulation-inhibitory potency of anti-allergic catechin, (-)-epigallocatechin-3-O-(3-O-methyl) gallate (EGCG3''Me), in the rat basophilic/mast cell line RBL-2H3. Hesperetin also significantly elevated the activation of acid sphingomyelinase (ASM), essential for eliciting anti-allergic effect of EGCG3''Me through the cell surficial protein, 67-kDa laminin receptor (67LR). Furthermore, oral administration of the highly absorbent hesperidin, α-glucosyl hesperidin, also enhanced the inhibitory potency of EGCG3''Me-rich 'Benifuuki' green tea (Camellia sinensis L.) on passive cutaneous anaphylaxis (PCA) reaction evoked by IgE/antigen in BALB/c mice. These observations indicate that hesperetin amplifies the ability of EGCG3''Me to inhibit the IgE/antigen-mediated degranulation through activating ASM signaling.

The effects and the mechanisms of naringenin from Artemisia ordosica Krasch on allergic rhinitis based on mast cell degranulation model and network pharmacology.

OBJECTIVES: The ethyl acetate extraction of Artemisia ordosica Krasch (AOK) root showed anti-allergic rhinitis (AR) effect, while the active compounds and pharmacological targets were unknown. METHODS: The P815 degranulation was established by cell counting kit 8 assay, ß-hexosaminidase releasing assay and toluidine blue staining. The flavonoids were screened in vitro. Then toluidine blue staining and ELISA were carried out to investigate the anti-inflammatory effects of the active compound. Network pharmacology was implemented to explain the mechanisms of the active compound. iGEMDOCK was used to investigate the binding between active compound and hub targets. KEY FINDINGS: C48/80 was the optimum reagent in triggering P815 degranulation. Naringenin could significantly decrease P815 degranulation. Meanwhile, naringenin could remarkably increase the IL-4 and decrease the tumour necrosis factor-α. The effect of naringenin on AR was achieved by regulating multiple targets (e.g. AKT1, MAPK3, VEGFA) and pathways (e.g. pathways in cancer, VEGF signalling pathway). Nine hub proteins were obtained by topological analysis. Multiple hydrogen bonds and van der Waals forces were formed between the naringenin and the residues of hub proteins. CONCLUSIONS: Naringenin might be one of the effective ingredients of AOK against AR. And its effects could achieve through regulating multiple targets and pathways.

Lactic Acid Bacteria Fermented Cordyceps militaris (GRC-SC11) Suppresses IgE Mediated Mast Cell Activation and Type I Hypersensitive Allergic Murine Model.

Cordyceps militaris (C. militaris) has various biomedical applications in traditional oriental medicine for different diseases including inflammatory and immune-dysregulated diseases. It is a reservoir of nutritional components such as cordycepin, polysaccharides, and antioxidants. To improve its bioactivity, we fermented C. militaris with a Pediococcus pentosaceus strain isolated from a salted small octopus (SC11). The current study aimed to evaluate whether P. pentosaceus (SC11) fermentation could enhance the anti-allergic potential of C. militaris cultured on germinated Rhynchosia nulubilis (GRC) against a type I hypersensitive reaction in in vitro and in vivo studies. Total antioxidant capacity and cordycepin content were significantly increased in GRC after SC11 fermentation. GRC-SC11 showed significantly enhanced anti-allergic responses by inhibiting immunoglobulin E (IgE)/antigen-induced degranulation in RBL-2H3 cells, compared to GRC. The results demonstrated the significant inhibition of phosphorylated spleen tyrosine kinase (Syk)/ p38/GRB2-associated binding protein 2 (Gab2)/c-jun in IgE/Ag-triggered RBL-2H3 cells. Furthermore, suppressed mRNA levels of interleukin-4 (IL-4) and tumor necrosis factor-α (TNF-α) in IgE/Ag-activated RBL-2H3 cells were observed. GRC-SC11 significantly ameliorated IgE-induced allergic reactions by suppressing the ear swelling, vascular permeability, and inflammatory cell infiltration in passive cutaneous anaphylaxis (PCA) BALB/c mice. In conclusion, GRC fermented with P.pentosaceus exerted enhanced anti-allergic effects, and increased the cordycepin content and antioxidants potential compared to GRC. It can be used as bio-functional food in the prevention and management of type I allergic diseases.

