Name: investigating intestinal inflammation in dss-induced model of ibd
Uploaded: 2012-02-01T13:00:00
Description: Watch this Scientific Journal Video about Investigating Intestinal Inflammation in DSS-induced Model of IBD at JoVE.com

Работа выполнена при поддержке грантов от канадского Института Здоровья Исследований (CIHR) и Крона и колит Фонд Канады (CCFC).

1. Мышиной модели DSS-индуцированного острого колита <ol><li> Добавить декстран сульфат натрия (DSS) для автоклавного питьевой водой желаемой конечной концентрации (1-5%) (вес / объем) (т. е. Для того, чтобы 5% DSS решение, добавьте 25 мг DSS порошка на 500 мл воды из автоклавного) . Маточный раствор можно ?...

<ol>
	<li>Sands, B. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=From+symptom+to+diagnosis:+clinical+distinctions+among+various+forms+of+intestinal+inflammation.">From symptom to diagnosis: clinical distinctions among various forms of intestinal inflammation.</a> Gastroenterology. 126, 1518-1532 (2004).</li><li>Danese, S., Fiocchi, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Etiopathogenesis+of+inflammatory+bowel+diseases.">Etiopathogenesis of inflammatory bowel diseases.</a> World J. Gastroenterol. 12, 4807-4812 (2006).</li><li>Wirtz, S., Neufert, C., Weigmann, B., Neurath, M. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemically+induced+mouse+models+of+intestinal+inflammation.">Chemically induced mouse models of intestinal inflammation.</a> Nat. Protoc. 2, 541-546 (2007).</li><li>Axelsson, L. G., Landstrom, E., Goldschmidt, T. J., Gronberg, A., Bylund-Fellenius, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dextran+sulfate+sodium+(DSS)+induced+experimental+colitis+in+immunodeficient+mice:+effects+in+CD4(+)-cell+depleted,+athymic+and+NK-cell+depleted+SCID+mice.">Dextran sulfate sodium (DSS) induced experimental colitis in immunodeficient mice: effects in CD4(+)-cell depleted, athymic and NK-cell depleted SCID mice.</a> Inflamm. Res. 45, 181-191 (1996).</li><li>Egger, B., Bajaj-Elliott, M., MacDonald, T. T., Inglin, R., Eysselein, V. E., Büchler, M. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterisation+of+acute+murine+dextran+sodium+sulphate+colitis:+cytokine+profile+and+dose+dependency.">Characterisation of acute murine dextran sodium sulphate colitis: cytokine profile and dose dependency.</a> Digestion. 62, 240-248 (2000).</li><li>Krawisz, J. E., Sharon, P., Stenson, W. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+assay+for+acute+intestinal+inflammation+based+on+myeloperoxidase+activity.+Assessment+of+inflammation+in+rat+and+hamster+models.">Quantitative assay for acute intestinal inflammation based on myeloperoxidase activity. Assessment of inflammation in rat and hamster models.</a> Gastroenterology. 87, 1344-1350 (1984).</li><li>Cooper, H. S., Murthy, S. N., Shah, R. S., Sedergran, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinicopathologic+study+of+dextran+sulfate+sodium+experimental+murine+colitis.">Clinicopathologic study of dextran sulfate sodium experimental murine colitis.</a> Lab. Invest. 69, 238-249 (1993).</li><li>Smith, J. W., Castro, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Relation+of+peroxidase+activity+in+gut+mucosa+to+inflammation.">Relation of peroxidase activity in gut mucosa to inflammation.</a> Am. J. Physiol. 234, R72-R79 (1978).