Background: Populus tremula L. (Poplar), Fraxinus excelsior L. (ash) and Solidago virgaurea L. (goldenrod) have been used for medicinal purposes through centuries, to treat pain, fever and inflammation, but their mechanisms of action are still not fully understood. The present study was performed to investigate, whether the herbal medicinal product Phytodolor (R) (STW 1) and its components have anti-inflammatory effects on activated human monocytes and differentiated human macrophages to elucidate their modes of action in comparison with well-known analgesic, non-steroidal anti-inflammatory drug (NSAIDs) as diclofenac. Methods: Adherent human monocytes obtained from peripheral blood mononuclear cells (PBMCs) were cultured in serum-free medium and pre-treated with 50-100 mu g/ml of diclofenac, STW 1, their components, poplar, ash or goldenrod or its combination (0.05% to 2%). Thereafter, monocytes were activated with 0.1 or 1 mu g/ml LPS for 24 h. The intracellular expressions of TNF-alpha or PTGS2 were determined by cell-based ELISA. Apoptotic cells were identified by YO-PRO-1 staining. Protein or total RNA were isolated to perform SDS-PAGE/Western blot and qRT-PCR analyses. PMA-differentiated human THP-1 macrophages were pre-treated with diclofenac (50 mu g/ml) or STW1 (0.1%) and afterwards with LPS (1 mu g/ml) and the translocation of the intracellular p62 NF-kappa B subunit was detected by immunofluorescence. Results: STW 1 inhibited the intracellular content of TNF-alpha and PTGS2 protein, as well as of TNF-alpha and PTGS2 gene expression and induced apoptosis in LPS-activated human monocytes under serum free conditions. Furthermore, STW 1 inhibited the translocation of the p65 subunit of the redox-regulated NF-kappa B into the nucleus in LPS-activated human macrophages. Conclusion: The present in vitro investigations suggest a significant anti-inflammatory activity of STW 1 and its components by inhibiting pro-inflammatory cytokine as TNF-alpha and the key enzyme PTGS2 in LPS-activated human monocytes, which is, at least partly mediated through the suppression of NF-kappa B activation. Our results provide evidence for distinctive anti-inflammatory effects of STW 1 and its components on LPS-activated human monocytes/macrophages and, thus, for the therapeutic use of STW 1 in inflammation and pain related disorders.

While the use of St John's wort extracts as treatment for mild to moderate depression is well established the mode of action is still under investiation. Individual constituents of St John's wort extract were tested for possible effects on the β1AR density and a subsequent change in downstream signalling in rat C6 glioblastoma cells.

or absence of pleural effusion or ascites did not differ between normal and delayed MTX elimination groups.Co-administration of PPI was more prevalent in delayed elimination group compared with normal elimination group (90% vs. 62%).No difference was found in NSAIDs or vancomycin co-administration between the groups.Multivariate logistic regression model adjusted for age and gender revealed that co-administration of PPI was an independent factor associated with delayed MTX elimination (OR 12.3, p=0.04).Conclusion: Co-administration of proton pump inhibitors delays elimination of plasma methotrexate in patients with high-dose methotrexate therapy.

PURPOSE:An herbal preparation, STW 5, used clinically in functional dyspepsia and irritable bowel syndrome, has been shown to possess properties that may render it useful in inflammatory bowel disease (IBD). The present work was conducted to study its effectiveness in a rat model of IBD.METHODS:An experimental model reflecting ulcerative colitis in man was adopted, whereby colitis was induced in Wistar rats by feeding them 5 % dextran sulfate sodium (DSS) in drinking water for one week. STW 5 and sulfasalazine (as a reference standard) were administered orally daily for 1 week before colitis induction and continued during DSS feeding. The animals were then sacrificed, and the severity of colitis was evaluated macroscopically and microscopically. Colon samples were homogenized for determination of reduced glutathione, tumor necrosis factor-α, and cytokine-induced neutrophil chemoattractant-3 as well as myeloperoxidase, glutathione peroxidase, and superoxide dismutase. In addition, colon segments were suspended in an organ bath to test their reactivity towards carbachol, KCl, and trypsin.RESULTS:STW 5 and sulfasalazine were both effective in preventing the shortening of colon length and the increase in both colon mass index and total histology score as well as the changes in biochemical parameters measured except changes in dismutase activity. DSS-induced colitis led to marked depression in colonic responsiveness to the agents tested ex vivo, an effect which was normalized by both drugs.CONCLUSIONS:The findings point to a potential usefulness of STW 5 in the clinical setting of ulcerative colitis.

