Changda Liu

Berberine and limonin suppress IgE production by human B cells and peripheral blood mononuclear cells from food-allergic patients.

BACKGROUND Currently, there is no satisfactory treatment for IgE-mediated food allergy. Food Allergy Herbal Formula 2 (FAHF-2) and butanol-purified FAHF-2 (B-FAHF-2) have been shown to protect against peanut-induced anaphylaxis and inhibit IgE synthesis in a murine model. OBJECTIVE To determine which herbs and compounds in FAHF-2 and B-FAHF-2 suppress IgE production. METHODS The effect of FAHF-2 and B-FAHF-2 on IgE production was determined using a human B-cell line (U266). Individual compounds were isolated and identified using column chromatography, liquid chromatographic mass spectrometry, and nuclear magnetic resonance techniques. The potency of compounds on IgE suppression were investigated using U266 cells and verified using human peripheral blood mononuclear cells (n = 25) from peanut-allergic patients. Epsilon germline transcript expression was determined. Phosphorylated IκBα level was analyzed using the In-Cell Western assay. The mRNA expression of signal transducer and activator of transcription-3, T-box transcription factor TBX21, interferon-γ, forkhead box P3, GATA-binding protein 3, interleukin-10, and interleukin-5 also were analyzed using real-time polymerase chain reaction. RESULTS FAHF-2 and B-FAHF-2 inhibited IgE production by U266 cells. B-FAHF-2 was 9 times more effective than FAHF-2. Two compounds that inhibited IgE production were isolated from Philodendron chinensis and identified as berberine and limonin. Berberine was more potent and inhibited IgE production by peripheral blood mononuclear cells by 80% at 0.62 μg/mL. Berberine significantly inhibited ε-germline transcript expression by peripheral blood mononuclear cells. Phosphorylated IκBα level was significantly suppressed and mRNA expressions of T-box transcription factor TBX21 and signal transducer and activator of transcription-3 were significantly increased by berberine. CONCLUSION Berberine and limonin mediated IgE suppression. The mechanism by which berberine modulates ε-germline transcript expression might be through regulating the phosphorylated IκBα level and the expressions of signal transducer and activator of transcription-3 and T-box transcription factor TBX21. TRIAL REGISTRATION Clinicaltrials.gov identifier NCT00602160.

Ganoderic acid C1 isolated from the anti-asthma formula, ASHMI™ suppresses TNF-α production by mouse macrophages and peripheral blood mononuclear cells from asthma patients.

Asthma is a heterogeneous airway inflammatory disease, which is associated with Th2 cytokine-driven inflammation and non-Th2, TNF-α mediated inflammation. Unlike Th2 mediated inflammation, TNF-α mediated asthma inflammation is generally insensitive to inhaled corticosteroids (ICS). ASHMITM, aqueous extract of three medicinal herbs-Ganoderma lucidum (G. lucidum), Sophora flavescens Ait (S. flavescens) and Glycyrrhiza uralensis Fischer (G. uralensis), showed a high safety profile and was clinically beneficial in asthma patients. It also suppresses both Th2 and TNF-α associated inflammation in murine asthma models. We previously determined that G. uralensis flavonoids are the key active compounds responsible for ASHMITM suppression of Th2 mediated inflammation. Until now, there are limited studies on anti-TNF-α compounds presented in ASHMITM. The objective of this study was to isolate and identify TNF-α inhibitory compounds in ASHMITM. Here we report that G. lucidum, but not the other two herbal extracts, S. flavescens or G. uralensis inhibited TNF-α production by murine macrophages; and that the methylene chloride (MC)-triterpenoid-enriched fraction, but not the polysaccharide-enriched fraction, contained the inhibitory compounds. Of the 15 triterpenoids isolated from the MC fraction, only ganoderic acid C1 (GAC1) significantly reduced TNF-α production by murine macrophages (RAW 264.7 cells) and peripheral blood mononuclear cells (PBMCs) from asthma patients. Inhibition was associated with down-regulation of NF-κB expression, and partial suppression of MAPK and AP-1 signaling pathways. Ganoderic acid C1 may have potential for treating TNF-α mediated inflammation in asthma and other inflammatory diseases.

Effect of Antiasthma Simplified Herbal Medicine Intervention on neutrophil predominant airway inflammation in a ragweed sensitized murine asthma model

