In vitro studies on the mechanisms of action of chamomile, myrrh and coffee charcoal – components of a traditional herbal medicinal product (Myrrhinil-Intest®)

The traditional herbal medicinal product Myrrhinil-Intest® is a fixed herbal combination, which is marketed in Germany since 1959 and applied in medical practice for the treatment of gastrointestinal disorders such as functional diarrhea, irritable bowel syndrome and inflammatory bowel disease. It contains myrrh, which is described as the oleo-gum resin from mainly Commiphora molmol Engler (Burseraceae), coffee charcoal, which are the milled roasted to blackening outer seed parts of green dried Coffea Arabica Linné (Rubiaceae) fruits and chamomile flowers - the flower heads of Matricaria recutita Linné (Asteraceae). The clinical effectiveness of Myrrhinil-Intest® for the treatment of various gastrointestinal disorders was demonstrated in several clinical studies and is described in various experience reports, however its pharmacological profile is not fully elucidated. Within the present study the spasmolytic and anti-inflammatory potential of the components myrrh, chamomile and coffee charcoal was investigated. Therefore pharmacological, histological and molecular biological methods were utilised. Spasmolytic activity was characterised using isometric tension measurement with rat isolated small intestinal preparations. Anti-inflammatory potential was assessed with different methods using isolated rat small intestinal preparations and immune cell lines. Inflammation was induced with TNBS and LPS respectively. Additionally, the influence of the herbal components on the gene expression profile of native human macrophages after LPS/IFNγ stimulation was determined by microarray gene expression analysis. Chamomile flower and myrrh exerted spasmolytic effects, whereby the more pronounced spasmolytic effects of myrrh were mediated via calcium channel blockade. Myrrh and chamomile flower exerted anti-inflammatory effects

Antifungal Activities Of Essential Oils Of Cymbopogon Citratus, Ocimum Gratissimum And Eucalyptus Globulus Against Botrytis Cinerea, Penicillium Expansum And Potential Application As Biocontrol Agents During Fruit Cultivation

The present work aims to study the effectiveness of selected medicinal and food plant extracts (EtOH, DCM) including essential oils (Eucalyptus globulus, Ocimum gratissimum, Cymbopogon citractus) on the mycellian growth of toxinogenic moulds responsible for fruit alteration. Antifungal tests were carried out with the different extracts to assess their potential antimicrobial activities against Botrytis cinerea and Penicillium expansum responsible for fruits deterioration in several regions of the world.In vitro antifungal tests have shown that essential oils have pronounced antifungal activities at low concentrations of 0.3µl/ml, 1.3µl/ml and 5µl/ml respectively for Cymbopogon citractus, Eucalyptus glogulus and Occimum gratissimum, while non-volatile extracts exert growth inhibition of 50% on average on Botrytis cinerea. This study suggests use as biocontrol agents for plant pests during fruit cultivation that plants present themselves as a promising alternative for the fight against toxinogenic fungi

Bioactive Plant Compounds in Coffee Charcoal (Coffeae carbo) Extract Inhibit Cytokine Release from Activated Human THP-1 Macrophages

The herbal preparation coffee charcoal is produced by over-roasting and milling green dried Coffea arabica L. seeds, and has a long-standing tradition in the treatment of inflammatory and gastrointestinal disorders. Its therapeutic properties are commonly attributed to adsorptive and astringent effects. This insufficiently explains its mode of action, especially when used in the treatment of inflammatory diseases in lower dosages. Our investigations aimed to identify bioactive secondary plant metabolites affecting cytokine-signaling. Thus, a phytochemical analysis of coffee charcoal extract was conducted using HPLC and LC/MS. Trigonelline, neochlorogenic acid, chlorogenic acid, caffeine, cryptochlorogenic acid, feruloylquinic acid isomers, and a caffeoylquinolacton were identified in the extract. Subsequently, the effects of coffee charcoal extract, chlorogenic acid isomers, their metabolite caffeic acid, caffeine, and trigonelline on cytokine (TNF, IL-6, MCP-1) release from LPS-challenged human THP-1 macrophages were examined to evaluate anti-inflammatory activity. Coffee charcoal showed concentration-dependent mild-to-medium inhibitory effects. The chlorogenic acid isomers and caffeic acid inhibited the TNF release, with cryptochlorogenic acid exerting the most distinct effects, as well as decreasing the release of IL-6 and MCP-1. In addition, scanning electron microscopic images provided an impression of the particle constitution, indicating a larger particle size and less structured surface of coffee charcoal in comparison to activated charcoal. In conclusion, our findings underline that beyond adsorptive effects, coffee charcoal exhibits pharmacological properties, which derive from a spectrum of secondary plant metabolites and support the therapeutic use in inflammatory diseases. Chlorogenic acids, particularly cryptochlorogenic acid, appear as pivotal bioactive compounds