Proton pump inhibitor (PPI) therapy is the most effective medical treatment for symptom relief in gastro-esophageal reflux disease (GERD). However, up to 40% of patients do not achieve adequate symptom relief, especially those suffering from non-erosive reflux disease (NERD) which does not respond well to PPIs. Therefore, the search continues for other treatment options. STW 5, a multi-component herbal preparation, was shown to relieve heartburn and concomitant reflux symptoms in patients with functional dyspepsia and to prevent inflammation in an acute model of reflux esophagitis (RE), without affecting the pH of the refluxate. The present study assesses the efficacy of STW 5 in a more chronic model of RE, and the underlying mechanisms both In-Vivo and in-vitro. Rats were pretreated for 7 d with STW 5 or omeprazole (as reference drug) before surgical induction of esophagitis. RE was achieved by ligation of the fore-stomach and covering the duodenum near the pylorus with a piece of Nelaton catheter. After recovery, rats were treated for a further 10 d with the drugs and then sacrificed, their esophagi excised, weighed and evaluated macroscopically. Tissue homogenates were used for semi-quantitative determination of cytokines using a cytokine array. Both, STW 5 and omeprazole, improved body weight, tissue damage score and esophageal weight index to similar extents. However, STW 5 had a much more pronounced anti-inflammatory effect. As shown in the cytokine array (fig. 1), STW 5 inhibited the majority of the measured pro-inflammatory cytokines induced by surgical reflux, suggesting a direct anti-inflammatory and/or mucosal protecting action. Since recent evidence indicates that the mucosal damage observed in GERD is due to release of inflammatory mediators from mucosal and sub-mucosal cells in response to bile salts and other substances in the gastric refluxate, the effects of STW 5 on cytokine release from the normal human esophageal cell-line HET-1A in response to stimulation with chenodeoxycholic acid (CDCA) was assessed. Incubation of cells with CDCA caused marked release of CD40ligand, IFN-γ, IL-1ra, IL-6, IL-8 and IL-23, which were all inhibited by co-incubation with STW 5 (0,3 10 μl/ml) as assessed semi-quantitatively (cytokine array, fig.2) and confirmed quantitatively (ELISA), without affecting cell viability. Similar effects were observed when cells were incubated with capsaicin as TRPV1 agonist (shown to be the main receptor involved in acid induced cytokine release from HET-1A cells), which evoked the release of CD40-ligand and IL-23. The present findings suggest that multi-target anti-inflammatory drugs like STW 5 might present an alternative/additional treatment option for GERD patients not responding adequately to PPIs. Further elucidation of its exact mechanism of action might pave the way for a new class of anti-reflux agents.

Background and Aims: The question was raised, whether the herbal medicine STW 5 acts on contractions elicited after electrical field stimulation (EFS) and on intestinal slow wave activity in the small intestine of mice, in a new type of an In Vitro model of trinitrobenzene sulfonic acid (TNBS) triggered inflammation.Methods: In the organ bath, segments of distal ileum of Balb/C mice were used for registration of spontaneous and EFS stimulated contractions.In the electrophysiological studies, segments of distal ileum (with minimum 50% increase in EFS induced contraction after 90 min of intraluminal TNBS application) were used for intracellular recordings, after removal of mucosa and submucosa.STW 5 was tested in a dilution of 1:100 in an organ bath and compared to vehicle (31% ethanol solution diluted 1:100), following intraluminal application of TNBS (0,01M solution, 3 cm H2O pressure) or Krebs solution for 90 min.Results: TNBS induced a significant time dependent enhancement of contractility (90 min: 155.1 ± 6.5% vs. control, n=15).STW 5 applied into the organ bath reduced basal tone (-22.1±2.1% TNBS vs. -17.7±1.5% no TNBS n=15), EFS induced contractility (-61.3±3.5% TNBS vs. -67.6±3.7% no TNBS n=10) and significantly prevented TNBS induced increase of EFS induced contractions (97.4 ± 4.1% STW 5+TNBS vs. TNBS/no STW 5 155.1 ± 6.5% n=10).Electrophysiological parameters were not significantly influenced in TNBS pretreated or control preparations.No significant effects on intracellular recordings of resting membrane potential, slow wave amplitude and frequency were observed making neuronal and muscular effects unlikely.Summary and Conclusions: Stimulation with TNBS results in inflammation induced changes of motility, which are reversed or prevented by STW 5.The underlying mechanisms may be of relevance in the treatment of functional gastrointestinal and subclinical inflammatory gastrointestinal diseases.