Anti-Allergic Effects of the Subcritical Water Extract Powder of Citrus unshiu in Mouse and Guinea Pig Models.

We evaluated the effects of unripe mandarin orange (Citrus unshiu) extract powder (unripe mandarin extract powder [UMEP]) treated with subcritical water on allergic diseases by using animal models. High performance liquid chromatography (HPLC) analysis revealed that subcritical water is a more effective solvent than alcohol and hot water, as it quickly extracted approximately 90% of the functional compounds narirutin (1) and hesperidin (2) from whole fruits. Repeated oral administration of UMEP significantly reversed the peripheral blood flow decline observed during the promotion of allergies after sensitization with the antigen, hen egg-white lysozyme (HEL). UMEP also significantly inhibited compound 48/80-induced scratching behavior in HEL-sensitized mice, which are more sensitive to itching stimuli than are normal mice, without suppressing locomotor activity. In addition, repeated oral administration of UMEP in ovalbumin-challenged guinea pigs significantly suppressed the late phase of nasal airway resistance. This study provides evidence that the subcritical water extract powder of unripe C. unshiu fruit is an effective anti-allergic functional food.

Inhibitory effects of cyanidin-3-O-glucoside in black soybean hull extract on RBL-2H3 cells degranulation and passive cutaneous anaphylaxis reaction in mice.

Black soybean hull extract (BSHE) exhibits a variety of biological activities. However, little is known about the effects of BSHE on immunoglobulin E (IgE)-mediated type I allergic reactions. The anti-allergic effect of BSHE was assessed with the degranulation assay using rat basophilic leukemia RBL-2H3 cells and the passive cutaneous anaphylaxis (PCA) reaction in mice. An active compound in BSHE was identified by ultra-performance liquid chromatography coupled to diode array detection and electrospray ionization tandem mass spectrometry analysis. BSHE inhibited the release of ß-hexosaminidase and histamine in RBL-2H3 cells, and cyanidin-3-O-glucoside (C3G) was identified as one of its active compounds. Oral administering of 200 µmol/kg of C3G to IgE-sensitized mice prior to antigen injection suppressed the PCA reaction, as compared with control (p < 0.01). Intravenous administration of BSHE (C3G content, 5.4%) more strongly inhibited PCA responses at lower doses (100 mg/kg, p < 0.01) than oral administration (1,000 mg/kg, p = 0.059). Intravenous C3G also suppressed PCA response at a low dose (40 mg/kg, p < 0.05), showing the same trend as BSHE. This information can be useful to design appropriate formulations of anthocyanin-based drug products to suppress allergic reactions. This study provides evidence for the potential use of BSHE and C3G for the prevention or the treatment of type I allergies.

Inhibitory Effects of Green Asparagus Extract, Especially Phospholipids, on Allergic Responses in Vitro and in Vivo.

Asparagus (Asparagus officinalis L.) is one of the widely consumed vegetables. To investigate the mechanism underlying the anti-allergic responses of asparagus, we extracted different fractions from asparagus and measured their inhibitory effects on ß-hexosaminidase release in RBL-2H3 cells in vitro and an atopic dermatitis NC/Nga mouse model in vivo. The lipid fractions from asparagus were extracted with 50% ethanol, separated using chloroform by liquid-liquid phase separation, and fractionated by solid-phase extraction. Among them, acetone fraction (rich in glycolipid) and MeOH fraction (rich in phospholipid) markedly inhibited ß-hexosaminidase release from RBL-2H3 cells. In NC/Nga mice treated with picryl chloride, atopic dermatitis was alleviated following exposure to the 50% EtOH extract, acetone fraction, and methanol fraction. The inhibitory effects of asparagus fractions in vivo were supported by the significant decrease in serum immunoglobulin E (IgE) levels. The phospholipid fractions showed significantly better inhibitory effects, and phosphatidic acid from this fraction showed the best inhibitory effect on ß-hexosaminidase release. In mice challenged with ovalbumin (OVA), oral administration of asparagus extract and its fractions decreased the OVA-specific IgE level and total IgE, indicating that these effects may be partly mediated through the downregulation of antigen-specific IgE production. Taken together, the present study shows for the first time that asparagus extract and its lipid fractions could potentially mitigate allergic reactions by decreasing degranulation in granulocytes. Our study provides useful information to develop nutraceuticals and functional foods fortified with asparagus.