Anti-Inflammatory Potential of Phenolic Compounds Isolated From Entada africana Guill. & Perr. Used in the Republic of Benin

In West African medicine, Entada africana Guill. & Perr. from the family of Fabaceae is used to treat inflammatory conditions in the management of fractures, wounds, and sprains in the northern region of the Republic of Benin. The aim of the present study was to isolate and elucidate phenolic compounds from a hydroalcoholic leaf extract from E. africana and to identify compounds with anti-inflammatory activity in vitro. Eleven compounds were purified from three fractions, which have shown strong to medium anti-inflammatory activity. The isolated compounds were characterized by HRESI-MS and NMR methods as gallic acid (1), ethyl gallate (2), 5,7-dihydroxychromen-4-one (3), 3′,4′,7-trihydroxyflavone (4), dihydrokaempferol-7-O-glucoside (5), catechin (6), quercetin-3-O-[β-apiosyl- (1‴→2″)-β-glucoside] (7), quercetin-3-O-glucoside (8), naringenin-7-O-glucoside (9), aromadendrin (10), and myricetin-3-O-glucoside (11). Nine of the major phenolic compounds were tested using TNF-α stimulated human keratinocytes (HaCaT) as skin inflammation model to identify molecules, which may explain the use of the plant leaves as an anti-inflammatory remedy by assessing the release of proinflammatory cytokines IL-8 and IL-6. The hydroacoholic leaf extract of E. africana exerted a medium inhibitory effect on the release of IL-8. 3′,4′,7-trihydroxyflavone, aromadendrin, dihydrokaempferol-7- O-glucoside and ethyl gallate demonstrated a strong to medium effect on the release of IL-6. For the release of IL-8, 3′,4′,7-trihydroxyflavone demonstrated a medium activity. This study provides for the first time a detailed screening of phenolic compounds occurring in the hydroethanolic leaf extract of E. africana. Additionally, it is shown that E. africana contains active compounds which may justify its traditional medicinal use as an antiinflammatory remedy to treat inflammatory and pain-related skin conditions in the Republic of Benin

Phytochemical Characterization and In Vitro Anti-Inflammatory, Antioxidant and Antimicrobial Activity of Combretum Collinum Fresen Leaves Extracts from Benin

Leaves from Combretum collinum Fresen (Combretaceae) are commonly used for the treatment of inflammatory conditions, wound healing and bacterial infections in traditional West African medicine. This research focuses on the characterization of the phenolic profile and lipophilic compounds of leaves extracts of C. collinum. Studies of the in vitro anti-inflammatory activity were performed in TNFα stimulated HaCaT cells and antibacterial activity was evaluated with agar well diffusion and microdilution assays. Antioxidant activity was determined by DPPH and ABTS assays and compared to standards. The phytochemical studies confirmed myricetin-3-O-rhamnoside and myricetin-3-O-glucoside as major components of the leaves extracts, each contributing significantly to the antioxidant activity of the hydrophilic extracts. GC-MS analysis identified 19 substances that were confirmed by comparison with spectral library data and authentic standards. Combretum collinum aqueous leaves extract decreased pro-inflammatory mediators in TNFα stimulated HaCaT cells. Further investigations showed that myricetin-3-O-rhamnoside has an anti-inflammatory effect on IL-8 secretion. In the antimicrobial screening, the largest inhibition zones were found against S. epidermidis, MRSA and S. aureus. MIC values resulted in 275.0 µg/mL for S. epidermidis and 385.5 µg/mL for MRSA. The in vitro anti-inflammatory, antibacterial and antioxidant activity supports topical use of C. collinum leaves extracts in traditional West African medicine

