Name: ileectomy-induced bile overaccumulation in mouse intestine
Uploaded: 2017-08-21T13:00:00
Description: Watch this Scientific Journal Video about Ileectomy-induced Bile Overaccumulation in Mouse Intestine at JoVE.com

Denne forskning er understøttet af Tom Peterson Foundation og NIH give R01-HL119780 (Jain, MK).

animalske protokol blev gennemgået og godkendt af institutionelle Animal Care og brug Udvalget ved Case Western Reserve University School of Medicine og blev gennemført i overensstemmelse med National Institutes of Health (NIH) Guide for den Pleje og brugen af forsøgsdyr (8. udgave, 2011). Mus blev aflivet efter metoder, der er kompatibel med de amerikanske veterinære Medical Association (AVMA) retningslinjer for aflivning af dyr (2013 udgave). C57BL/6J, mandlige, 8-16 uge gamle mus blev brugt i denne protokol. Mus ...

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For at udføre en vellykket ileectomy, skal den overlegne mesenteriallymfeknuderne arterie være forbundet i forvejen for at blokere blodforsyningen til det resecting segment. Den iskæmisk ileal segment vil vende mørk lilla efter ligatur. Den ileal segment skal derefter være fuldstændig resektion. En normal blodforsyning skal sikres for de opbevarede ender. Dette er afgørende for at undgå blødning og ukorrekt fjernelse, der kan let forårsage kirurgiske fejl på grund af iskæmisk nekrose efter suturering. Under a...

I moderne biomedicinsk forskning udnyttes genetisk manipulerede dyremodeller bredt for at opsnuse indsigt i menneskelige sygdomme. Især har væv eller celle-specifikke funktioner, gevinst og tab af gener været brugt til at studere molekylære forordning samt induceret biologiske virkninger. Trods fremskridt i manipulere target gener i vivo, er der dvælende begrænsninger. Første, mange celler eller væv specifikke sletninger påvirker flere organer. For eksempel, vil epithelial gen sletning fjerne udtryk i epitheler i flere væv. Yderligere, selv om sletning er begrænset til en bestemt væv, fysisk kontrol er sjældent muligt. For eksempel, i et væv lignende tarmen udføre adskilte segmenter meget specifikke funktioner, der ikke kan manipuleres med præcision i vivo. I disse situationer er resektion af gen-holdige væv anses for at være en mere effektiv studere tilgang til at bestemme den mekanistiske og funktionel betydning af væv meddelelse.
Ileectomy er for det meste brugt i patienter med Crohns og inflammatoriske sygdomme der involverer distale ileum 1,2,3. Ileum typisk producerer flere energy storage hormoner som fibroblast vækstfaktor 15/19 (FGF15/19), peptid YY (PYY) og glukagon-lignende peptid 1/2 (GLP1/2); disse hormoner spiller en vigtig lokal og endokrine rolle i mange biologiske funktioner4,5,6. Blandt disse hormoner, er FGF15 blevet identificeret som en robust endokrine hæmmer af galdesyre syntese i leveren. Når reabsorberes i ileal enterocytes, aktivere galdesyrer nukleare receptor farnesoid X receptor (FXR) til at stimulere Fgf15 udtryk, der efterfølgende fører til feedback hæmning af hepatisk galdesyre syntese 7. I en nylig undersøgelse, vi indførte mus ileectomy model for at studere den ileal kruppel-lignende faktor 15 (KLF15) -Fgf15 signalering akse, der regulerer døgnrytmen galdesyre produktion i leveren 8. Vigtigst, indført vi en roman familie, kruppel-lignende faktorer, navnlig KLF15, i galdesyre biologi. Baseret på funktionelle studier herunder ileectomy kirurgi, vi har besluttet at KLF15 upregulates galdesyre syntese via en indirekte mekanisme til ikke-hepatisk. Endelig ileal KLF15 også er identificeret som den første endogene negativ regulator af Fgf15.
De intestinale segmenter faldende fra proksimalt for distale regioner er ansvarlige for absorption af forskellige næringsstoffer. Ileum er det største segment ansvarlig for galdesyre og vitamin B12 (VB12) absorption 9. En tidligere undersøgelse ansat en musemodel af proksimale tarm resektion at studere kort tyktarm; forskellige resektion længder, kostvaner og sutur typer blev foreslået til at opretholde en optimal efter operationen overlevelse af 10. En nyere gennemgang angiver desuden, at ileal resektion typisk resulterer i mere alvorlig sygdom end andre gastrointestinale (GI) segment resektion på grund af nedsat adaptive kapacitet af de resterende tarmkanal 11. Dette emne har fået intensiv interesser af grundforskning og klinisk forskningsgrupper, hvorimod forståelse af recovery og de effektive terapeutiske tilgange er stadig begrænset.
Galdesyre diarré skyldes ubalancer i galdesyre homøostase i enterohepatiske cirkulation 12,13. Det kan være en konsekvens af ileal resektion, gastrointestinal sygdom, eller et resultat af idiopatisk galdesyre malabsorption. Mere end 80% af patienterne har fundet for at præsentere med diarré efter undergår ileal resektion 14. Ileectomy har potentiale til at blive en vigtig kirurgi model for undersøgelse af galdesyre diarré. I denne undersøgelse, en række ileal resektion giver en gradient vurdering af FGF15 mangel samt intestinal galde salt malabsorption, overaccumulation og giftige skader.