Antioxidant compounds of Petasites japonicus and their preventive effects in chronic diseases: a review.

Petasites japonicus (P. japonicus) is a plant of the Asteraceae family. Its roots and stems have been used for the treatment or the prophylaxis of migraine and tension headache as a traditional Chinese medicine in Japan and Korea. Sesquiterpenoids, lignans, and flavonoids are components of P. japonicus. Regarding the biological activity of P. japonicus, its anti-allergic effect has been researched extensively using IgE antigen-stimulated degranulation of RBL-2H3 cells or passive cutaneous anaphylaxis reaction in experimental animal models. The study of the antioxidant activity of P. japonicus was initiated approximately 15 years ago using in vitro assays. In addition, its in vivo effect has also been examined in animal models with induced oxidative injury. Moreover, recently, many types of antioxidant compounds have been rapidly and simultaneously identified using the liquid chromatography-mass spectrometry technique. The number of reports on the other functions of this plant, such as its neuroprotective and anti-inflammatory effects, has been increasing. In this review, I summarized the studies of functional foods derived from P. japonicus, which may provide a basis for the development of potential functional foods. Finally, I discuss the future research avenues in this field.

Potential in vitro anti-allergic, anti-inflammatory and cytotoxic activities of ethanolic extract of Baliospermum montanum root, its major components and a validated HPLC method.

BACKGROUND: The root of Baliospermum montanum has been used as an ingredient of traditional Thai medicines for the treatments of several diseases including itching eczema, muscle and joint inflammation, and cancer. Few studies have been done on phytochemical components of this root. In this study, we isolated major compounds of the crude ethanolic extract of B. montanum root and developed and validated a high performance liquid chromatographic (HPLC) method for the determination of its major components. We then investigated anti-allergic, anti-inflammatory and cytotoxic activities of the extract. METHODS: The aims of this study were to investigate in vitro activities including inhibitory effect of ß-hexosaminidase released from RBL-2H3 cells, inhibition of nitric oxide (NO) production from RAW 264.7 cells and cytotoxic activity against cancerous liver cell lines (HepG2 and KKU M156) by using sulforhodamine B (SRB) assay. Isolation of major components was conducted by using column chromatographic method. Isolated major compounds were analyzed by using high performance liquid chromatography (HPLC). RESULTS: The crude extract exhibited the highest cytotoxic activity, with IC50 less than 1 µg/mL, while its anti-allergy and anti-inflammation were also potent with IC50 less than 6 µg/mL. Three propiophenones isolated from B. montanum root exhibited moderate cytotoxic activities (IC50 > 20 µg/mL). Two of the propiophenones found were major components that can be detected by HPLC. The developed and validated HPLC method showed good accuracy, precision, and linearity. CONCLUSION: The results of this study suggested that ethanolic extract of of B.montanum root can be a potential source of anti-allergy, anti-inflammation, and anti-cancer compounds. The isolated compounds can serve as markers when B. montanum is used in herbal remedies but not as overall responsive markers. The HPLC method developed may be useful for quality control in the production of the extract and for further formulation developments. However, investigation of several associated biological activities is necessary before the development can proceed further. Minor active compounds should be isolated and a more sensitive analytical method should be developed to detail the key responsive components of the ethanolic extract of B. montanum root.

Identification of degranulation inhibitors from rooibos (Aspalathus linearis) tea in rat basophilic leukaemia cells.

Quercetin, luteolin and chrysoeriol were identified from rooibos tea as degranulation inhibitors in rat basophilic leukaemia cells. The degranulation inhibitory activity of chrysoeriol was first discovered in the present study. When quercetin, luteolin and chrysoeriol were mixed in the ratio that occurs in rooibos tea extract, the mixture inhibited antigen- and calcium ionophore-stimulated degranulation to the same degree as that by the whole rooibos tea extract. These findings indicate that these three flavonoids are the key factors underlying the degranulation inhibitory activity of rooibos tea.