Das Phytopharmakon STW 5 (Iberogast®) besteht aus neun Pflanzenextrakten und wird erfolgreich zur Therapie des Reizdarmsyndroms eingesetzt. Während die Wirkung von STW 5 gut belegt ist, sind zum Beitrag der enthaltenen Pflanzenextrakte zur Wirkung des Gesamtextraktes bisher noch Fragen offen. Ziel unserer Studie war es, den Einfluss der Extraktkombination STW 5-II und einiger ihrer Komponenten auf die Acetylcholin (ACh)-induzierte Kontraktion an intakten und entzündlich veränderten Dünndarmpräparaten der Ratte im Vergleich zu STW 5 zu untersuchen.

Irritable bowel syndrome (IBS) is a disease in which, typically, alterations in intestinal motility and visceral hypersensitivity appear to exist, apparently without any organic alteration (Thompson, 1991). Several pathogenic factors responsible for IBS have been suggested. It seems that there are cell factors, which give reason to believe that there is a low-grade intestinal inflammation in this pathology (Ortiz-Lucas et al., 2010). Several cytokines, such as tumour necrosis factor α (TNFα), interleukin 1, and interleukin 6, contribute to the pathogenesis (Ardizzone and Bianchi Porro, 2005; Pizarro et al., 2006). Macrophages are the major producers of TNFα, and, interestingly, they are also highly responsive to TNFα. TNFα has been shown to play a pivotal role in activating the cytokine cascade in many inflammatory diseases and it has been proposed as a therapeutic target for a number of diseases. Consequently, recent strategies for the treatment of intestinal inflammation have primarily targeted the immunopathogenic processes that mediate intestinal inflammation at the cytokine level (Bamias et al., 2003; Sandborm and Targan, 2002). At present, pharmacotherapy represents the mainstay of inflammatory bowel disease management (Stein and Hanauer, 1999). Some anti-inflammatory or immuno-modulating drugs, including salicylates and methotrexate, are able to decrease intracellular adenosine 5 -triphosphate concentrations and raise extracellular adenosine levels. It has been proposed that such properties can significantly contribute to the drugs’ pharmacological actions in inflammatory diseases (Cronstein et al., 1999). Several lines of evidence suggest that adenosine regulates immunity and inflammation (Amann and Peskar, 2002; Montesinos et al., 2007). The wide distribution of adenosine receptors (AR) as well as enzymes for purine metabolism in different gut regions suggests a complex role for this mediator in the regulation of gastrointestinal functions (Antonioli et al., 2008). Adenosine binds to four different types of G protein-coupled cell surface receptors referred to as A1R, A2AR, A2BR, and A3R, each having a unique pharmacological profile, tissue distribution and signalling pathway (Jacobson and Gao, 2006). All known ARs

