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

Dette arbejde er støttet af tilskud fra Canadian Institutes of Health Research (CIHR) og Crohns og Colitis Foundation of Canada (CCFC).

1. Murine model af DSS-induceret akut colitis <ol><li> Tilføj dextransulfat natrium (DSS) til autoklaveret drikkevand til den ønskede slutkoncentration (1-5%) (wt / vol) (dvs. at gøre en 5% DSS løsning, tilsættes 25 mg af DSS pulver til 500 ml autoklaveret vand) . Stamopløsning kan stå ved stuetemperatur i op til en uge eller ved 4 ° C indtil brug. </li><li> I et biosikkerhed hætte, hæld lager DSS opløsning i 50 ml Falcon rør (en nødvendig pr bur). Hold stamopløsning at genopfylde rør når det er nødve...

<ol>
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DSS colitis er en meget anvendt kemisk induceret model af tarmbetændelse. I denne model er mus, der fik drikkevand suppleret med DSS, som menes at være giftige for tarm epitel celler og forstyrre integritet mucosal barriere. 10 Administration af DSS inducerer et akut colitis, der er karakteriseret ved løs afføring, fækal blødning, og infiltration med granulocytter. 10 Under DSS administration er colitis normalt forbundet med signifikant vægttab og tilstedeværelse af blod i afføringen, hvil...

<table border="1"><tbody><tr><td> Navn på reagens / udstyr: </td><td> Firma </td><td> Katalog nummer </td><td> Kommentarer </td></tr><tr><td> Eppendorf Safe-Lock mikrocentrifugerør (2 ml) </td><td> Eppendorf </td><td> 0030 120.094 </td><td></td></tr><tr><td> BIOTEK EL808 Absorbans pladelæser </td><td> BIOTEK </td><td> EL808 </td><td></td></tr><tr><td> Dextransulfat natriumsalt reagens bedømmelse (molekylvægt 36,000-50,000 Da) </td><td> MP Biomedicals </td><td> 160110 </td><td></td></tr><tr><td> Gen5 (software) </td><td> BIOTEK </td><td> Version 1.10.8 </td><td></td></tr><tr><td> Hexadecyltrimethylammonium bromid (HTAB) </td><td> Sigma-Aldrich </td><td> H5882-100G </td><td></td></tr><tr><td> Hydrogenperoxid, 30 wt.% Opløsning i vand </td><td> Sigma-Aldrich </td><td> 216763 </td><td> Opbevares ved 2-8 ° C </td></tr><tr><td> MICROTEST plspiste 96-såvel flad bund </td><td> Sarstedt </td><td> 82,1581 </td><td> Kun til engangsbrug </td></tr><tr><td> o-Dianisidin </td><td> Sigma-Aldrich </td><td> D-3252 </td><td> Lysfølsomme. Opbevares ved 2-8 ° C </td></tr><tr><td> Kalium, dibasisk </td><td> Caledon </td><td> 6620-1 </td><td></td></tr><tr><td> Kalium, monobasisk </td><td> EMD Kemikalier </td><td> PX1565-1 </td><td></td></tr><tr><td> Proteasehæmmer cocktail </td><td> Sigma-Aldrich </td><td> P8340 </td><td> Opbevares ved -20 ° C </td></tr><tr><td> Triton X-100 </td><td> Sigma-Aldrich </td><td> T8787 </td><td></td></tr><tr><td> Wolfram Carbide perler for Tissue Lyser II </td><td> QUIAGEN </td><td> 69997 </td><td></td></tr></tbody></table>

investigating intestinal inflammation in dss-induced model of ibd

Inflammatorisk tarmsygdom (IBD) omfatter en række af tarm sygdomme, de mest almindelige af dem er colitis ulcerosa (UC) og Crohns sygdom (CD). Både UC og CD, når de findes i tyktarmen, generere en lignende symptom profil, der kan omfatte diarré, blødning fra endetarmen, mavesmerter og vægttab. 1 Selv om patogenesen af IBD er fortsat ukendt, er det beskrevet som en multifaktoriel sygdom, der involverer både genetiske og miljømæssige komponenter. 2 Der er mange og varierende dyremodeller af colon inflammation, der ligner flere funktioner af IBD. Dyremodeller af colitis spænder fra dem, der opstår spontant i følsomme stammer af visse arter til dem, der kræver administration af bestemte fusioner af colitis-inducerende kemikalier, som f.eks dextransulfat natrium (DSS). Kemisk-inducerede modeller af tarmen inflammation er den mest udbredte og bedst beskrives modeller af IBD. Adminidemonstration af DSS i drikkevandet producerer akut eller kronisk colitis, afhængigt af administrationen protokollen. 3 dyr, der fik DSS udviser vægttab og tegn på tynd afføring eller diarré, undertiden med tegn på blødning fra endetarmen. 4,5 Her beskriver vi de metoder, hvormed colitis udvikling og den deraf følgende inflammatoriske respons kan karakteriseres efter administration af DSS. Disse metoder omfatter histologisk analyse af hæmatoxylin / eosin farves kolon sektioner, måling af pro-inflammatoriske cytokiner, og bestemmelse af myeloperoxidase (MPO) aktivitet, som kan bruges som et surrogat markør for inflammation. 6 Omfanget af den inflammatoriske reaktion i sygdommen tilstand kan vurderes ved tilstedeværelse af kliniske symptomer eller ved ændring i histologi i slimhindevævet. Colon histologiske skader vurderes ved hjælp af et pointsystem, der betragter tab af krypt arkitektur, inflammatoriske celleinfiltration, muskel thickening, bæger celle depletion og krypt byld. 7 Kvantitativt kan niveauer af pro-inflammatoriske cytokiner med akut inflammatoriske egenskaber, såsom interleukin (IL)-1β, IL-6 og tumornekrosefaktor (TNF)-α, bestemmes ved hjælp af konventionelle ELISA-metoder. Desuden kan MPO aktivitet måles ved hjælp af en kolorimetrisk analyse og bruges som et indeks af inflammation. 8 I eksperimentel colitis, er sygdommens sværhedsgrad ofte er korreleret med en stigning i MPO aktivitet og højere niveauer af pro-inflammatoriske cytokiner. Colitis sværhedsgrad og betændelse-relaterede skader kan vurderes ved at undersøge afføring konsekvens og blødning, udover at vurdere den histopatologiske tilstand af tarmen ved hjælp af hæmatoxylin / eosin farves colonic vævssnit. Colon væv fragmenter kan bruges til at bestemme MPO aktivitet og cytokin produktion. Tilsammen kan disse foranstaltninger anvendes til at vurdere den intestinale inflammatorisk respons idyremodeller for eksperimentel 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

Eksperimentelle modeller af inflammatorisk tarmsygdom har tilladt os at undersøge det komplekse medfødte og adaptive immunrespons i forbindelse med patogenese. Ved hjælp af histologiske scoring, kvantificering af pro-inflammatoriske cytokiner og myeloperoxidase aktivitet, kan man begynde at vurdere disse svar ses i inflammatorisk tarmsygdom.

Undersøgelse tarmbetændelse i DSS-induceret Model af IBD

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

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