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Sulfasalazine is a potent and specific inhibitor of IKB-kinases IKKa and IKKb
April 2000
AGAA615
3160
3162
SULFASALAZINE IS A POTENT AND SPECIFIC INHIBITOR OF IKB-KINASES IKKA AND IKKB. Christoph Kurt Weber, Susanne Liptay, Thomas Wirth, Guido Adler, Roland Schmid.. Vniversit8t Ulm, Vim, Germany; Universitst, Vim, Germany; Univeritxt Vim, Ulm, Germany; Univ of Vim, Vim, Germany. Background & Aims: NF-KBlRel has been implicated in the pathogenesis of inflammatory bowel disease (IBD). Various drugs known to be effective for the treatment of IBD such as glucocorticoids and salicylates have been shown to interfere with NF-KBlRel signaling. The aim of this study was to define the molecular mechanism by which sulfasalazine inhibits NF-KB activation. Methods: The effects of sulfasalazine and its moieties on NF-KB signaling were evaluated using electromobility shift-, transfection- and immune complex kinase-assays. The direct effect of sulfasalazine on IKB kinase (IKK) activity was investigated using purified recombinant IKKaand IKKl3proteins. Results: NF-kBlRel activity induced by the overexpression of NF-KBinducing kinase (NIK), IKKa, IKKI3, or consitutively active IKKaand IKKl3mutants was inhibited in a dose dependent manner by sulfasalazine. Sulfasalazine inhibited TNFa, PMA and IL-I induced activation of endogenous IKKs in colonic epithelial and Jurkat T cells. Sulfasalazine did not affect other cell1ular kinases such as MAP-kinases JNK and p38. Sulfasalazine diminished the catalytic activity of purified IKKaand IKKl3in vitro. In contrast, the metabolic moieties of sulfasalazine, 5-aminosalicylic acid and sulfapyridine were not effective. The decrease in substrate phosphorylation by IKKaandl3is associated with a decrease of autophosphorylation of IKKs and can be antagonized by excess ATP. Conclusions: These data suggest that sulfasalazine is a competitive antagonist of ATP binding of IKKaand IKKI3. This suppression of NF-kB activation by inhibition of IKK activity may contribute to the well-known anti-inflammatory and immunosuppressive effects of sulfasalazine.
CENTRAL PROCESSING OF RECTAL PAIN IN IBS PATIENTS: AN FMRI STUDY. Bruno L. Bonaz, Emmanuel Papillon, Monica Baciu, Christophe Segebarth, Richard Bost, Jean-Francois Le Bas, Jacques Fournet, Dept of Gastroenterology, Grenoble, France; INSERM V438, Grenoble, France. Patients with irritable bowel syndrome (IBS) have a chronic visceral hyperalgesia as represented by hypersensitivity to balloon distension (I). Numerous arguments favour a role of the central nervous system in this hypersensitivity (2). Functional imaging techniques of the human brain, such as fMRI, present considerable interest, as they permit to depict local changes in brain activation during visceral sensitive or nociceptive stimulation. In healthy volunteers, using fMRI, we recently characterize cerebral loci activated by a rectal distension as for example the insular cortex, anterior cingulate cortex (ACC) and dorso-Iateral prefrontal cortex (DLPFC) (3). Aim: to characterize, using fMRI, cerebral loci activated by a rectal distension in IBS patients. Methods: 8 right-handed patients (7 women, I man, median: 42 years) were consecutively recruited. Prior to fMRI session, the inflation level corresponding to the maximum tolerable volume (MTV) of a rectal balloon, was determined. The paradigm alternated three times a control period, where the balloon was completely deflated (l min), and a stimulus period (I min) where the balloon was inflated until 80% of the MTV. fMRI was performed at 1.5T (Philips all body, equipped with echoplanar imaging). Twenty five adjacent axial sections (4 mm thick), centred and parrallel to the bicommissural plane were measured 14 times during each period. An anatomic scan was performed at the end of the session to identify cerebral regions. Data were analyzed by SPM (statistical parametric mapping)software(4). Results: the group analysis (n=8) showed in IBS patients a) the absence of significant cerebral activation, b) significant desactivations (P