Anti-allergy Effect of Gouminjian and Its Components / 中国实验方剂学杂志

Guominjian is a prescription created by famous traditional Chinese medicine (TCM) doctor ZHU Zhan-yu. The prescription is composed of four TCM:Saposhnikoviae Radix, Mume Fructus, Schisandrae Chinensis Fructus and Stellariae Radix, which are commonly used in clinical practice for allergic skin diseases, such as urticaria, atopic dermatitis and eczema. Although the medicinal composition is simple, the curative effect is remarkable, and the total effective rate of treatment in clinic is as high as 90%. Guominjian has the effects on phlegm and blood stasis, and can relieve itching and reduce capillary permeability by antagonizing histamine, reducing serum immune globulin (Ig) E, inhibiting mast cell degranulation and changing the differentiation of CD4+ T cell subsets, with anti-allergy effect at multi-target, multi-path, multi-levels. Domestic and foreign scholars have screened out the chemical constituents of single herbs in Guominjian, and found anti-allergic active ingredients, such as α-cubebenoate, cimicin, schisandrin, and saposhnikovia divaricata polysaccharide, and the mechanisms are different. However, the effects of TCM compounds are not equal to the simple combination of single herbs, and it is not possible to measure the efficacy of an active ingredient in a single herb. The anti-allergic chemical composition in Guominjian should be comprehensively analyzed to study the anti-allergy effect of Guominjian in an all-round way. However, the current research on Guominjian is only limited to the pharmacodynamics, but there is still lack systematic, comprehensive and holistic understanding of anti-allergic active components in Guominjian, which limits the secondary development and the establishment of standards of Guominjian. Therefore, the authors systematically collect and summarize the anti-allergic modern research results of Guominjian and its components by searching domestic and foreign literatures, in order to further define the research and new mechanism of anti-allergic chemical components and mechanism of Guominjian, and provide reference for the development of anti-allergic new drugs.

Saccharopolytide A, a new cyclic tetrapeptide with rare 4-hydroxy-proline moieties from the deep-sea derived actinomycete Saccharopolyspora cebuensis MCCC 1A09850.

A genome mining analysis on the deep-sea derived actinomycete Saccharopolyspora cebuensis MCCC 1A09850 indicated its potential to produce polypeptides. Accordingly, a systematic chemical investigation was conducted, which resulted in the isolation of one new cyclic tetrapeptide (saccharopolytide A, 1) and two known polyketides (2, 3) along with six other miscellaneous compounds (4‒9). Mainly by analysis of the 1D, 2D NMR and MS data, the chemical structure of saccharopolytide A was established as cyclo-(l-Leu-4-hydroxy-l-Pro-l-Phe-4-hydroxy-l-Pro). All isolates were evaluated for anti-allergic and anti-tumor bioactivities. Indol-3-carbaldehyde (4) showed weak anti-allergic effect with IC50 value of 55.75 µg/mL. And 2 showed weak anti-proliferative activity against Hela and H1299 tumor cell lines. Our results consolidate the potential of deep-sea-derived microorganisms to produce structurally interesting compounds.

The Potential of Triterpenoids from Loquat Leaves (Eriobotrya japonica) for Prevention and Treatment of Skin Disorder.

The leaves of loquat (Eriobotrya japonica) possess high medicinal value and have been used as traditional medicines. However, there are no evidence-based studies on the skin-care effects of E. japonica leaves. To explore new biological activities of E. japonica leaves against skin disorder and to gain a better understanding of the chemical components associated with bioactivities, we evaluated 18 triterpenoids from E. japonica leaves on anti-melanogenesis, anti-acne, anti-allergy and anti-aging activities. Our results revealed that eight compounds showed anti-melanogenesis activity, of which ursolic acid (1) and maslinic acid (7) were the most potent with the similar selective index to that of arbutin. Structure-activity relationship and possible mechanism of active compounds were proposed. Twelve compounds exhibited anti-acne effect; ursolic acid (1), maslinic acid (7), corosolic acid (8) and euscaphic acid (12) showed highest activities against P. acnes. Four compounds displayed anti-allergy and anti-inflammatory activity; 3-epicorosolic acid (9) and euscaphic acid (12) showed marked activity against ß-hexosaminidase release. Finally, ursolic acid (1), pomolic acid (10), colosolic acid (8) and its methylated derivative (6) exhibited the highest anti-aging activity by stimulating collagen and hyaluronic acid (HA) production. Our findings provide valuable evidence that E. japonica leaves have potential applications as ingredients of function foods or cosmetics for health benefits and a number of triterpenoids may play an important role in these bioactivities.

