vavilovВавиловский журнал генетики и селекцииVavilov Journal of Genetics and Breeding2500-3259Institute of Cytology and Genetics of Siberian Branch of the RAS10.18699/VJ15.066vavilov-441Research ArticleМОДЕЛИРОВАНИЕ ПАТОЛОГИЙMODELING OF DISORDERS AND EXPERIMENTAL TREATMENTВклад кишечного муцина-2 в эффективность антибактериальной терапии Helicobacter spp. у лабораторных мышейRole of intestinal mucin-2 in the effectiveness of the treatment of Helicobacter spp. infection in laboratory miceЛитвиноваЕ. А.LitvinovaE. A.litvinova@bionet.nsc.ruБеляевМ. Д.BelyaevM. D.ПрохорчукА. В.ProkhortchoukA. V.КоростинаВ. С.KorostinaV. S.ПрохорчукЕ. Б.ProkhortchoukE. B.КожевниковаЕ. Н.KozhevnikovaE. N.kozhevnikova@bionet.nsc.ruФедеральное государственное бюджетное научное учреждение «Федеральный исследовательский центр Институт цитологии и генетики Сибирского отделения Российской академии наук», Новосибирск, РоссияРоссияInstitute of Cytology and Genetics SB RAS, Novosibirsk, RussiaRussian FederationИнститут биоинженерии, Федеральный исследовательский центр «Фундаментальные основы биотехнологии» Российской академии наук, Москва, РоссияРоссияInstitute of Bioengineering, Research Center of Biotechnology of the Russian Academy of Sciences, Moscow, RussiaRussian Federation201501122015194494498Copyright © Литвинова Е.А., Беляев М.Д., Прохорчук А.В., Коростина В.С., Прохорчук Е.Б., Кожевникова Е.Н., 20152015Литвинова Е.А., Беляев М.Д., Прохорчук А.В., Коростина В.С., Прохорчук Е.Б., Кожевникова Е.Н.Litvinova E.A., Belyaev M.D., Prokhortchouk A.V., Korostina V.S., Prokhortchouk E.B., Kozhevnikova E.N.This work is licensed under a Creative Commons Attribution 4.0 License.https://vavilov.elpub.ru/jour/article/view/441

Нарушение синтеза основного протеогликана кишечника (муцин-2) у людей характерно как при развитии неспецифического язвенного колита, так и при болезни Крона. Такие морфологические изменения кишечного слизистого слоя могут повлиять на разнообразие кишечной микрофлоры. Использование антибиотиков для лечения бактериальных инфекций у людей и животных с нарушением синтеза муцина-2 может быть малоэффективным, а иногда и опасным из-за развития сепсиса или хронического воспаления. В работе на мышах с генетически детерминированным дефицитом муцина-2 (линия Muc2–/–) исследовали как применение трех антибиотиков (кларитромицин, амоксицилин и метронидазол) влияет на элиминацию патогенной инфекции у мышей. Для оценки эффективности антибиотиков измеряли количество патогенных (Helicobacter spp.) и симбиотических (E. coli) бактерий в кишечнике животных. Негативное влияние антибактериальной терапии на организм хозяина оценивали по выживаемости животных. Три антибиотика не способствовали устранению Helicobacter spp. у мышей с дефицитом муцина-2. Наряду с неэффективным лечением была отмечена гибель у 60 % животных данной линии. Мыши с нормальной функцией синтеза муцина-2 (C57BL/6J) имели 100 %-е выживание. Животные с нормальной функцией синтеза основного кишечного протеогликана не демонстрировали снижения массы тела. Helicobacter spp. был полностью устранен у этих животных. Таким образом, лечение инфекции Helicobacter spp. антибиотиками у животных с нарушением синтеза муцина-2 не только малоэффективно, но и вызывает гибель животного. Высокая восприимчивость к токсическому действию антибиотиков дает основание для использования мышей с дефицитом муцина-2 в качестве тест-объекта для оценки фармакологической безопасности новых средств антибактериальной терапии.