3161 A CONTROLLED INVESTIGATION OF THE EFFECTS OF A BRIEF RELAXATION REGIMEN ON THE SYMPTOMS OF IRRITABLE BOWEL SYNDROME. Miranda M. Baldwin, Val Heatley, St James's Univ Hosp, Leeds, United Kingdom. Despite many studies reporting significant links between stress and frequency of Irritable Bowel Syndrome (IBS) symptoms, currently there is little research on the use of relaxation as a realistic treatment method for IBS. This investigation concerned a controlled treatment trial of the effects of a brief, but specific relaxation regimen on Irritable Bowel Syndrome symptoms, all patients being seen within a gastroenterology out-patient clinic setting. A sample of 42 gastroenterology out-patients (74% female and mean duration of IBS symptoms was 62 months) were randomly allocated to either a study group of three half hour sessions of progressive and abdominal breathing relaxation, with education of stress and how it affects digestion, or to a control group of conventional treatment alone. Self report questionnaires incorporating the Manning criteria to measure IBS syptomology and HAD scales measuring mental well-being were used. The questionnaires were completed prior to treatment and three months post treatment. Patient compliance in the use of the relaxation techniques was encouraged via the use of a relaxation tape and recorded within a questionnaire, which revealed that patients were practising the abdominal breathing techniques regularly. The study group showed significantly greater (p value less than 0.0 I-using Mann-Whitney significance testing) improvement in overall Manning criteria symptoms following the relaxation treatment compared with the control group. Over 50% of the study group had shown improvemnts in IBS syptomology three months post treatment, particularly in pain and abdominal bloating, wheras the control group reported only a small improvement. Patients in both groups reported an increase in their HAD scores post treatment, with the study group reporting higher HAD scores than the control group but not significantly so. A high proportion of patients within the sample (66%) reported having had abdominal surgery, although it was not established whether this was prior or subsequent to IBS symptomology. This study reflects the ease of providing a reatively simple and non-invasive education based intervention that can be taught by any health professional in a gastroentology clinic setting. The findings suggest that the techniques used may provide an example of an cost-effective treatment for a chronically disabling condition, which clearly merits further larger scale investigation.
3163 LACTOBACILLUS PLANTARUM 299V IN THE IRRITABLE BOWEL SYNDROME: A RANDOMIZED, DOUBLE-BLIND, PLACEBO-CONTROLLED CROSSOVER STUDY. John K. DiBaise, John Lof, Karen Taylor, Eamonn M. Quigley, Univ of Nebraska Med Ctr, Omaha, NE. Recent evidence suggests that symptoms in the irritable bowel syndrome (IBS) may be related to either an exaggerated visceral sensitivity to normal luminal contents or to disturbances in the intraluminal milieu such as the intestinal flora, or to immune/inflammatory responses to intestinal infections. Lactobacillus plantarum 299V (LP299V), a probiotic agent with the ability to displace potentially pathogenic flora, has recently been shown, in a pilot study, to reduce the severity of abdominal pain in adults with IBS. Aim: To further characterize the effectiveness of LP299V in adults with IBS in a randomized, controlled setting. Methods: Subjects with at least a 6 month history of IBS, as defined by the Rome criteria, entered a 2 week run-in period on no IBS therapy. Randomly, they received either LP229V or placebo, in the form of juice, for 4 weeks followed by crossover to the alternate therapy for an additional 4 weeks. The two treatment periods were separated by a 2-week washout phase; and the study ended with a final 2 week washout phase off treatment. During the entire 14-week study period, IBS symptoms and bowel habits were recorded daily in a diary. Analysis of variance was used to determine whether there was a statistically significant treatment effect, order effect and interaction effect between treatment and order. Results: Twenty subjects completed the study (l7F; mean age 44.6). We found no significant differences between LP299V and placebo treatments or the order of treatments for the six outcome measurements: abdominal pain intensity, pain duration, bloating, number of bowel movements, stool characteristics or stool consistency. In addition, there was no significant carryover effect relating to the outcome measurements. Conclusion: In this small, randomized and controlled trial, we were unable to confirm reports of efficacy of LP299V in patients with IBS. Larger studies using alternative probiotics may be helpful to elucidate the role of probiotics in the management of IBS.