BACKGROUND Neutrophil-predominant asthma is less responsive to steroids and associated with poorer disease control. The effects of Antiasthma Simplified Herbal Medicine Intervention (ASHMI), a traditional Chinese medicine formula reported to be efficacious in asthmatic patients and murine asthma models, on neutrophil predominant asthma are unknown. OBJECTIVE To determine the effects of standard ASHMI and refined formula ASHMI (ASHMI(II)) in a neutrophil-predominant murine model of ragweed (RW) asthma and explore underlying mechanisms. METHODS BALB/c mice were systemically sensitized, intranasally challenged with RW extract, and orally treated with ASHMI, ASHMI(II), or vehicle (water). In a separate experiment, some RW sensitized mice were treated with dexamethasone before challenge. After RW challenge, airway hyperreactivity (AHR), total and differential bronchoalveolar lavage fluid leukocyte counts, lung histologic features, and bronchoalveolar lavage fluid cytokine and chemokine levels were assessed. RW stimulation of the murine macrophage cell line RAW264.7 was used to determine effects of ASHMI active compound ganoderic acid C1 (GAC1) on tumor necrosis factor α (TNF-α) production and regulation of phosphorylated IκB and histone deacetylase 2 (HDAC2) levels. RESULTS ASHMI and ASHMI(II) markedly reduced AHR, mucous production, neutrophilic inflammation, and TNF-α, interleukin 8, and interleukin 17 levels and decreased eosinophilic inflammation and TH2 responses in vivo (P < .01-.001 for all). GAC1 inhibited TNF-α production in RW-stimulated RAW264.7 cells in association with suppression of phosphorylated IκB and increased HDAC2 expression. Dexamethasone failed to reduce AHR and neutrophilic inflammation. CONCLUSION ASHMI treatment was efficacious in a murine model of neutrophil-predominant asthma via modulation of innate chemokines, TH2 responses, nuclear factor-κB, and HDAC2. ASHMI, and/or its constituent GAC1, may be a valuable option for treating neutrophil-predominant asthma.

Anti-inflammatory Effects of Ganoderma lucidum Triterpenoid in Human Crohn's Disease Associated with Downregulation of NF-κB Signaling.

Background:Crohns disease (CD) is a chronic inflammatory disease of the gastrointestinal tract. Current medications have potentially serious side effects. Hence, there is increasing interest in alternative therapies. We previously demonstrated the anti-inflammatory effects of Food Allergy Herbal Formula-2 in vitro on peripheral blood mononuclear cells (PBMCs) and mucosa from CD subjects. Here, we investigated the anti-inflammatory effects of a bioactive compound isolated from Ganoderma lucidum (G. lucidum), a key herbal constituent of Food Allergy Herbal Formula-2, in CD in vitro. Methods:Triterpene ganoderic acid C1 (GAC1) was isolated from G. lucidum. Stimulated RAW 264.7 macrophages were treated with GAC1. Human PBMCs and colonic biopsies were obtained from children with CD and cultured with or without GAC1. TNF-&agr; and other proinflammatory cytokine levels were measured in the culture supernatant. NF-&kgr;B signaling was investigated in PBMCs and colonic mucosa treated with GAC1 by In-Cell Western and Western blot analysis. Results:GAC1 decreased TNF-&agr; production by macrophages and PBMCs from CD subjects. GAC1 significantly decreased TNF-&agr;, IFN-&ggr;, and IL-17A production by inflamed colonic biopsies from CD subjects. These effects were due to downregulation of the NF-&kgr;B signaling pathway. Conclusions:GAC1 inhibited production of TNF-&agr; and other proinflammatory cytokines by PBMCs and inflamed CD colonic mucosa due to blockage of NF-&kgr;B activation. GAC1 is a key beneficial constituent in G. lucidum and the Food Allergy Herbal Formula-2 in suppressing the inflammatory cytokines found in CD and warrants clinical investigation for the treatment of CD.

The Flavonoid 7,4′‐Dihydroxyflavone Inhibits MUC5AC Gene Expression, Production, and Secretion via Regulation of NF‐κB, STAT6, and HDAC2

Mucus overproduction is a significant component of the pathophysiology of obstructive lung diseases. Currently, there are only a few medications available that inhibit mucus production. Previous studies showed that glycyrrhizin, a triterpenoid in Glycyrrhiza uralensis inhibits mucin 5AC (MUC5AC) mRNA and protein expression. Other potential mucus production inhibitory compounds contained within in G. uralensis have not been fully investigated. The aim of the present study was to determine if the G. uralensis flavonoid 7,4′‐dihydroxyflavone (7,4′‐DHF) inhibits MUC5AC gene expression, mucus production, and secretion, and if so, to elucidate the mechanism of this inhibition. 7,4′‐Dihydroxyflavone significantly decreased phorbol 12‐myristate 13‐acetate‐stimulated NCI‐H292 human airway epithelial cell MUC5AC gene expression and mucus production, at a 28‐fold lower concentration than glycyrrhizin (The half maximal inhibitory concentration IC50 value of 1.4 μM vs 38 μM, respectively); 7,4′‐DHF also inhibited MUC5AC mucus secretion. Inhibition was associated with the suppression of nuclear factor kappa‐light‐chain‐enhancer of activated B cells (NF‐κB), signal transducer and activator of transcription 6 (STAT6) activation, and enhanced histone deacetylase 2 (HDAC2) expression. In a murine model of asthma, 7,4′‐DHF‐treated mice exhibited a marked reduction in MUC5AC secretion in the bronchoalveolar lavage fluid compared with control mice. These findings, together with previous findings linking NF‐κB, STAT6, and HDAC2 modulation to the control of MUC5AC expression, demonstrate that 7,4′‐DHF is a newly identified component of G. uralensis that regulates MUC5AC expression and secretion via regulation of NF‐κB, STAT6, and HDAC2. Copyright

B-FAHF-2 plus oral immunotherapy (OIT) is safer and more effective than OIT alone in a murine model of concurrent peanut/tree nut allergy

Concurrent sensitization to peanut (PN) and tree nuts (TN), the most dangerous food allergies, is common. Current oral immunotherapy (OIT) is not fully satisfactory.