The anti-allergic and anti-inflammatory effects of Benjakul extract (a Thai traditional medicine), its constituent plants and its some pure constituents using in vitro experiments.

Benjakul (BJK), a Thai traditional medicine preparation, has long been used for balanced health, controlled abnormal of element in the body, carminative, and relief of flatulence. It is composed of five plants: Piper interruptum Opiz., Piper longum L., Piper sarmentosum Roxb., Plumbago indica L., and Zingiber officinale Roscoe. The ethanolic extracts of BJK, its five individual plants, and pure constituents of BJK were investigated for their anti-allergic activity using immunoglobulin E (IgE)-sensitized ß-hexosaminidase in the rat basophilic leukemia-2H3 (RBL-2H3) cells and anti-inflammatory activity using lipopolysaccharide (LPS)-induced nitric oxide (NO) and tumor necrosis factor-alpha (TNF-α) in the murine macrophage (RAW 264.7) cells. The ethanolic extracts of BJK showed anti-allergic activity (IC50=12.69µg/ml) and exhibited potent NO inhibitory effect (IC50=16.60µg/ml), but inactive on TNF-α release. Moreover, 6-shogaol and plumbagin, two pure compounds from BJK, showed higher anti-allergic activity than the ethanolic BJK extract with IC50 values of 0.28 and 4.03µg/ml, respectively. These compounds were significantly higher than chlorpheniramine (CPM), standard drug, with IC50 value of 17.98µg/ml. Determination of the anti-inflammatory activity by measuring the inhibition of NO production presented that plumbagin and 6-shogaol exhibited higher than crude BJK extract with IC50 values of 0.002 and 0.92µg/ml, respectively. In particular, plumbagin also showed higher anti-inflammatory than prednisolone, positive control, with IC50 value of 0.59µg/ml. 6-Shogaol also showed inhibitory effect on TNF-α release (IC50=9.16µg/ml). These preliminary results may provide some scientific support for the use of BJK for the anti-allergic treatment and inflammatory disorders through the inhibition of NO production.

Inhibitory effects of norlignans isolated from Anemarrhena asphodeloides on degranulation of rat basophilic leukemia- 2H3Cells.

Anemarrhena asphodeloides is known to suppress inflammation and lower various fevers. To determine the active component of A. asphodeloides, ethanol (EtOH) extract of A. asphodeloides rhizomes was fractionized. The compounds isolated from the dichloromethane (CH2Cl2) soluble fraction were identified as 4'-O-methylnyasol (1), nyasol (2), 3″-methoxynyasol (3), 3″-hydroxy-4″-methoxy-4″-dehydroxynyasol (4), 4-hydroxybenzaldehyde (5), and 4-hydroxyacetophenone (6). The four norlignans (1-4) potently inhibited the release of ß-hexosaminidase from immunoglobulin E (IgE)/dinitrophenol-conjugated bovine serum albumin (DNP-BSA)-treated rat basophilic leukemia (RBL)-2H3 and A23187 plus phorbol 12-myristate 13-acetate co-treated isolated rat primary mast cells, as markers of degranulation and histamine release. The intraperitoneal treatment with the EtOH extract significantly suppressed the fetal reaction, and serum histamine release induced by compound 48/80 in mice. These results suggest that the four active norlignan compounds and the EtOH extract of A. asphodeloides may have potential to be developed as medicines for the treatment of allergies by inhibiting the activation of mast cells.

Anti-Allergic Effect of Oroxylin A from Oroxylum indicum Using in vivo and in vitro Experiments.

Oroxylum indicum has long been used in Asian traditional medicine to prevent and treat respiratory diseases, diabetes, diarrhea and other conditions. Oroxylin A is a flavone that is present in Oroxylum indicum and in Scutellaria baicalensis. Because the root extracts of both plants have been shown to have anti-allergic effects, the authors investigated whether oroxylin A is likely to have beneficial effects on allergic asthma using female Balb/c mice and rat RBL-2H3 mast cells. Antigen-induced degranulation was measured in vitro by measuring ß-hexosaminidase activity. A murine ovalbumin-induced allergic asthma model was used to test the in vivo efficacy of oroxylin A. Sensitization and challenge of ovalbumin induced allergic asthma responses, the accumulations of eosinophils and Th2 cytokine levels in bronchoalveolar lavage fluid and lung tissues. Oroxylin A administration decreased numbers of inflammatory cells, especially eosinophils, and reduced the expression and secretion of Th2 cytokines, including IL-4 and IL-13, in lung tissues and bronchoalveolar lavage fluid. Histologic studies showed oroxylin A reduced inflammatory signs and mucin production in lungs. These findings provide evidence that oroxylin A has potential use as an anti-allergic therapeutic.

