葡聚糖硫酸钠（DSS）诱导的结肠炎小鼠模型

Srustidhar Das; Surinder K. Batra; Satyanarayana Rachagani

doi:10.21769/BioProtoc.2515

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Peer-reviewed

Mouse Model of Dextran Sodium Sulfate (DSS)-induced Colitis

葡聚糖硫酸钠（DSS）诱导的结肠炎小鼠模型

Srustidhar Das

SB Surinder K. Batra

SR Satyanarayana Rachagani email

发布: 2017年08月20日第7卷第16期 DOI: 10.21769/BioProtoc.2515 浏览次数: 18810

评审: Pasquale PellegriniAnonymous reviewer(s)

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Cover of Oncogene, featuring study using the protocol.

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Abstract

Inflammatory bowel disease (IBD) is a chronic inflammatory disease of the intestinal tract and is primarily comprised of Crohn’s disease (CD) and ulcerative colitis (UC). Several murine models that include both chemical induced and genetic models have been developed that mimic some aspects of either CD or UC. These models have been instrumental in our understanding of IBD. Of the chemical induced colitis models, dextran sodium sulfate (DSS) induced colitis model is a relatively simple and very widely used model of experimental colitis.

Keywords: Dextran sodium sulfate (DSS) (葡聚糖硫酸钠（DSS）)

Colitis (结肠炎)

Inflammatory bowel disease (IBD) (炎性肠病（IBD）)

Background

Inflammatory bowel disease (IBD) is a complex and multifactorial disease of unknown etiology (Sartor, 2006). However, multiple factors are considered to be critical in conferring IBD susceptibility, e.g., defects in host genetics, environmental triggers, aberrant immune response against microbial and dietary antigens (Sartor, 2006). Several murine models that address specific aspects of the disease process are available (Mizoguchi, 2012). DSS induced experimental colitis is a rapid and widely used model of intestinal inflammation (Perse and Cerar, 2012). Although the exact mechanism of DSS induced colitis is not well understood, it is widely accepted that disruption of the epithelial monolayer resulting in exposure of the underlying immune system to the intestinal contents containing microbiota and microbial products (Perse and Cerar, 2012). DSS induced colonic inflammation can be adapted as acute, chronic or relapsing model of intestinal inflammation by changing the concentration, duration and cycles of administration of DSS in drinking water. In this protocol, we provide a detailed description of procedures, important considerations when performing the protocol. We have used this protocol to address the role of Muc4 in DSS induced colitis in our previous work (Das et al., 2015).

Materials and Reagents

Sterile Eppendorf tubes (National Scientific, catalog number: CN1700-BP )
Tissue Path Macrosette processing cassettes (Fisher Scientific, catalog number: 15-182-706 )
Glass slides
8-10 week old mice generated in house or obtained from the commercial vendors
Note: Mice used in the study of Das et al., (2015) were of 129/Sv and C57BL/6J mixed background. Mice in this study were littermates and included both males and females housed under specific pathogen free (SPF) conditions.
Dextran sodium sulfate (DSS) (TdB Consultancy, catalog number: DB001 , molecular weight: 35-55 kDa)
Note: Stored in a dry place at room temperature.
Liquid nitrogen
Autoclaved drinking water
Anesthesia (Isofluorane or CO₂)
10% buffered formalin (Fisher Scientific, catalog number: SF100-4 )
Ethanol
Xylene
Paraffin
mirVana miRNA isolation kit (Thermo Fisher Scientific, Invitrogen^TM, catalog number: AM1560 )
Bioanalyzer (Agilent Technologies, Waldbronn, Germany)
DNase I (QIAGEN, catalog number: 79254 )
Oligo-dT or random hexamer primers (Thermo Fisher Scientific, Invitrogen^TM, catalog number: 100023441 )
SuperScript reverse transcriptase II (Thermo Fisher Scientific, Invitrogen^TM, catalog number: 18064014 )
Light Cycler 480 SYBR Green mix (Roche Molecular Systems, catalog number: 04707516001 )
Hematoxylin and eosin (H & E)

Equipment

Measuring scale/Ruler or Vernier calipers
Animal weighing balance
Feeding bottles
Tissue-Tek VIP processing machine
Dissection equipment
Mouse microisolator chambers
Pestle and mortar

Procedure

English

中文翻译

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如何引用

Das, S., Batra, S. K. and Rachagani, S. (2017). Mouse Model of Dextran Sodium Sulfate (DSS)-induced Colitis. Bio-protocol 7(16): e2515. DOI: 10.21769/BioProtoc.2515.