</li><li>Ghia, J. E., Blennerhassett, P., Kumar-Ondiveeran, H., Verdu, E. F., Collins, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+vagus+nerve:+a+tonic+inhibitory+influence+associated+with+inflammatory+bowel+disease+in+a+murine+model.">The vagus nerve: a tonic inhibitory influence associated with inflammatory bowel disease in a murine model.</a> Gastroenterology. 131, 1122-1130 (2006).</li><li>Okayasu, I., Hatakeyama, S., Yamada, M., Ohkusa, T., Inagaki, Y., Nakaya, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+method+in+the+induction+of+reliable+experimental+acute+and+chronic+ulcerative+colitis+in+mice.">A novel method in the induction of reliable experimental acute and chronic ulcerative colitis in mice.</a> Gastroenterology. 98, 694-702 (1990).</li><li>Solomon, L., Mansor, S., Mallon, P., Donnelly, E., Hoper, M., Loughrey, M., Kirk, S., Gardiner, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+dextran+sulphate+sodium+(DSS)+model+of+colitis:+an+overview.">The dextran sulphate sodium (DSS) model of colitis: an overview.</a> Comparative Clinical Pathology. 19, 235-239 (2010).</li><li>Mähler, M., Bristol, I. J., Leiter, E. H., Workman, A. E., Birkenmeier, E. H., Elson, C. O., Sundberg, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+susceptibility+of+inbred+mouse+strains+to+dextran+sulfate+sodium-induced+colitis.">Differential susceptibility of inbred mouse strains to dextran sulfate sodium-induced colitis.</a> Am. J. Physiol. 274, G544-G551 (1998).</li><li>Hans, W., Scholmerich, J., Gross, V., Falk, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+the+resident+intestinal+flora+in+acute+and+chronic+dextran+sulfate+sodium-induced+colitis+in+mice.">The role of the resident intestinal flora in acute and chronic dextran sulfate sodium-induced colitis in mice.</a> Eur. J. Gastroenterol. Hepatol. 12, 267-273 (2000).</li><li>Rath, H. C., Schultz, M., Freitag, R., Dieleman, L. A., Li, F., Linde, H., Schölmerich, J., Sartor, R. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Different+subsets+of+enteric+bacteria+induce+and+perpetuate+experimental+colitis+in+rats+and+mice.">Different subsets of enteric bacteria induce and perpetuate experimental colitis in rats and mice.</a> Infect. Immun. 69, 2277-2285 (2001).</li><li>Ghia, J. E., Li, N., Wang, H., Collins, M., Deng, Y., El-Sharkawy, R. T., Côté, F., Mallet, J., Khan, W. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Serotonin+has+a+key+role+in+pathogenesis+of+experimental+colitis.">Serotonin has a key role in pathogenesis of experimental colitis.</a> Gastroenterology. 137, 1649-1660 (2009).</li><li>Elson, C. O., Beagley, K. W., Sharmanov, A. T., Fujihashi, K., Kiyono, H., Tennyson, G. S., Cong, Y., Black, C. A., Ridwan, B. W., McGhee, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hapten-induced+model+of+murine+inflammatory+bowel+disease:+mucosa+immune+responses+and+protection+by+tolerance.">Hapten-induced model of murine inflammatory bowel disease: mucosa immune responses and protection by tolerance.</a> J. Immunol. 157, 2174-2185 (1996).</li><li>Dieleman, L. A., Ridwan, B. U., Tennyson, G. S., Beagley, K. W., Bucy, R. P., Elson, C. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dextran+sulfate+sodium-induced+colitis+occurs+in+severe+combined+immunodeficient+mice.">Dextran sulfate sodium-induced colitis occurs in severe combined immunodeficient mice.</a> Gastroenterology. 107, 1643-1652 (1994).</li></ol>