Berberine as a chemical and pharmacokinetic marker of the butanol-extracted Food Allergy Herbal Formula-2

Rationale: Food Allergy Herbal Formula‐2 (FAHF‐2) provided protection against peanut anaphylaxis in a murine model and induced beneficial immune‐modulation in humans. Butanol‐refined FAHF‐2, B‐FAHF‐2, retained safety and efficacy in the peanut allergic murine model at only 1/5 of FAHF‐2 dosage. One compound, berberine, was isolated and identified in vitro as a bioactive component present in FAHF‐2 and B‐FAHF‐2. The aim of this study was to investigate berberine as a chemical and pharmacokinetic marker of B‐FAHF‐2. Methods: The consistency of constituents between B‐FAHF‐2 and FAHF‐2 was tested. Peanut allergic C3H/HeJ mice were orally administered with 1 mg of berberine or B‐FAHF‐2 containing an equivalent amount of berberine, and the ability to protect against peanut anaphylaxis and pharmacokinetic parameters were determined. Human intestinal epithelial cells (Caco‐2) were cultured with berberine with or without the nine individual herbal constituents in B‐FAHF‐2, and the absorbed berberine levels were determined. Results: Berberine is one of the major components in B‐FAHF‐2 and FAHF‐2 formula. In a peanut allergic mouse model, B‐FAHF‐2, but not berberine, protected mice from anaphylaxis reactions. Pharmacokinetic profiles showed that the Cmax of B‐FAHF‐2 fed mice was 289.30 ± 185.40 ng/mL; whereas berberine alone showed very low bioavailability with Cmax value of 35.13 ± 47.90 ng/mL. Caco‐2 cells influx assay showed that 7 of 9 herbal constituents in B‐FAHF‐2 increased berberine absorption at rates ranging from 18 to 205%. Conclusions: B‐FAHF‐2 remarkably increased the bioavailability of berberine. Berberine can be used as chemical and pharmacokinetic marker of B‐FAHF‐2. Other herbal components in B‐FAHF‐2 may facilitate the absorption of berberine. HighlightsBerberine is the major component in both FAHF‐2 and B‐FAHF‐2 formula.Berberine alone did not protect peanut‐allergic mice from anaphylaxis reaction in vivo, while the B‐FAHF‐2 formula did.Berberine showed very poor bioavailability in mice, while the B‐FAHF‐2 formula increased the berberine absorption rate.7 out of 9 herbal constituents in B‐FAHF‐2 formula showed enhanced absorption of berberine by Caco‐2 cells in vitro.

The Flavonoid 7,4′-Dihydroxyflavone Prevents Dexamethasone Paradoxical Adverse Effect on Eotaxin Production by Human Fibroblasts

Eotaxin/CCL‐11 is a major chemoattractant that contributes to eosinophilic inflammation in asthma. Glucocorticoids inhibit inflammation, but long‐time exposure may cause paradoxical adverse effects by augmenting eotaxin/CCL‐11production. The aim of this study was to determine if 7,4′‐dihydroxyflavone (7,4′‐DHF), the eotaxin/CCL11 inhibitor isolated from Glycyrrhiza uralensis, reduces in vitro eotaxin production induced by long‐time dexamethasone (Dex) exposure, and if so, to elucidate the mechanisms of this inhibition. Human lung fibroblast‐1 cells were used to identify the potency of 7,4′‐DHF compared with other compounds from G. uralensis, to compare 7,4′‐DHF with Dex on eotaxin production following 24‐h short‐time culture and 72‐h longer‐time (LT) culture, and to determine the effects of the 7,4′‐DHF on Dex LT culture augmented eotaxin production and molecule mechanisms. 7,4′‐DHF was the most potent eotaxin/CCL‐11 inhibitor among the ten compounds and provided continued suppression. In contrast to short‐time culture, Dex LT culture increased constitutively, and IL‐4/TNF‐α stimulated eotaxin/CCL11 production by human lung fibroblast‐1 cells. This adverse effect was abrogated by 7,4′‐DHF co‐culture. 7,4′‐DHF significantly inhibited Dex LT culture augmentation of p‐STAT6 and impaired HDAC2 expression. This study demonstrated that 7,4′‐DHF has the ability to consistently suppress eotaxin production and prevent Dex‐paradoxical adverse effects on eotaxin production. Copyright