STW 5 (Iberogast®), an established herbal combination, was effective in randomized, double blind clinical studies in functional dyspepsia and irritable bowel syndrome. Since STW 5 was found to influence intestinal motility and has anti-inflammatory properties, this study investigated the expression of adenosine receptors and characterized their role in the control of the anti-inflammatory action of STW 5 and its fresh plant component STW 6 in inflammation-disturbed rat small intestinal preparations. The inflammation was induced by intraluminal instillation of 2,4,6-trinitrobenzene sulfonic acid (TNBS, 0.01 M). The effects of coincubation with selective receptor agonists and antagonists, STW 5, STW 6, or combinations of these compounds on acetylcholine (ACh)-evoked contraction of ileum/jejunum preparations were tested. Adenosine receptor mRNA expression was examined by reverse transcription-polymerase chain reaction (RT-PCR). In untreated preparations, RT-PCR revealed the presence of all adenosine receptor subtypes. Suppressed expression was detected for all subtypes in inflamed tissues, except for A(2B)R mRNA, which was unaffected. STW 5 reversed these effects and enhanced A(2A)R expression above control levels. Radioligand binding assays confirm the affinity of STW 5 to the A(2A)R, and the A(2A)R antagonist was able to prevent the effect of STW 5 on TNBS-induced attenuation of the ACh contraction. Our findings provide evidence that STW 5, but not STW 6 interacts with A(2A)R, which is involved in the anti-inflammatory action of STW 5. STW 6 did not contribute to adenosine A(2A)R-mediated anti-inflammatory effect of STW 5. Other signaling pathways could be involved in the mechanism of action of STW 6.

Summary All processes of oxygen activation include very reactive intermediates. Therefore, aerobic cells must cope with - and to some extent also adapt to - oxidative stress provoked for example by infections or intoxications, where these reactive intermediates accumulate. All inflammatory processes include such oxygen activating processes where reactive oxygen species (ROS) are produced. Dependent on the strength of these impact(s), several symptoms indicate the deviation from normal, steady-state metabolism. Intrinsic radical scavenging processes or compounds administered with food thus have to warrant metabolic control within certain limits. Antioxidants which in many cases are free radical scavengers or quenchers of activated states comprise a vast number of classes of organic molecules including phenolics as the most prominent ones. In this publication the activities of extracts from Fraxinus excelsior, Populus tremula and Solidago virgaurea as components of the drug Phytodolor® and their mechanisms of protection from oxidative damage are summarized. In addition, new results on tyrosine nitration, a process characteristic for sites of inflammation, and its inhibition by these plant extracts, is reported.

To examine mechanisms underlying the role of chronic stress in post-infectious irritable bowel syndrome (PI-IBS), we previously studied the effect of chronic stress and prior infectious colitis in the C. rodentium infected mouse (day 30 post infection), a model of human E. coli self-limiting colitis (NGM A:257,2010).Water avoidance stress (WAS; 1 hr on days 21-30 following infection) increased stress hormones (corticosterone and epinephrine), excitability of colonic nociceptive DRG neurons and colonic multi-unit afferent firing, compared to post-infected animals alone The current study examined pathways underlying this stress-post-infection effect using patch clamp recordings from Fast Blue labeled colonic DRG neurons to measure changes in excitability (rheobase (Rh) and/or increases in action potential discharge (APD)).At day 30, infection had resolved and histopathology was normal.However, tissue trypsin-like activity (>10 fold; p<0.01) and serine and cysteine proteases (~0.5-1 fold; p < 0.05) were elevated in unstressed post-infected animals.In patch clamp studies, excitability of neurons incubated in colonic tissue supernatants from unstressed post-infected mice was significantly inhibited by a global protease inhibitor (post-infected Rh = 61.5 +/-7.5 pA vs. post-infected + inhibitor = 96.3+/-11.5 pA; p = 0.0162 and post-infected APD = 4.4 +/-0.7 vs. post-infected + inhibitor 1.8 +/-0.4;p = 0.007).In contrast, neuronal excitability with supernatants from uninfected WAS animals was not altered by the protease inhibitor and excitability did not differ between neurons exposed to WAS or control supernatants.Colonic histopathological scoring and Western blots of tight junction proteins (occludin) were also not different between WAS and control groups.We therefore tested whether stress hormones could signal directly to DRG neurons.Labeled neurons were isolated using laser captured microdissection and corticosterone and β2 receptor mRNA identified by PCR.In patch clamp studies, colonic DRG neurons incubated in epinephrine (5 nM) and corticosterone (1 μM) were hyperexcitable compared to controls (epinephrine /corticosterone Rh = 48.6 +/-10.9pA vs control = 85.8 +/-12.3pA, p = 0.03 and epinephrine/corticosterone APD = 2.6 +/-0.4 vs. control = 1.6 +/-0.2;p = 0.04).Compared to controls, incubating neurons with protease activating receptor2 -activating peptide (PAR2-AP; 30 μM) or WAS alone had no effect on Rh.However, WAS and PAR2-AP (30 μM) combined markedly decreased Rh (control + PAR2-AP = 85.8 +/-12.3pA vs. WAS + PAR2-AP 19.2 +/-3.5 pA, p = 0.002).These data suggest tissue proteases and circulating stress hormones converge on DRG nociceptive neurons to increase sensory signaling from the colon.Moreover, this interaction enables sub-threshold levels of proteases to enhance peripheral sensory signaling.