AGAA615
3160
3162
SULFASALAZINE IS A POTENT AND SPECIFIC INHIBITOR OF IKB-KINASES IKKA AND IKKB. Christoph Kurt Weber, Susanne Liptay, Thomas Wirth, Guido Adler, Roland Schmid.. Vniversit8t Ulm, Vim, Germany; Universitst, Vim, Germany; Univeritxt Vim, Ulm, Germany; Univ of Vim, Vim, Germany. Background & Aims: NF-KBlRel has been implicated in the pathogenesis of inflammatory bowel disease (IBD). Various drugs known to be effective for the treatment of IBD such as glucocorticoids and salicylates have been shown to interfere with NF-KBlRel signaling. The aim of this study was to define the molecular mechanism by which sulfasalazine inhibits NF-KB activation. Methods: The effects of sulfasalazine and its moieties on NF-KB signaling were evaluated using electromobility shift-, transfection- and immune complex kinase-assays. The direct effect of sulfasalazine on IKB kinase (IKK) activity was investigated using purified recombinant IKKaand IKKl3proteins. Results: NF-kBlRel activity induced by the overexpression of NF-KBinducing kinase (NIK), IKKa, IKKI3, or consitutively active IKKaand IKKl3mutants was inhibited in a dose dependent manner by sulfasalazine. Sulfasalazine inhibited TNFa, PMA and IL-I induced activation of endogenous IKKs in colonic epithelial and Jurkat T cells. Sulfasalazine did not affect other cell1ular kinases such as MAP-kinases JNK and p38. Sulfasalazine diminished the catalytic activity of purified IKKaand IKKl3in vitro. In contrast, the metabolic moieties of sulfasalazine, 5-aminosalicylic acid and sulfapyridine were not effective. The decrease in substrate phosphorylation by IKKaandl3is associated with a decrease of autophosphorylation of IKKs and can be antagonized by excess ATP. Conclusions: These data suggest that sulfasalazine is a competitive antagonist of ATP binding of IKKaand IKKI3. This suppression of NF-kB activation by inhibition of IKK activity may contribute to the well-known anti-inflammatory and immunosuppressive effects of sulfasalazine.
CENTRAL PROCESSING OF RECTAL PAIN IN IBS PATIENTS: AN FMRI STUDY. Bruno L. Bonaz, Emmanuel Papillon, Monica Baciu, Christophe Segebarth, Richard Bost, Jean-Francois Le Bas, Jacques Fournet, Dept of Gastroenterology, Grenoble, France; INSERM V438, Grenoble, France. Patients with irritable bowel syndrome (IBS) have a chronic visceral hyperalgesia as represented by hypersensitivity to balloon distension (I). Numerous arguments favour a role of the central nervous system in this hypersensitivity (2). Functional imaging techniques of the human brain, such as fMRI, present considerable interest, as they permit to depict local changes in brain activation during visceral sensitive or nociceptive stimulation. In healthy volunteers, using fMRI, we recently characterize cerebral loci activated by a rectal distension as for example the insular cortex, anterior cingulate cortex (ACC) and dorso-Iateral prefrontal cortex (DLPFC) (3). Aim: to characterize, using fMRI, cerebral loci activated by a rectal distension in IBS patients. Methods: 8 right-handed patients (7 women, I man, median: 42 years) were consecutively recruited. Prior to fMRI session, the inflation level corresponding to the maximum tolerable volume (MTV) of a rectal balloon, was determined. The paradigm alternated three times a control period, where the balloon was completely deflated (l min), and a stimulus period (I min) where the balloon was inflated until 80% of the MTV. fMRI was performed at 1.5T (Philips all body, equipped with echoplanar imaging). Twenty five adjacent axial sections (4 mm thick), centred and parrallel to the bicommissural plane were measured 14 times during each period. An anatomic scan was performed at the end of the session to identify cerebral regions. Data were analyzed by SPM (statistical parametric mapping)software(4). Results: the group analysis (n=8) showed in IBS patients a) the absence of significant cerebral activation, b) significant desactivations (P