Chemical Profile and Antimicrobial Activity of the Fungus-Growing Termite Strain Macrotermes Bellicosus Used in Traditional Medicine in the Republic of Benin

The fungus growing termite species Macrotermes bellicosus (M. bellicosus) is used in nutrition and traditional medicine in the Republic of Benin for the treatment of infectious and inflammatory diseases. Previous findings demonstrated evidence of anti-inflammatory and spasmolytic properties of M. bellicosus. The aim of the present study was to evaluate the antimicrobial potential of different extracts of M. bellicosus samples and determine the chemical profile of an ethanolic M. bellicosus extract. Chemical profiling was conducted using centrifugal partition chromatography and 13C-NMR, followed by MALDI-TOF MS. Major identified compounds include hydroquinone (HQ), methylhydroquinone (MHQ), 3,4-dihydroxyphenethyl glycol (DHPG), N-acetyldopamine (NADA) and niacinamide. The fatty acid mixture of the extract was mainly composed of linoleic and oleic acid and highlights the nutritional purpose of M. bellicosus. Using the Kirby–Bauer disc diffusion and broth microdilution assay, an antibacterial activity of M. bellicosus samples was observed against various clinical strains with a highest growth inhibition of S. aureus. In addition, HQ and MHQ as well as fractions containing DHPG, niacinamide and NADA inhibited S. aureus growth. The reported antimicrobial activity of M. bellicosus and identified active substances provide a rationale for the traditional medicinal use of M. bellicosus

A focus on critical aspects of uptake and transport of milk-derived extracellular vesicles across the Caco-2 intestinal barrier model

Bovine milk-derived extracellular vesicles (EVs) hold promises as oral drug delivery systems. Since EV bioavailability studies are difficult to compare, key factors regarding EV uptake and intestinal permeability remain little understood. This work aims to critically study uptake and transport properties of milk-derived EVs across the intestinal barrier in vitro by standardization approaches. Therefore, uptake properties were directly compared to liposomes in intestinal Caco-2 cells. Reliable staining results were obtained by the choice of three distinct EV labeling sites, while non-specific dye transfer and excess dye removal were carefully controlled. A novel fluorescence correction factor was implemented to account for different labelling efficiencies. Both EV and liposome uptake occurred mainly energy dependent with the neonatal Fc receptor (FcRn) providing an exclusive active pathway for EVs. Confocal microscopy revealed higher internalization of EVs whereas liposomes rather remained attached to the cell surface. Internalization could be improved when changing the liposomal formulation to resemble the EV lipid composition. In a Caco-2/HT29-MTX co-culture liposomes and EVs showed partial mucus penetration. For transport studies across Caco-2 monolayers we further established a standardized protocol considering the distinct requirements for EVs. Especially insert pore sizes were systematically compared with 3 µm inserts found obligatory. Obtained apparent permeability coefficients (Papp) reflecting the transport rate will allow for better comparison of future bioavailability testing

In vitro studies on the mechanisms of action of chamomile, myrrh and coffee charcoal – components of a traditional herbal medicinal product (Myrrhinil-Intest®)