Abnormal synthesis of the main intestinal proteo­glycan mucin-2 is typical of ulcerative colitis and Crohn’s disease in humans. Those morphological changes of the mucus layer affect the diversity of the intestinal microflora. Antibiotics may be ineffective or even dangerous to humans or animals deficient for mucin-2 because of the risk of sepsis and chronic inflammation. In this study, we investigated the potential of antibiotics (clarithromycin, amoxicillin, and metronidazole) in elimination of patho­genic infection from Muc2 knockout mice (Muc2–/–). We assayed the population sizes of pathogens (Heli­co­bacter spp.) and symbiotic (E. coli) bacteria in the intestines of animals as a criterion of antibiotic efficacy. The damaging effect of antibacterial treatment on the host body was estimated from their survival rate. Three antibiotics were ineffective in the elimination of Helicobacter spp. from mucin-2-deficient mice. Moreover, the mortality of Muc2 knockout mice during the antibacterial treatment was 60 %. The survival of wild-type mice (C57BL/6J) during the treatment was 100 %. The weight of wild-type mice showed no decrease during the treatment. The Helico­bacter spp. pathogen was fully eradicated from wild-type mice. Thus, therapy of Helicobacter spp. infection in mucin-2 deficient animals is not only poorly efficient but even deadly. The high susceptibility to antibiotics allows Muc2 knockout mice to be used as a test model to evaluate the pharmacological safety of new antibiotics.

мышикишечное воспалениемуцин-2антибактериальная терапияmiceintestinal inflammationmucin-2antibacterial treatmentРФФИReferencesAyres J.S., Trinidad N.J., Vance R.E. Lethal inflammasome activation by a multidrug-resistant pathobiont upon antibiotic disruption of the microbiota. Nat. Med. 2012;18(5):799-806. DOI: 10.1038/nm.2729Ayres J.S., Trinidad N.J., Vance R.E. Lethal inflammasome activation by a multidrug-resistant pathobiont upon antibiotic disruption of the microbiota. Nat. Med. 2012;18(5):799-806. DOI: 10.1038/nm.2729Bergstrom K.S.B., Kissoon-Singh V., Gibson D.L., Ma C., Montero M., Sham H.P., Vallance B.A. Muc2 protects against lethal infectious colitis by disassociating pathogenic and commensal bacteria from the colonic mucosa. PLoS Pathog. 2010;6(5):e1000902. DOI: 10.1371/journal.ppat.1000902Bergstrom K.S.B., Kissoon-Singh V., Gibson D.L., Ma C., Montero M., Sham H.P., Vallance B.A. Muc2 protects against lethal infectious colitis by disassociating pathogenic and commensal bacteria from the colonic mucosa. PLoS Pathog. 2010;6(5):e1000902. DOI: 10.1371/journal.ppat.1000902Bulois P., Desreumaux P., Neut C., Da A., Cortot A., Colombel J.F. Infectious agents and Crohn’s disease. Clin. Microbiol. Infect. 1999;5(10):601-604. DOI: 10.1111/j.1469-0691.1999.tb00415.xBulois P., Desreumaux P., Neut C., Da A., Cortot A., Colombel J.F. Infectious agents and Crohn’s disease. Clin. Microbiol. Infect. 1999;5(10):601-604. DOI: 10.1111/j.1469-0691.1999.tb00415.xChin E.Y., Dangler C.A., Fox J.G., Schauer D.B. Helicobacter hepaticus infection triggers inflammatory bowel disease in T cell receptor alphabeta mutant mice. Comparative Med. 2000;50(6):586-594.Chin E.Y., Dangler C.A., Fox J.G., Schauer D.B. Helicobacter hepaticus infection triggers inflammatory bowel disease in T cell receptor alphabeta mutant mice. Comparative Med. 2000;50(6):586-594.Ghai O.P., Menon P.S.N., Bhau M.K. Pathogenesis of diarrhea due to Escherichia coli. Indian J. Pediatr. 1980;47(4):311-316. DOI: 10.1007/BF02831325Ghai O.P., Menon P.S.N., Bhau M.K. Pathogenesis of diarrhea due to Escherichia coli. Indian J. Pediatr. 1980;47(4):311-316. DOI: 10.1007/BF02831325Hansson G.C., Johansson M.E.V. The inner of the two Muc2 mucin-dependent mucus layers in colon is devoid of bacteria. Gut. Microbes. 