Petatewalide B, a novel compound from Petasites japonicus with anti-allergic activity.

ETHNOPHARMACOLOGICAL RELEVANCE: The giant butterbur Petasites japonicus is used to treat asthma and allergic diseases in traditional Korean, Japanese, and Chinese medicine. AIM OF THE STUDY: To elucidate the anti-allergic effect of Petasites genus, we studied effects of several compounds from Petasites japonicus leaves and found a novel bakkenolide-type sesquiterpine. In the present study, anti-allergic and anti-inflammatory effects of the new compound was examined using in vivo and in vitro experiments. MATERIALS AND METHODS: The novel compound was isolated from Petasites japonicus leaves and named petatewalide B. Antigen-induced degranulation and Ca(2+) mobilization were measured in RBL-2H3 mast cells by measuring ß-hexosaminidase activity and fluorescence change of Ca(2+) probe, fura-2. Induction of inducible nitric oxide synthase and cyclooxygenase 2 was measured by Western blotting in peritoneal macrophages. In addition, ovalbumin-induced asthma model was used for in vivo efficacy test of petatewalide B. Membrane potential was estimated by measuring fluorescence change of DiBAC in C6 glioma cells. RESULTS: Petatewalide B inhibited the antigen-induced degranulation of ß-hexosaminidase in RBL-2H3 mast cells, but did not affect antigen-induced Ca(2+) increase in the cells. Petatewalide B also showed inhibition of the LPS-induced induction of iNOS, but not COX-2 in mouse peritoneal macrophages. Nitric oxide production was also inhibited by petatewalide B in macrophages. In the ovalbumin-induced asthma model, petatewalide B strongly inhibited accumulations of eosinophils, macrophages, and lymphocytes in bronchoalveolar lavage fluid. Petatewalide B increased the membrane potential of C6 glioma cells in a concentration-dependent manner. CONCLUSION: Petatewalide B from Petasites genus not only has anti-allergic and anti-inflammatory effects but also induces a transient increase of membrane potential in C6 glioma cells.

Anti-Allergic Effect of Oroxylin A from Oroxylum indicum Using in vivo and in vitro Experiments

Oroxylum indicum has long been used in Asian traditional medicine to prevent and treat respiratory diseases, diabetes, diarrhea and other conditions. Oroxylin A is a flavone that is present in Oroxylum indicum and in Scutellaria baicalensis. Because the root extracts of both plants have been shown to have anti-allergic effects, the authors investigated whether oroxylin A is likely to have beneficial effects on allergic asthma using female Balb/c mice and rat RBL-2H3 mast cells. Antigen-induced degranulation was measured in vitro by measuring β-hexosaminidase activity. A murine ovalbumin-induced allergic asthma model was used to test the in vivo efficacy of oroxylin A. Sensitization and challenge of ovalbumin induced allergic asthma responses, the accumulations of eosinophils and Th2 cytokine levels in bronchoalveolar lavage fluid and lung tissues. Oroxylin A administration decreased numbers of inflammatory cells, especially eosinophils, and reduced the expression and secretion of Th2 cytokines, including IL-4 and IL-13, in lung tissues and bronchoalveolar lavage fluid. Histologic studies showed oroxylin A reduced inflammatory signs and mucin production in lungs. These findings provide evidence that oroxylin A has potential use as an anti-allergic therapeutic.

Anti-allergic effect of α-cubebenoate isolated from Schisandra chinensis using in vivo and in vitro experiments.

