Name: murine aortic crush injury: an efficient in vivo model of smooth muscle cell proliferation and endothelial function
Uploaded: 2017-06-11T14:00:00
Description: Watch this Scientific Journal Video about Murine Aortic Crush Injury: An Efficient In Vivo Model of Smooth Muscle Cell Proliferation and Endothelial Function at JoVE.com

Vi takker Hsia Ru-ching PhD, fra Electron Microscopy Core Facility ved University of Maryland School of Medicine, for hennes tekniske støtte i behandlingen av scannings elektroniske mikroskopi prøver.

Etikkerklæring: Protokollene for dyrehåndtering ble godkjent av Institutt for dyrepleie og brukskomité (IACUC) ved University of Maryland (protokollnummer 0416009) og utført i henhold til AAALAC-internasjonale standarder. 1. Kirurgisk Prosedyre <ol><li> Anestetisk teknikk <ol><li> Steriliser alle instrumenter som brukes i overlevelsekirurgi med dampsterilisering ved 121 ° C i 30 minutter. </li><li> Induktor anestesi via en induksjonstank med 100% O2 og 2,5% i...

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Vi har karakterisert effekten av en muse-aortisk skade modell som resulterer i medial hyperplasi og endotel barriere dysfunksjon. Delvis EC-frigjøring langs aorta-intima ledsaget av tap av cellecellekontakt og forbedring av cellefremspring. Tilsvarende var endotelbarrierefunksjonen signifikant svekket, noe som stimulerte de mitogenfølsomme signalveiene, noe som førte til proliferasjon av VSMC og fortykkelse av karvegveggen. Styrken i denne modellen er at det er teknisk lettere å lære og utføre enn andre aorta-mode...

restenose Følgende kardiovaskulære prosedyrer (bypass kirurgi, angioplastikk eller stenting) er et betydelig problem som reduserer holdbarheten av disse prosedyrene. Alle revaskulariseringsprosedyrer plages av restenose. Nåværende strategier for å hindre restenose (legemiddeleluerende stenter og medikamentbelagte ballonger) hemmer både vaskulær glattmuskelcelle (VSMC) og endotelcelleproliferasjon (EC). Følgelig forhindrer disse intervensjonene VSMC-mediert restose, men forhindrer også regenerering av endotelet. Uten intakt endotel skal pasienter være på potente antiplatelet midler for å redusere risikoen for in situ trombose med risiko for blødningskomplikasjoner. En ideell terapi vil hemme VSMC-proliferasjon samtidig som det fremmer regenerering av endotelet. Dermed er det et behov for samtidig å studere VSMC proliferasjon og endotelbarrierefunksjon i n vivo . For tiden er det alvorligAl musemodeller av restenose 1 . Disse modellene inkluderer carotidligasjon og femoralt arterie ledningsskade 2 . Aorta modeller inkluderer stent plassering 3 , ballong skade 4 og aorta allograft 5 . Alle de nåværende modellene er begrenset. Carotidligering genererer en strømningsmediert neointimal lesjon og har ikke endotelskader. I tillegg har begge karotid- og femorale arterier mange ganger færre cellelag enn menneskeskap, og begrenser deres translasjonsverdi. Musaortaen som er ca. 1,3 mm i diameter, er det eneste fartøyet som tilnærmer seg en klinisk relevant (kransartet) menneskearterie (3). Til tross for det translasjonelle potensialet for muse-aorta-modeller av sykdom, har nåværende modeller begrensninger. Disse modellene krever avanserte mikrokirurgiske ferdigheter og spesialisert utstyr som angioplastikkballonger og stenter. Her presenterer viEn roman, reproduserbar teknikk for samtidig å indusere VSMC proliferasjon og forstyrre endotelbarrierefunksjonen.

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murine aortic crush injury: an efficient in vivo model of smooth muscle cell proliferation and endothelial function

Arteriell rekonstruksjon, enten angioplastikk eller bypass-kirurgi, involverer iatrogen trauma som forårsaker endotelavbrudd og vascular glatt muskelcelle (VSMC) proliferasjon. Vanlige murine modeller studerer små fartøy som karoten og femorale arterier. Her beskriver vi et in vivo system hvor både VSMC proliferasjon og endotelbarrierefunksjonen kan vurderes samtidig i en stor beholder. Vi studerte infrarenal aorta respons på skade i C57BL / 6 mus. Aorta ble skadet fra venstre renalvein til aorta-bifurkasjon med 30 transmurale knusninger med 5 sekunder lang varighet med en bomullstipsapplikator. Morfologiske endringer ble vurdert med konvensjonell histologi. Aorta-veggtykkelsen ble målt fra luminalflaten til adventitia. EdU-integrasjon og tellerfarvning med DAPI og alpha-actin ble brukt til å demonstrere VSMC-proliferasjon. Aktivering av ERK1 / 2, en kjent moderator for intimal hyperplasi-dannelse, ble avskrekketUtvunnet av Western Blot-analyse. Effekten av betennelse ble bestemt ved immunhistokjemi for B-celler, T-celler og makrofager . En ansiktsseksjoner av endotelet ble visualisert med skanningelektronmikroskopi (SEM). Endotelbarrierefunksjonen ble bestemt med Evans Blue-farging. Transmural skade resulterte i aorta veggtykkelse. Denne skaden induserte VSMC proliferasjon, mest fremtredende ved 3 dager etter skade, og tidlig aktivering av ERK1 / 2 og redusert p27 kip1 ekspression. Skade resulterte ikke i økte B-celler, T-celler eller makrofager infiltrering i kargen veggen. Skade forårsaket delvis endotelcelle denudasjon og tap av cellecellekontakt. Skade resulterte i et signifikant tap av endotelbarrierefunksjonen, som returnerte til baseline etter syv dager. Den murine transmurale slanke aorta-skademodellen gir et effektivt system for samtidig å studere både VSMC-proliferasjon og endotelbarrierefunksjon i en stor beholder.

Tverrsnitt aorta innebygd i OCT ble snittet og farget med hematoksylin og eosin, og deretter tellert farget med Verhoeff-Van Gieson (VVG) flekk for å identifisere det indre og ytre elastiske laminat 7 . Kryssskade forårsaket aorta-veggtykkelse sammenlignet med aortaene av dyr behandlet med en sham-prosedyre (laparotomi og liten tarmmobilisering alene). Veggtykkelse, som vurderes av avstanden fra adventitia til lumen, var størst tre dager etter skade (42,2 ± 1,...

Watch this Scientific Journal Video about Murine Aortic Crush Injury: An Efficient In Vivo Model of Smooth Muscle Cell Proliferation and Endothelial Function at JoVE.com

Murine Aortic Crush Injury: An Efficient In Vivo Model of Smooth Muscle Cell Proliferation and Endothelial Function

Restenose etter kardiovaskulære prosedyrer (bypass kirurgi, angioplastikk eller stenting) er et betydelig problem som reduserer holdbarheten av disse prosedyrene. En ideell terapi vil hemme proliferasjon av glatt muskelcelle mens den fremmer regenerering av endotelet. Vi beskriver en modell for samtidig vurdering av VSMC proliferasjon og endotelfunksjon in vivo.

Murine Aortic Crush Skade: En Effektiv<em&gt; In vivo</em&gt; Modell av jevn spredning av muskelceller og endotelfunksjon

Arterial reconstruction, whether angioplasty or bypass surgery, involves iatrogenic trauma causing endothelial disruption and vascular smooth muscle cell ...

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Murine Aortic Crush Injury: An Efficient In Vivo Model of Smooth Muscle Cell Proliferation and Endothelial Function

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