Abstract The well-known anti-inflammatory and analgesic effects of the phytopharmacon willow bark extract have been attributed to the content of salicin; however, pharmacological studies have shown that salicin alone, despite being involved in its therapeutic action, cannot fully explain its clinical efficacy. In addition to reducing inflammation and pain, acetylsalicylic acid (ASA, CAS 50-78-2), like other synthetic non-steroidal anti-inflammatory drugs (NSAIDs), has been shown to exert anti-proliferative effects and to induce apoptosis in a variety of cell lines, e. g. colon, stomach, and prostate cancer cells. To investigate the mechanism of action and possible anti-proliferative and pro-apoptotic effects of willow bark, a water extract (STW 33-1) and a polyphenol rich fraction (fraction E) have been tested by using the colon-carcinoma cell line HT-29. Both, STW 33-1 and its fraction E showed significant anti-proliferative and pro-apoptotic effects on HT-29 cancer cells. Related to the salicin content of the willow bark extract, a higher dosage of ASA was needed. Furthermore, compared to ASA and to diclofenac (Diclo, CAS 15307-79-6), the COX-1 and COX-2 mRNA expressions were influenced differently by STW 33-1 and fraction E. ASA and Diclo inhibited both the COX-1 and COX-2 mRNA expressions, whereas STW 33-1 and its fraction E increased the COX-1 mRNA expression. In addition to the already well-known anti-inflammatory and analgesic effects, willow bark extract has been found to possess anti-proliferative and pro-apoptotic effects similar to NSAIDs. The different influence of willow bark on the COX-1 and COX-2 mRNA expressions in comparison to NSAIDs might be relevant, e. g. for prevention of undesirable side effects such as gastric erosions.

G A A b st ra ct s patients were re-assigned to LL (n=84) or LP (n=86) group. Mean changes from baseline at all treatment weeks were statistically significant in all treatment groups for stool consistency, abdominal discomfort/pain, and SBM frequency (p<0.003). L group had statistically significant improvement in stool consistency compared to P group in almost a half of the 12 weekly visits (p<0.03). Similar significant improvements were observed at Weeks 13, 14, and 15 for the LL group (p<0.05). L group had statistically significant mean reduction from baseline in abdominal discomfort/pain at Weeks 10 and 11 compared to P group (p<0.05). Significant results were further observed at Weeks 13, 15, and 16 for the LL group (p<0.04). These results were not seen in the LP group. The range of increase from baseline in SBM frequency at Period I was 2.1-2.9 and 1.4-2.5 for the L and P groups, respectively. LL group had a range of 2.6-3.2, whereas the range for LP group and P group were 2.0-2.7 and 2.12.5, respectively. CONCLUSION: Lubiprostone provided significant improvement in stool consistency, abdominal discomfort/pain, and SBM frequency for standard treatment of 12 weeks, and consistent results of all outcomes were observed during 16 weeks of treatment. Moreover, longer term treatment provided significant reduction in abdominal discomfort/ pain.