3161 A CONTROLLED INVESTIGATION OF THE EFFECTS OF A BRIEF RELAXATION REGIMEN ON THE SYMPTOMS OF IRRITABLE BOWEL SYNDROME. Miranda M. Baldwin, Val Heatley, St James's Univ Hosp, Leeds, United Kingdom. Despite many studies reporting significant links between stress and frequency of Irritable Bowel Syndrome (IBS) symptoms, currently there is little research on the use of relaxation as a realistic treatment method for IBS. This investigation concerned a controlled treatment trial of the effects of a brief, but specific relaxation regimen on Irritable Bowel Syndrome symptoms, all patients being seen within a gastroenterology out-patient clinic setting. A sample of 42 gastroenterology out-patients (74% female and mean duration of IBS symptoms was 62 months) were randomly allocated to either a study group of three half hour sessions of progressive and abdominal breathing relaxation, with education of stress and how it affects digestion, or to a control group of conventional treatment alone. Self report questionnaires incorporating the Manning criteria to measure IBS syptomology and HAD scales measuring mental well-being were used. The questionnaires were completed prior to treatment and three months post treatment. Patient compliance in the use of the relaxation techniques was encouraged via the use of a relaxation tape and recorded within a questionnaire, which revealed that patients were practising the abdominal breathing techniques regularly. The study group showed significantly greater (p value less than 0.0 I-using Mann-Whitney significance testing) improvement in overall Manning criteria symptoms following the relaxation treatment compared with the control group. Over 50% of the study group had shown improvemnts in IBS syptomology three months post treatment, particularly in pain and abdominal bloating, wheras the control group reported only a small improvement. Patients in both groups reported an increase in their HAD scores post treatment, with the study group reporting higher HAD scores than the control group but not significantly so. A high proportion of patients within the sample (66%) reported having had abdominal surgery, although it was not established whether this was prior or subsequent to IBS symptomology. This study reflects the ease of providing a reatively simple and non-invasive education based intervention that can be taught by any health professional in a gastroentology clinic setting. The findings suggest that the techniques used may provide an example of an cost-effective treatment for a chronically disabling condition, which clearly merits further larger scale investigation.
3163 LACTOBACILLUS PLANTARUM 299V IN THE IRRITABLE BOWEL SYNDROME: A RANDOMIZED, DOUBLE-BLIND, PLACEBO-CONTROLLED CROSSOVER STUDY. John K. DiBaise, John Lof, Karen Taylor, Eamonn M. Quigley, Univ of Nebraska Med Ctr, Omaha, NE. Recent evidence suggests that symptoms in the irritable bowel syndrome (IBS) may be related to either an exaggerated visceral sensitivity to normal luminal contents or to disturbances in the intraluminal milieu such as the intestinal flora, or to immune/inflammatory responses to intestinal infections. Lactobacillus plantarum 299V (LP299V), a probiotic agent with the ability to displace potentially pathogenic flora, has recently been shown, in a pilot study, to reduce the severity of abdominal pain in adults with IBS. Aim: To further characterize the effectiveness of LP299V in adults with IBS in a randomized, controlled setting. Methods: Subjects with at least a 6 month history of IBS, as defined by the Rome criteria, entered a 2 week run-in period on no IBS therapy. Randomly, they received either LP229V or placebo, in the form of juice, for 4 weeks followed by crossover to the alternate therapy for an additional 4 weeks. The two treatment periods were separated by a 2-week washout phase; and the study ended with a final 2 week washout phase off treatment. During the entire 14-week study period, IBS symptoms and bowel habits were recorded daily in a diary. Analysis of variance was used to determine whether there was a statistically significant treatment effect, order effect and interaction effect between treatment and order. Results: Twenty subjects completed the study (l7F; mean age 44.6). We found no significant differences between LP299V and placebo treatments or the order of treatments for the six outcome measurements: abdominal pain intensity, pain duration, bloating, number of bowel movements, stool characteristics or stool consistency. In addition, there was no significant carryover effect relating to the outcome measurements. Conclusion: In this small, randomized and controlled trial, we were unable to confirm reports of efficacy of LP299V in patients with IBS. Larger studies using alternative probiotics may be helpful to elucidate the role of probiotics in the management of IBS.