DSS колит широко используются химически индуцированных модель кишечного воспаления. В этой модели мышей приведены питьевой воды дополнена DSS, который, как полагают, быть токсичными для клеток эпителия кишечника и нарушают целостность слизистой оболочки барьер. 10 Администрации DSS ...

<table border="1"><tbody><tr><td> Название реагента / оборудование: </td><td> Компания </td><td> Номер по каталогу </td><td> Комментарии </td></tr><tr><td> Эппендорф Безопасный-Lock Микроцентрифуга Тюбе (2 мл) </td><td> Эппендорф </td><td> 0030 120.094 </td><td></td></tr><tr><td> Biotek EL808 Поглощение ридер </td><td> Биотек </td><td> EL808 </td><td></td></tr><tr><td> Декстран сульфата натриевой соли реагента класса (молекулярный вес 36,000-50,000 Da) </td><td> MP Biomedicals </td><td> 160110 </td><td></td></tr><tr><td> Gen5 (программное обеспечение) </td><td> Биотек </td><td> Версия 1.10.8 </td><td></td></tr><tr><td> Hexadecyltrimethylammonium бромид (HTAB) </td><td> Sigma-Aldrich </td><td> H5882-100G </td><td></td></tr><tr><td> Перекись водорода, 30 мас.% Раствора в воде </td><td> Sigma-Aldrich </td><td> 216763 </td><td> Хранить при температуре 2-8 ° C </td></tr><tr><td> Микротест плел 96-луночного плоским дном </td><td> Sarstedt </td><td> 82,1581 </td><td> Для однократного использования </td></tr><tr><td> -дианизидина </td><td> Sigma-Aldrich </td><td> D-3252 </td><td> Свет чувствительны. Хранить при температуре 2-8 ° C </td></tr><tr><td> Калий фосфат, двузамещенный </td><td> Каледон </td><td> 6620-1 </td><td></td></tr><tr><td> Калия фосфат однозамещенный </td><td> EMD Химические вещества </td><td> PX1565-1 </td><td></td></tr><tr><td> Ингибитор протеазы коктейль </td><td> Sigma-Aldrich </td><td> P8340 </td><td> Хранить при температуре от -20 ° C </td></tr><tr><td> Тритон Х-100 </td><td> Sigma-Aldrich </td><td> T8787 </td><td></td></tr><tr><td> Карбид вольфрама шарики для тканей Lyser II </td><td> QUIAGEN </td><td> 69997 </td><td></td></tr></tbody></table>

investigating intestinal inflammation in dss-induced model of ibd

Inflammatory bowel disease (IBD) encompasses a range of intestinal pathologies, the most common of which are ulcerative colitis (UC) and Crohn's Disease (CD). Both UC and CD, when present in the colon, generate a similar symptom profile which can include diarrhea, rectal bleeding, abdominal pain, and weight loss.1 Although the pathogenesis of IBD remains unknown, it is described as a multifactorial disease that involves both genetic and environmental components.2
There are numerous and variable animal models of colonic inflammation that resemble several features of IBD. Animal models of colitis range from those arising spontaneously in susceptible strains of certain species to those requiring administration of specific concentrations of colitis-inducing chemicals, such as dextran sulphate sodium (DSS). Chemical-induced models of gut inflammation are the most commonly used and best described models of IBD. Administration of DSS in drinking water produces acute or chronic colitis depending on the administration protocol.3 Animals given DSS exhibit weight loss and signs of loose stool or diarrhea, sometimes with evidence of rectal bleeding.4,5 Here, we describe the methods by which colitis development and the resulting inflammatory response can be characterized following administration of DSS. These methods include histological analysis of hematoxylin/eosin stained colon sections, measurement of pro-inflammatory cytokines, and determination of myeloperoxidase (MPO) activity, which can be used as a surrogate marker of inflammation.6
The extent of the inflammatory response in disease state can be assessed by the presence of clinical symptoms or by alteration in histology in mucosal tissue. Colonic histological damage is assessed by using a scoring system that considers loss of crypt architecture, inflammatory cell infiltration, muscle thickening, goblet cell depletion, and crypt abscess.7 Quantitatively, levels of pro-inflammatory cytokines with acute inflammatory properties, such as interleukin (IL)-1&beta;, IL-6 and tumour necrosis factor (TNF)-&alpha;,can be determined using conventional ELISA methods. In addition, MPO activity can be measured using a colorimetric assay and used as an index of inflammation.8
In experimental colitis, disease severity is often correlated with an increase in MPO activity and higher levels of pro-inflammatory cytokines. Colitis severity and inflammation-associated damage can be assessed by examining stool consistency and bleeding, in addition to assessing the histopathological state of the intestine using hematoxylin/eosin stained colonic tissue sections. Colonic tissue fragments can be used to determine MPO activity and cytokine production. Taken together, these measures can be used to evaluate the intestinal inflammatory response in animal models of experimental colitis.

Watch this Scientific Journal Video about Investigating Intestinal Inflammation in DSS-induced Model of IBD at JoVE.com

Investigating Intestinal Inflammation in DSS-induced Model of IBD

Экспериментальные модели воспалительных заболеваний кишечника позволили нам изучить сложный врожденный и адаптивный иммунный ответ связана с патогенезом. Использование гистологического забил, количественное определение провоспалительных цитокинов и активность миелопероксидазы, можно приступить к оценке этих ответов видно при воспалительных заболеваниях кишечника.