Summary Extracts from the herb “St. John’s wort” (Hypericum perforatum L.) exhibit beneficial effects on patients suffering from mental depressions. Lack of catecholamine neurotransmitters may be one biochemical mechanism for this problem under discussion. It has been recently reported that alcoholic extracts from Hypericum perforatum inhibit dopamine-ß-hydroxylase (D-ß-H) with an I50 of 0.1 µmol/1 on the basis of total hypericin content and with an I50 of 21 µmol/1 with pure commercial hypericin. As test system polarographic determination of oxygen uptake with tyramine as a substrate analogue was used. In the present paper the quantification of the enzymatic activity and the potential influence of inhibitors are reported using dopamine as substrate and product (noradrenaline) quantification by HPLC. With this test system it could be shown that D-ß-H is strongly inhibited by pseudohypericin (I50 = approx. 3 µmol/1) and hypericin (I50 = approx. 5 µ.mol/1), whereas the I50-values of various flavonoids (quercitrin, isoquercitrin, hyperoside, rutin, quercetin, amentoflavone, kaempferol) are in the range of 50 µ.mol/1 or higher.

Experimental study on plaque progression, regression and composition in atherosclerotic thoracic aorta of hypercholesterolemic rabbits after long-term withdrawal of cholesterol-enriched diet (CED).Rabbits were fed 2% cholesterol for 6 weeks followed by withdrawal periods for 15, 23, 34, 68, or 78 weeks. Cholesterol, triglyceride, and phospholipids levels in blood and cholesterol concentrations in aorta were quantified. Plaque size and cellularity, phenotype of macrophages and smooth muscle cells were (immuno)histomorphometrically analyzed in segments of the thoracic aorta.After 6 weeks of CED, blood cholesterol levels were about 80-fold higher, whereas atherosclerosis and cholesterol content in the thoracic aorta were only minimally increased. However, the latter significantly increased within 15 weeks after cholesterol withdrawal, while serum cholesterol level was still 10-fold increased. Thereafter plaque area and cholesterol content remained almost unchanged until the end of the study despite a long-term normalization of serum cholesterol level after withdrawal of CED. Directly after 6 weeks of CED the densities of macrophages and apoptotic cells within plaques were highest, decreasing after cholesterol withdrawal, whereas, vice versa the density of smooth muscle cells (SMCs) significantly increased.We suggest that atherosclerotic plaques respond to long-term withdrawal of CED by decrease in number and phenotype of macrophages and increase of SMCs without regression of the lesion size. The cellular changes are suggested to considerably contribute to higher plaque stability.

A multitarget herbal preparation, STW 5, has been used clinically in different gastrointestinal disorders including functional dyspepsia and irritable bowel syndrome. Previous studies have shown that it possesses properties that may render it useful in gastro-oesophageal reflux disease (GERD). We performed this study to test this compound in an acute model of reflux oesophagitis in rats. Oesophagitis was induced surgically by ligating the pyloric end and fore-stomach. Lower oesophageal pH was measured 3 h later in conscious animals. Five hours after surgery, animals were sacrificed and the oesophagi were examined macroscopically and histologically. Selected markers of inflammation were measured in oesophageal homogenates. STW 5 was given orally for 5 days before induction of oesophagitis. Pantoprazole was used as a reference standard. Ligated animals showed a high incidence of ulcerative lesions associated with a marked increase in myeloperoxidase, thiobarbituric acid reactive substances, tumor necrosis factor-alpha, and interleukin-1 beta. STW 5 did not affect oesophageal pH, but dose-dependently reduced the severity of the oesophageal lesions and normalized the deranged level of the inflammation markers. The beneficial effects were confirmed histopathologically. STW 5 proved to be effective in protecting against inflammatory lesions in this model of oesophagitis, thus warranting further investigation of its potential therapeutic usefulness in GERD.

Retrolective study of the long-term treatment of degenerative diseases of the locomotor system with STW 1 in a general practice

Das Phytotherapeutikum STW 5 (Iberogast®) wird seit fast 50 Jahren zur Therapie von Magen-Darm-Erkrankungen eingesetzt. Es ist zugelassen zur Behandlung funktioneller Magen-Darm-Erkrankungen wie funktionelle Dyspepsie und Reizdarmsyndrom. Für diese Indikationen liegen umfangreiche wissenschaftliche Daten in Form von placebokontrollierten, randomisierten Doppelblindstudien, Metaanalysen, Anwendungsbeobachtungen, retrospektiven Erhebungen sowie Subgruppenanalysen vor.