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  • Inflammatory Disorders

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    Inflammatory Disorders

    Загружено:

    api-3714923

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    Attribution Non-Commercial (BY-NC)
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    Notion Rete. V7, No. pp 46D-473, 1957 ‘Copygh © 199 Skee Scere Ina Prot be USA. al gh eer consist si0.00% 00 PI $0271-5317(97)00007.9 GLUTATHIONE LEVELS IN CHRONIC INFLAMMATORY DISORDERS (OF THE HUMAN COLON Eduardo A. Ruan, M.D,,Simbasiva Rao, M.D., J. Steven Burdick, MD., Steven J. Suyker, MD., Gordon L. Telford, M.D, Mary F. Ouersoa, MD. Emmanuel C. Opara, PAD., and Timotiy R. Koch, MD* Departments of Medicine and Surgery, Zablocki VA Medical Center, Medical College of Wisconsin, Milwaukes, Wisconsin; Departments of Surgery and Pathology, Northwestern University Medical School, Chicago, Ilinois; and Duke University Medical Center, Durham, North Carolina ‘ABSTRACT Giutathone depletion has beon described in tissues obtained from several ehronie diseases. Tereased fre radical production by inflammatory cells occurs in inflammatory bowel disease. ‘We hypothesized that this could induce depletion of gut antioxidants. In this study, we examined the potential relationship between chronic inflammation and colonic glutathione levels. Using a validated assay, glutathione levels were determined inthe mucosal-submucosal layer and the muscularis extema layer in surgical colonie specimens from 26 patients with ulcerative colitis, 14 patients with Crohn's clits, and 10 patients who underwent partial colozomy fornoa-obstructve ncplasia, Inflammation was graded histologically. Glutathione levels were decreaod inthe muscularis extera and inthe mucosal-submucosal layers from both loerative colitis and Cros colitis (both p<0.05) There were parallel declines of glutathione levels in the muscularis extema layer compared to the mucosal-submucosal layer from individual eolonie specimens. In ulcerative eois, glutathione levels were reduced in histologealy setive disease compared to inactive diseae (in the mucosal-submucosal layers: ‘MeantSEM were 214468 nmol/g wet tissue and 808250, respectively; in the muscularis, ‘externa layers: 333497 and 8905340; both p<0.05). In Cru ols, Uhre were no significant differences between histologically active and inactive disease (inthe mucosal-submucosal layers: 114853 and 4614206; in the muscularis extema layers: 10559 and 5532211; both p>0.05). This study provides evidence that chronic inflammatory disorders ofthe colon are ‘associated with gllathione depletion. In lesrative colt, thre was a relationship betwesa the severity of inflammation and glutathione depletion. By contrast, this relationship was not significant in Croba's colts. The results suggest that there could be a primary defect in _slutathione production in Crohn's clits, or a difference in the relative levels of fee radical ‘production by inflammatory ells present in these two disorders of colonic inflammation. api 81 Bar Seo, Key Words: Glutathione, Colon, Crofn's disease, Ulcerative colitis, Antioxidant, Inflammatory bowel disease. ‘commending Aur Di Tenth R. Kosh Chie Seton of Castorlerclgy, Robt ©. Byrd Heath Ses (Centr of West Vig Unveriy, P0, Box 9161, Mogutown, Wes Virgina 26506-9161, 466 E.A RUAN etal INTRODUCTION GGitathione i a teipeptide sil present in most mammalian ells Ithas bea reported that tissue levels. of this non-enzymatic antioxidant are depleted in several ehronie disorders including iiopathic pulmonary fibrosis, human immunodetciency virw-tlated disease, and respiratory disuess syndrome (I). Wentfiction of acquired glutathione deficiency states could be helpful in predicting the poteatial benefit of glutathione supplementation in human diseases, Fre radial production has neg sa common pathway of tissue injury in a arey of diseass. There is evidence supporting ineeased fv eicalprodution by inlaratory cell in he intial mucosa Com patcnts with conic iflaralory bowel disease (2). Unfortunatly, ithas ben dficl to aseiminate bsween a tialgi ole for fee rails as opposed to heir presence a a elletion of jury caused by oer potential agents (6. ‘Mammalian tissues contain a munber of diferent fee radical seavenging systems including enzymes (i, catalase), eopper/Zine (CuZA) containing proteins (ie, superoxide dismutase) and tissue micronutrients, such as ascorbate and glutathione, Ther is 8 paucy of information available shout the status ofthese defenses in colon obtained from chronic inflammatory conditions (7-9). Decreased glutathione levels have been reported in the terminal ileal mucosa from patists with active Crohn's disease compared to areas withowt setive inflammation and compared to contols without inflammatory bowel disease. This suggests increased ‘consumption ofthis antioxidant bythe inflammtory process (8). thos been proposed that the therapeutic effet of -aminosaliylic acid in the inflammatory bowel diseases is related to its free radical scavenging action (J0), There has been interest in developing new pharmacological approaches forthe conizol of tissue damage induced by fre radial production in the eronic {inflammatory bowel diseases. Uconlied trials of superoxide dismutase have shown some promise (1) lever, his enzyme requires a special dlvery system tat pemit i to reach its therapeutic target before being destroyed. Since glutathione, given orally, is absorbed intact by the intestine (12), i constitutes an ataetive possiblity for Unerapeuti trials i inflammatory bowel disease, Based on these previous sues, we ypothesiad that inereased fre radical production by inflammatory cellin the inflammatory bowel diseases could induce colonic depletion ofthe antioxidant, glutathione. In this study, we measured sue glutathione levels in sugical specimens of elon fom patients with ulcerative colitis and Croka's colitis, and from patients with non-obstvetng colonic neoplasia as controls. The presence of and severity of inflammation was determined by histopathology in order to examine the potential relationship between inflammation and colonic glutathione levels METHODS Tisue Specimens, Ths say was agroved by he Human Research Review Corus ofthe Medel Coleg of Wisconsin, Colonic specimens were clined at surgry from pants with Cros cols (n= 1), {erative cols (n= 26a in pans vir or ruc clone neoplasia who seved a eontcs (n= 10). ‘Among controls, pcinens Of gostynomal exons sss were otsned fom he animes bord feast 3 cm distance rom the tumor margin nd fom the taped ends, Coli specimens were extracted ar ‘eid as posible by clanging te ood ply oe colon before removal om he abdominal cavity in cer to minimize ichenia, All specimens were quek-Gozea and stored at-76"C. The mucosa-submicas and muscularis externa laycs wee separated pit o assy by microdissection in ice-cold 09% NACL GLUTATHIONE LEVELS IN HUMAN COLON CELLS. 485 Glutathione Determination, Extracts of each layer were propared by homogenizing tissue in 2 suspension 431% slfosalclic acd containing 0.25% Disodiur-EDTA, and then centrifuging at 8,000 g foe 30 minutes a 4° C, Determination of glutathione levels was performed as previously deseribed (13) using the euler spetrophotomctic assay as modified by Lang (/¢). leis known that issue protein levels ae higher in the gut wall in Crohn's disease (/5). In prior studies of lyophilized colonic tissue, we have shown that normal and abnormal (ulcerative colts and Croti's disease) colonic specimens did not differ in water content (14), Consequently, glutathione results in this present study were expressed as nmoles/g wet tissue. Histology. Transmsal soctions ofthe specimens were stained with the hematoxylin and eosin metho, ‘nd then examined by apatbologist blinded). On histologic examination, inflammation ia tissues fom patients ‘ith Cob’ disease was classed as inactive" and "active and in tissues from paticns with ulcerative colitis a8 “inactive”, “mild or moderately active" of "severely active", using standardized eriteria (17), Staistcal Analysis, Quantitative variables were expressed as Meant SEM, Differences in glutathione levels between the groups were studied using non-parametric analysis (Kruskal-Wallis one-way ANOVA on ‘anks), and all pairwise multiple comparisons were then earied out using Dunn's method. To examine the potential lationship between glusthione levels inthe to colene layer, linear corelation analysis was utilized, ‘Thereationships betwon tissu lution lovels and patents age or duration of disease were ested by Pearson coreation, Ap value

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