2010;1(1):51-54. DOI: 10.4161/gmic.1.1.10470Hansson G.C., Johansson M.E.V. The inner of the two Muc2 mucin-dependent mucus layers in colon is devoid of bacteria. Gut. Microbes. 2010;1(1):51-54. DOI: 10.4161/gmic.1.1.10470Kamada N., Chen G.Y., Inohara N., Núñez G. Control of pathogens and pathobionts by the gut microbiota. Nat. Immunol. 2013;14(7):685690. DOI: 10.1038/ni.2608Kamada N., Chen G.Y., Inohara N., Núñez G. Control of pathogens and pathobionts by the gut microbiota. Nat. Immunol. 2013;14(7):685-690. DOI: 10.1038/ni.2608Kerton A., Warden P. Review of successful treatment for Helicobacter species in laboratory mice. Lab. Anim. 2006;40(2):115-122. DOI: 10.1258/002367706776319033Kerton A., Warden P. Review of successful treatment for Helicobacter species in laboratory mice. Lab. Anim. 2006;40(2):115-122. DOI: 10.1258/002367706776319033Martens E.C., Koropatkin N.M., Smith T.J., Gordon J.I. Complex glycan catabolism by the human gut microbiota: The bacteroidetes sus-like paradigm. J. Biol. Chem. 2009;284(37):24673-24677. DOI: 10.1074/jbc.R109.022848Martens E.C., Koropatkin N.M., Smith T.J., Gordon J.I. Complex glycan catabolism by the human gut microbiota: The bacteroidetes sus-like paradigm. J. Biol. Chem. 2009;284(37):24673-24677. DOI: 10.1074/jbc.R109.022848Martino-Cardona M. del C., Beck S.E., Brayton C., Watson J. Eradication of Helicobacter spp. by using medicated diet in mice deficient in functional natural killer cells and complement factor D. J. Am. Assoc. Lab. Anim. Sci. 2010;49(3):294-299.Martino-Cardona M. del C., Beck S.E., Brayton C., Watson J. Eradication of Helicobacter spp. by using medicated diet in mice deficient in functional natural killer cells and complement factor D. J. Am. Assoc. Lab. Anim. Sci. 2010;49(3):294-299.Monceaux C.P., Testerman T.L., Boktor M., Jordan P., Adegboyega P., Mcgee D.J., Alexander J.S. Helicobacter infection decreases basal colon inflammation, but increases disease activity in experimental IBD. Open J. Gastroenterol. 2013;3:177-189. DOI: 10.4236/ojgas.2013.33029Monceaux C.P., Testerman T.L., Boktor M., Jordan P., Adegboyega P., Mcgee D.J., Alexander J.S. Helicobacter infection decreases basal colon inflammation, but increases disease activity in experimental IBD. Open J. Gastroenterol. 2013;3:177-189. DOI: 10.4236/ojgas.2013.33029Morgan X.C., Tickle T.L., Sokol H., GeversD., Devaney K.L., Ward D.V., Huttenhower C. Dysfunction of the intestinal microbiome in inflammatory bowel disease and treatment. Genome Biol. 2012;13(9):R79. DOI: 10.1186/gb-2012-13-9-r79Morgan X.C., Tickle T.L., Sokol H., Gevers D., Devaney K.L., Ward D.V., Huttenhower C. Dysfunction of the intestinal microbiome in inflammatory bowel disease and treatment. Genome Biol. 2012;13(9):R79. DOI: 10.1186/gb-2012-13-9-r79Nicklas W., Baneux P., Boot R., Decelle T., Deeny A.A., Fumanelli M., Illgen-Wilcke B. Recommendations for the health monitoring of rodent and rabbit colonies in breeding and experimental units. Lab. Anim. 2002;36:20-42.Nicklas W., Baneux P., Boot R., Decelle T., Deeny A.A., Fumanelli M., Illgen-Wilcke B. Recommendations for the health monitoring of rodent and rabbit colonies in breeding and experimental units. Lab. Anim. 2002;36:20-42.Nicklas W., Deeny A., DiercksP., Gobbi A., Illgen-WilckeB., SeidelinM. FELASA guidelines for the accreditation of health monitoring programs and testing laboratories involved in health monitoring. Lab. Anim. 2010;39(2):43-48. DOI: 10.1258/la.2009.009086Nicklas W., Deeny A., Diercks P., Gobbi A., Illgen-Wilcke B., Seidelin M. FELASA guidelines for the accreditation of health monitoring programs and testing laboratories involved in health monitoring. Lab. Anim. 2010;39(2):43-48. DOI: 10.1258/la.2009.009086O’Connor M. Ulcerative Colitis – Treatments, Special Populations and the Future. Publ.: InTech, 2011. DOI: 10.5772/2369O’Connor M. Ulcerative Colitis – Treatments, Special Populations and the Future. Publ.: InTech, 2011. DOI: 10.5772/2369Pilarczyk-Zurek M., Chmielarczyk A., Gosiewski T., Tomusiak A., Adamski P., Zwolinska-Wcislo M., Strus M. Possible role of Escherichia coli in propagation and perpetuation of chronic inflammation in ulcerative colitis. BMC Gastroenterol. 