AIM OF THE STUDY: In Oriental countries, the dried fruits of Schisandra chinensis are extensively used in traditional medicine to treat asthma, gonorrhea, and other diseases. Recently, α-cubebenoate was isolated as an anti-inflammatory component from Schisandra chinensis. In the present study, the authors examined the anti-allergic effect of α-cubebenoate using in vivo and in vitro experiments. MATERIALS AND METHODS: α-Cubebenoate was isolated from an extract of Schisandra chinensis fruits. Antigen-induced degranulation and Ca(2+) mobilization were measured in RBL-2H3 mast cells. In addition, BALB/c mice were sensitized with ovalbumin and aluminum hydroxide, and then challenged with ovalbumin for three consecutive days. α-Cubebenoate (1mg/kg) was administered intraperitoneally 30min before each ovalbumin challenge. RESULTS: In RBL-2H3 mast cells, α-cubebenoate inhibited antigen-induced degranulation and increase of intracellular Ca(2+) concentrations. In the ovalbumin-induced asthma model, α-cubebenoate suppressed bronchiolar structural changes induced by ovalbumin challenge. Furthermore, α-cubebenoate strongly inhibited accumulations of eosinophils, macrophages, and lymphocytes in bronchoalveolar lavage fluid. α-Cubebenoate also suppressed Th2 cytokines (IL-4 and IL-13) and TGF-ß1 in lung tissues and in immune cells at the mRNA and protein levels. CONCLUSION: α-Cubebenoate has an inhibitory effect on allergic inflammation and could be utilized as an agent for the treatment of asthma.

Effects of the Subaleurone Layer of Rice on Macrophage Activation and Protection of Pollen Allergy in a Murine Model.

BACKGROUND/AIM: Oral intake of lipopolysaccharide (LPS) has been demonstrated to be effective in the prevention of various diseases. We have found that the subaleurone layer of rice contains a large amount of LPS. The aim of this study was to evaluate the role of this layer in innate immunity. MATERIALS AND METHODS: Using the Saika-style rice polishing process, a sbaleurone layer and the rice retaining a subaleurone layer and polished white rice were prepared from brown rice. Using hot-water extracts from rice, LPS content was measured by the Limulus reaction and the effect of activation of macrophages was evaluated on the basis of their phagocytic activity and nitric monoxide (NO) and tumor necrosis factor (TNF) production levels. Toll-like receptor (TLR)-2-, TLR-4- and TLR-9-transfected human embryonic kidney (HEK) cells were used to identify the activation pathway. An allergy mouse model was used to evaluate the prevention of pollen allergy. RESULTS: When compared to polished white rice, rice retaining a subaleurone layer had a 6-fold increase in LPS and an increased macrophage activation when phagocytic activity and NO and TNF production were used as indices. TRL4 was the major pathway for such activation. Anti-allergy test by oral intake of subaleurone showed a significant preventive effect for pollen allergy. CONCLUSION: Compared to polished white rice, rice retaining a subaleurone layer contained a high level of LPS with higher macrophage activation. Furthermore, oral administration of the rice demonstrated a preventive effect for pollen allergy, thus indicating its utility as a functional food that has a regulatory effect on innate immunity.

Oral Administration of Lipopolysaccharide of Acetic Acid Bacteria Protects Pollen Allergy in a Murine Model.

BACKGROUND/AIM: Lipopolysaccharide (LPS), a major component of the cell wall of Gram-negative bacteria, is known to possess strong immune-regulatory activity. We have found and reported the existence of biologically-active LPS in acetic acid bacteria. The LPS shows Limulus-positive activity and activation of macrophages to produce nitric oxide and tumor necrosis factor. In this study, we investigated the anti-allergic effect of an orally-administrated acetic acid bacteria extract containing LPS; the cedar pollinosis model was used. MATERIALS AND METHODS: Acetic acid bacteria were isolated from various fruits by Nodai kaihen medium. Then, the anti-allergic effect of acetic acid bacteria extracts was investigated. BALB/c mice were immunized with a mixture of cedar pollen and alum into their peritoneal cavity; they also received additional immunizations of pollen to nasal cavity. After immunizing the mice with pollen into their nasal cavity to trigger an allergic reaction, the frequency of nose scratching was counted for 5 min. RESULTS: The bacteria were cultured and prepared and the water-extract contained about 1-10 mg/ml of Limulus positive substances. The extract of acetic acid bacteria induced higher levels of interleukin (IL)-10 and FOXP3 mRNA expression in macrophages (RAW246.7 cell), as assessed by DNA microarray analysis. Oral administration of the acetic acid bacteria extract demonstrated significantly less scratching numbers than control water group with pollen immunization. CONCLUSION: These results showed that LPS in acetic acid bacteria has the potential to protect from an allergic reaction, especially from cedar pollinosis.