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ileectomy-induced bile overaccumulation in mouse intestine

Tarm resektion er en fælles terapeutisk tilgang til menneskelige sygdomme som fedme, inflammatorisk tarmsygdom, Crohns sygdom, og tyktarmskræft, der ofte resulterer i alvorlige korte tarm syndrom-lignende bivirkninger herunder galdesyre diarré, dehydrering, elektrolyt forstyrrelser og næringsstof malabsorption. Her har vi indføre en murine ileal resektion model, såkaldte ileectomy, for at evaluere væv meddelelse og vedligeholdelse af systemisk homøostase. Efter ileal resektion er cirkulerende blod permanent ileum-specifikke endokrine hormon fibroblast vækst faktor 15 (FGF15), som frigiver sin endokrine hæmning af galdesyre syntese i leveren. I kombination med øget produktion og afskaffet reabsorption af galdesyrer efter fjernelse af ileum, lider mus, der blev opereret af galde salt overaccumulation i tarmen og tilknyttede diarré, sygelighed og dødelighed. Roman brug af kirurgi model indført i denne undersøgelse kan give mekanistiske og funktionelle indsigt i ileal kontrol af systemisk metabolisk regulering i fysiologi og sygdom.

Procedurerne for ileectomy er vist i figur 1. Det første trin omfatter forberedelse mus abdominal hud, gør en abdominal snit og bruge retraktorer til fuldt udsætte tarmen (figur 1A-C). Næste, mus coecum var placeret (fig. 1D); betragtning af, at dens størrelse og form gør det et let identificerbare vartegn. Coecum, ileum og en del af den distale jejunum blev ud...

Watch this Scientific Journal Video about Ileectomy-induced Bile Overaccumulation in Mouse Intestine at JoVE.com

Ileectomy-induced Bile Overaccumulation in Mouse Intestine

Tyndtarmen-afhængige galdesyre reabsorption og feedback hæmning af hepatisk galdesyre syntese er vigtigt for systemisk homøostase og sundhed. I denne undersøgelse beskriver vi en musemodel for ileal resektion at evaluere ileectomy-induceret galde malabsorption, overaccumulation og toksicitet i mus tarmen.

Ileectomy-induceret galde Overaccumulation i mus tarmen

Intestinal resection is a common therapeutic approach for human diseases such as obesity, inflammatory bowel disease, Crohn's disease, and colon cancer ...

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Intrauterin Telemetri måler Mouse kontraktile Pressure<em&gt; In Vivo</em

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. ...

A Simple Device to Rapidly Prepare Whole Mounts of the Mouse Intestine

En simpel anordning til hurtigt Forbered Hele Monteres i musetarm

Preparing whole mounts of the mouse small intestine and colon for subsequent analysis or quantification can be time consuming and difficult. We describe ...

Isolation and Profiling of MicroRNA-containing Exosomes from Human Bile

Isolering og Profilering af MicroRNA-holdige Exosomer fra human Bile

Exosome research in the last three years has greatly extended the scope towards identification and characterization of biomarkers and their therapeutic ...