COMPARATIVE STUDY OF THE MAIN FACTORS OF PATHOGENESIS OF INFLAMMATORY BOWEL DISEASE ON IN VITRO AND IN VIVO MODELS



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Abstract

BACKGROUND: Inflammatory bowel disease (IBD) is characterized by inflammation of the intestinal mucosa and increased intestinal permeability. In order to study the biological effects of drugs, it is important that experimental models adequately reproduce the main factors of pathogenesis of the disease.

AIM: To compare permeability of the intestinal epithelial barrier and the inflammation in models of IBD, based on lipopolysaccharide (LPS)-stimulated Caco-2 cells, and in mice with a knockout of the mucin 2 gene (Muc2-/-).

 MATERIALS AND METHODS: For in vitro IBD model, Caco-2 cells were exposed to LPS and assessed for transepithelial electrical resistance, monolayer permeability, expression of tight junction genes (ZO-1, Claudin-1) and pro-inflammatory cytokines (IL-8, TNF-α, IL-1β), and IL-8 secretion. Muc2-/- mice were used as an in vivo model of IBD. Intestinal permeability was determined by the concentration of fluorescein-isothiocyanate-dextran in the blood after its intragastric administration, a histological analysis of colon samples was performed, and the expression of the TNF-α, IL-1β, IL-10 and the content of IL-1β and IL-10 were assessed.

RESULTS: In vitro, 10 μg/ml LPS reduced transepithelial electrical resistance by 57%, increased the permeability of the cell monolayer by 38%, increased the expression of IL-8 and TNF-α by 2.8 and 2.3 times, decreased the expression of ZO-1 and Claudin-1 by 54 and 53%, increased the secretion of IL-8 by 27 times compared to the control. In vivo, intestinal permeability of Muc2-/- mice was 5.8 times higher, the expression of TNF-α, IL-1β was 9.9 and 6.8 times higher; IL-10 expression was 71% lower, intestinal levels of IL-1β and IL-10 were 94% and 44% lower compared to healthy mice.

 CONCLUSIONS: In vitro and in vivo models of IBD are characterized by similar dynamics in intestinal permeability and inflammatory response. These models adequately reproduce the pathogenesis of IBD and complement each other.

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About the authors

Tatiana S. Sall

Institute of Experimental Medicine

Author for correspondence.
Email: miss_taty@mail.ru
ORCID iD: 0000-0002-5890-5641
SPIN-code: 4172-6277
Scopus Author ID: 57194050387

Research Associate, Department of Biochemistry

Russian Federation, 12 Academician Pavlov St., Saint Petersburg, 197022, Russia

Ekaterina A. Litvinova

Novosibirsk State Technical University

Email: dimkit@mail.ru
ORCID iD: 0000-0001-6398-7154
SPIN-code: 2995-8611
Scopus Author ID: 7005626124

Cand. Sci. (Biol.), Research Associate

Russian Federation, 20 K. Marx Ave., Novosibirsk, 630073, Russia

Elena L. Arzhanova

Novosibirsk State University

Email: e.arzhanova@g.nsu.ru
ORCID iD: 0009-0006-1066-1867
Scopus Author ID: 58569546100

student

Russian Federation, 1 Pirogova str., Novosibirsk, 630090, Russia

Tatiana A. Kashina

Peter the Great St. Petersburg Polytechnic University

Email: tat.kashina@list.ru
ORCID iD: 0000-0002-7314-8298
SPIN-code: 4713-4128

Student

Russian Federation, 29 B Polytechnicheskaya, Saint Petersburg, 195251, Russia

Irina V. Voronkina

Institute of Experimental Medicine

Email: voronirina@list.ru
ORCID iD: 0000-0003-0078-4442
SPIN-code: 2336-4158
Scopus Author ID: 6602535489

Cand. Sci. (Biol.), Senior Research Associate, Department of Biochemistry

Russian Federation, 12 Academician Pavlov St., Saint Petersburg, 197022, Russia

Olga V. Kirik

Institute of Experimental Medicine

Email: olga_kirik@mail.ru
ORCID iD: 0000-0001-6113-3948
SPIN-code: 5725-8742
Scopus Author ID: 27171304100

Cand. Sci. (Biol.), Senior Research Associate, Department of General and Private Morphology

Russian Federation, 12 Academician Pavlov St., Saint Petersburg, 197022, Russia

Stanislav I. Sitkin

Almazov National Medical Research Centre;
North-Western State Medical University Named after I.I. Mechnikov

Email: sitkins@yandex.ru
ORCID iD: 0000-0003-0331-0963
SPIN-code: 3961-8815
Scopus Author ID: 6603071466

MD, Cand. Sci. (Med.), Head of the research group of epigenetics and metagenomics in perinatology and pediatrics; Associate Professor of the Department of Propaedeutics of Internal Diseases, Gastroenterology and Dietetics 

Russian Federation, 2 Akkuratova street, Saint Petersburg, 197341, Russia; 47 Piskarevskij prospect, Saint Petersburg, 195067, Russia

Timur Ya. Vakhitov

Institute of Experimental Medicine

Email: tim-vakhitov@yandex.ru
ORCID iD: 0000-0001-8221-6910
SPIN-code: 7298-2571
Scopus Author ID: 55406626900

Dr. Sci. (Biol.), Chief Researcher of Non-Infectious Disease Metabolomics Group

Russian Federation, 12 Academician Pavlov St., Saint Petersburg, 197022, Russia

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