Исследование кишечного воспаления в DSS-индуцированной модели IBD

Inflammatory bowel disease (IBD) encompasses a range of intestinal pathologies, the most common of which are ulcerative colitis (UC) and Crohn's Disease ...

Research

JoVE Journal

Biology

Catheterization of Intestinal Loops in Ruminants

Катетеризация кишечных петель у жвачных

The intestine is a complex structure that is involved not only in absorption of nutrients, but also acts as a barrier between the individual and the ...

Rat Mesentery Exteriorization: A Model for Investigating the Cellular Dynamics Involved in Angiogenesis

Крысы Брыжейка экстериоризации: модель для исследования Клеточная динамика, вовлеченных в ангиогенез

Microvacular network growth and remodeling are critical aspects of wound healing, inflammation, diabetic retinopathy, tumor growth and other disease ...

Investigating the Spreading and Toxicity of Prion-like Proteins Using the Metazoan Model Organism C. elegans

Investigating the Spreading and Toxicity of Prion-like Proteins Using the Metazoan Model Organism C. elegans

Исследуя распространение и токсичности прионами как белков с использованием многоклеточных животных модельного организма<em&gt; C. Элеганс</em

Prions are unconventional self-propagating proteinaceous particles, devoid of any coding nucleic acid. These proteinaceous seeds serve as templates for ...

A Protocol for Lentiviral Transduction and Downstream Analysis of Intestinal Organoids

Протокол Lentiviral трансдукции и вниз по течению анализа заболеваемости кишечными органоиды

Intestinal crypt-villus structures termed organoids, can be kept in sustained culture three dimensionally when supplemented with the appropriate growth ...

A Mouse Model of Intestinal Partial Obstruction

Мыши модель частичной непроходимостью кишечника

Intestinal obstructions, that impede or block peristaltic movement, can be caused by abdominal adhesions and most gastrointestinal (GI) diseases including ...

Mass Spectrometry Analysis to Identify Ubiquitylation of EYFP-tagged CENP-A (EYFP-CENP-A)

Mass Spectrometry Analysis to Identify Ubiquitylation of EYFP-tagged CENP-A EYFP-CENP-A

Анализ масс-спектрометрии для выявления повсеместного ания метеоуожего тега CENP-A (EYFP-CENP-A)

Studying the structure and the dynamics of kinetochores and centromeres is important in understanding chromosomal instability (CIN) and cancer ...

Organoid-Derived Epithelial Monolayer: A Clinically Relevant In Vitro Model for Intestinal Barrier Function

Органоидный эпителиальный монослой: клинически значимая модель in vitro для кишечной барьерной функции

In the past, intestinal epithelial model systems were limited to transformed cell lines and primary tissue. These model systems have inherent limitations ...

Intestinal Epithelial Regeneration in Response to Ionizing Irradiation

Регенерация эпителия кишечника в ответ на ионизирующее облучение

The intestinal epithelium consists of a single layer of cells yet contains multiple types of terminally differentiated cells, which are generated by the ...

Epithelial Effects of Intestinal Inflammation on Established Murine Colonoids

Studying the Epithelial Effects of Intestinal Inflammation In Vitro on Established Murine Colonoids

Studying the Epithelial Effects of Intestinal Inflammation In Vitro on Established Murine Colonoids (Video) | JoVE 

Изучение эпителиальных эффектов воспаления кишечника in vitro на установленных колоноидах мышей

The intestinal epithelium plays an essential role in human health, providing a barrier between the host and the external environment. This highly dynamic ...

Construction of a 3D Intestinal Mucosa Model with Paraffin Embedding

Basic Three-Dimensional (3D) Intestinal Model System with an Immune Component

Investigating the Effects of Compounds on Intestinal Tissue Using 3D Human Cell Line Models (Video) | JoVE

Базовая трехмерная (3D) система моделей кишечника с иммунным компонентом

There has been an increase in the use of in vivo and in vitro intestinal models to study the pathophysiology of inflammatory intestinal diseases, for the ...