The traditional herbal medicinal product Myrrhinil-Intest® is a fixed herbal combination, which is marketed in Germany since 1959 and applied in medical practice for the treatment of gastrointestinal disorders such as functional diarrhea, irritable bowel syndrome and inflammatory bowel disease. It contains myrrh, which is described as the oleo-gum resin from mainly Commiphora molmol Engler (Burseraceae), coffee charcoal, which are the milled roasted to blackening outer seed parts of green dried Coffea Arabica Linné (Rubiaceae) fruits and chamomile flowers - the flower heads of Matricaria recutita Linné (Asteraceae). The clinical effectiveness of Myrrhinil-Intest® for the treatment of various gastrointestinal disorders was demonstrated in several clinical studies and is described in various experience reports, however its pharmacological profile is not fully elucidated. Within the present study the spasmolytic and anti-inflammatory potential of the components myrrh, chamomile and coffee charcoal was investigated. Therefore pharmacological, histological and molecular biological methods were utilised. Spasmolytic activity was characterised using isometric tension measurement with rat isolated small intestinal preparations. Anti-inflammatory potential was assessed with different methods using isolated rat small intestinal preparations and immune cell lines. Inflammation was induced with TNBS and LPS respectively. Additionally, the influence of the herbal components on the gene expression profile of native human macrophages after LPS/IFNγ stimulation was determined by microarray gene expression analysis. Chamomile flower and myrrh exerted spasmolytic effects, whereby the more pronounced spasmolytic effects of myrrh were mediated via calcium channel blockade. Myrrh and chamomile flower exerted anti-inflammatory effects

In vitro studies on the mechanisms of action of chamomile, myrrh and coffee charcoal – components of a traditional herbal medicinal product (Myrrhinil-Intest®)

The traditional herbal medicinal product Myrrhinil-Intest® is a fixed herbal combination, which is marketed in Germany since 1959 and applied in medical practice for the treatment of gastrointestinal disorders such as functional diarrhea, irritable bowel syndrome and inflammatory bowel disease. It contains myrrh, which is described as the oleo-gum resin from mainly Commiphora molmol Engler (Burseraceae), coffee charcoal, which are the milled roasted to blackening outer seed parts of green dried Coffea Arabica Linné (Rubiaceae) fruits and chamomile flowers - the flower heads of Matricaria recutita Linné (Asteraceae). The clinical effectiveness of Myrrhinil-Intest® for the treatment of various gastrointestinal disorders was demonstrated in several clinical studies and is described in various experience reports, however its pharmacological profile is not fully elucidated. Within the present study the spasmolytic and anti-inflammatory potential of the components myrrh, chamomile and coffee charcoal was investigated. Therefore pharmacological, histological and molecular biological methods were utilised. Spasmolytic activity was characterised using isometric tension measurement with rat isolated small intestinal preparations. Anti-inflammatory potential was assessed with different methods using isolated rat small intestinal preparations and immune cell lines. Inflammation was induced with TNBS and LPS respectively. Additionally, the influence of the herbal components on the gene expression profile of native human macrophages after LPS/IFNγ stimulation was determined by microarray gene expression analysis. Chamomile flower and myrrh exerted spasmolytic effects, whereby the more pronounced spasmolytic effects of myrrh were mediated via calcium channel blockade. Myrrh and chamomile flower exerted anti-inflammatory effects

Myrrh and Chamomile Flower Extract Inhibit Mediator Release from IgE-stimulated Mast-Cell-Like RBL-2H3 Cells

Recent clinical evidence supports the efficacy of a traditional medicinal product (TMP) containing a combination of myrrh (Commiphora myrrha (Nees) Engl.), coffee charcoal (Coffea arabica L.), and chamomile flower dry extract (Matricaria chamomilla L.) in the therapy of diarrhea and inflammatory bowel disease. Mast cells seem to play a key role in the symptom generation of irritable bowel syndrome (IBS). To evaluate the use of the TMP in IBS treatment, the effects of the herbal extracts on the release of mast-cell mediators from stimulated RBL-2H3 cells were investigated. Therefore, degranulation was induced by phorbol-12-myristate-13-acetate (PMA) and calcium ionophore A13187 (CI) or IgE stimulation, and the amounts of released β-hexosaminidase and histamine were quantified. The extracts showed no effect on the mediator release of PMA- and CI-stimulated RBL-2H3 cells. Myrrh and chamomile were able to reduce the β-hexosaminidase release of IgE-stimulated cells, while myrrh showed stronger inhibition of the mediator release than chamomile, which reduced only IgE-stimulated histamine release. Thus, these results indicate a mechanistic basis for the use of the herbal combination of myrrh, coffee charcoal, and chamomile flower extract in the symptom-oriented treatment of IBS patients with diarrheal symptoms