2013;13(1):61. DOI: 10.1186/1471-230X-13-61Pilarczyk-Zurek M., Chmielarczyk A., Gosiewski T., Tomusiak A., Adamski P., Zwolinska-Wcislo M., Strus M. Possible role of Escherichia coli in propagation and perpetuation of chronic inflammation in ulcerative colitis. BMC Gastroenterol. 2013;13(1):61. DOI: 10.1186/1471-230X-13-61Rakoff-Nahoum S., Medzhitov R. Regulation of spontaneous intestinal tumorigenesis through the adaptor protein MyD88. Science. 2007;317(5834):124-127. DOI: 10.1126/science.1140488Rakoff-Nahoum S., Medzhitov R. Regulation of spontaneous intestinal tumorigenesis through the adaptor protein MyD88. Science. 2007;317(5834):124-127. DOI: 10.1126/science.1140488Sharp J.M., Vanderford D.A., Chichlowski M., Myles M.H., Hale L.P. Helicobacter infection decreases reproductive performance of IL10deficient mice. Comparative Med. 2008;58(5):447-453.Sharp J.M., Vanderford D.A., Chichlowski M., Myles M.H., Hale L.P. Helicobacter infection decreases reproductive performance of IL10-deficient mice. Comparative Med. 2008;58(5):447-453.Shomer N.H., Dangler C.A., Marini R.P., Fox J.G. Helicobacter bilis/ Helicobacter rodentium co-infection associated with diarrhea in a colony of SCID mice. Lab. Anim. Sci. 1998;48(5):455–459.Shomer N.H., Dangler C.A., Marini R.P., Fox J.G. Helicobacter bilis/ Helicobacter rodentium co-infection associated with diarrhea in a colony of SCID mice. Lab. Anim. Sci. 1998;48(5):455–459.Ubeda C., Taur Y., Jenq R.R., Equinda M.J., Son T., Samstein M., Pamer E.G. Vancomycin-resistant Enterococcus domination of intestinal microbiota is enabled by antibiotic treatment in mice and precedes bloodstream invasion in humans. J. Clin. Invest. 2010;120(12):43324341. DOI: 10.1172/JCI43918Ubeda C., Taur Y., Jenq R.R., Equinda M.J., Son T., Samstein M., Pamer E.G. Vancomycin-resistant Enterococcus domination of intestinal microbiota is enabled by antibiotic treatment in mice and precedes bloodstream invasion in humans. J. Clin. Invest. 2010;120(12):4332-4341. DOI: 10.1172/JCI43918Van der Sluis M., de Koning B.A.E., de Bruijn A.C.J.M., Velcich A., Meijerink J.P.P., van Goudoever J.B., Einerhand A.W.C. Muc2-deficient mice spontaneously develop colitis, indicating that MUC2 is critical for colonic protection. Gastroenterology. 2006;131(1):117129. DOI: 10.1053/j.gastro.2006.04.020Van der Sluis M., de Koning B.A.E., de Bruijn A.C.J.M., Velcich A., Meijerink J.P.P., van Goudoever J.B., Einerhand A.W.C. Muc2-deficient mice spontaneously develop colitis, indicating that MUC2 is critical for colonic protection. Gastroenterology. 2006;131(1):117-129. DOI: 10.1053/j.gastro.2006.04.020Wlodarska M., WillingB., Keeney K.M., Menendez A., Bergstrom K.S., Gill N., Finlay B.B. Antibiotic treatment alters the colonic mucus layer and predisposes the host to exacerbated Citrobacter rodentium-induced colitis. Infect. Immun. 2011;79(4):1536-1545. DOI: 10.1128/IAI.01104-10Wlodarska M., Willing B., Keeney K.M., Menendez A., Bergstrom K.S., Gill N., Finlay B.B. Antibiotic treatment alters the colonic mucus layer and predisposes the host to exacerbated Citrobacter rodentium-induced colitis. Infect. Immun. 2011;79(4):1536-1545. DOI: 10.1128/IAI.01104-10

The authors declare that there are no conflicts of interest present.