Name: intrarenal injection of escherichia coli in a rat model of pyelonephritis
Uploaded: 2017-07-18T12:30:00
Description: Watch this Scientific Journal Video about Intrarenal Injection of Escherichia coli in a Rat Model of Pyelonephritis at JoVE.com

The authors would like to acknowledge the support of NIH/NIDDK K12 DK100014 (Lan Lu), the Case Comprehensive Cancer Center (NIH/NCI P30 CA43703), and the Clinical and Translation Science Collaborative of Cleveland (NIH/NCATS UL1 TR000439).

Alle dyreforsøk ble utført i henhold til godkjente protokoller for institusjonell dyrepleie og brukskomité (IACUC) ved Case Western Reserve University. Varigheten av den kirurgiske prosedyren beskrevet nedenfor er ca. 45-60 minutter. MR-prosedyren i seg selv er ca. 15 minutter for hvert tidspunkt. 1. Anestesi <ol><li> Bedøv råtten i isoflurankammer satt til 2% isofluran blandet med oksygen for å lette dyrehåndtering og fastholdelse før administrering av injiserbar anestesi intraper...

<ol>
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Stigende akutt pyelonefrit hos gnagere (dvs. mus og rotter) kan produseres ved transuretral kateterisering. 16 , 17 , 18 Denne transuretrale infeksjonsmetoden er fordelaktig ved at den er ikke-invasiv og etterligner den menneskelige patofysiologi av stigende infeksjon. 17 , 18 , 19 , 20 Imidl...

Gnagere modeller har blitt brukt til å studere mange menneskelige sykdoms manifestasjoner, inkludert pyelonefrit og urinveisinfeksjoner (UTI). UTI er et globalt helseproblem, og kan påvirke barn, menn og kvinner i alle aldre. 1 , 2 , 3 Den første manifestasjonen av UTI-er inkluderer blærebetennelse, og hvis infeksjonen stiger opp langs urineren, kan en nyreinfeksjon (pyelonefrit) følge. Samtidig nærmer utbredelsen av diabetes 400 millioner mennesker over hele verden. 4 , 5 Det er viktig at UTI-forekomsten kan være opptil 4 ganger høyere hos pasienter som er overvektige eller har type 2 diabetes mellitus, noe som resulterer i økt risiko for gjentatt UTI-infeksjon (RUTI), sepsis, nyrefibrose fra pyelonefrit og blæredysfunksjon. 6 , 7 , 8 GnagereModeller er viktige for å studere UTI, fordi nåværende antibiotikabehandlinger produserer en vedvarende, forebyggende respons bare i en delmengde av UTI-pasienter. For å forbedre klinisk UTI-omsorg er hovedtrinnene å forstå mekanismen for RUTI og dens patofysiologiske prosesser fra akutt infeksjon til betennelse i fibrose, samt virkningen av type 2 diabetes mellitus. Målet med å forbedre dyremodeller er å utvikle teknikker som muliggjør en mer nøyaktig vurdering av sykdomsprogresjon og terapeutiske inngrep. Flere forskjellige tilnærminger har blitt anvendt for å indusere pyelonefrit hos rotter og / eller mus for å studere patofysiologien av nyrebeskadigelse, effekten av antibiotisk behandling og andre aspekter av den naturlige forlengelsen av UTI. En felles tilnærming til å etablere retrograd UTI er transuretral kateterisering. 10 , 11 , 12 , 13 &lt;/ Sup&gt; Denne metoden introduserer bakterier via urinrøret i urinblæren hos bedøvede dyr. Selv om denne teknikken nøye simulerer menneskelig pyelonefrit, kan den faktiske forekomsten og størrelsen på pyelonefritisinfeksjonen være svært variabel på grunn av flere faktorer, inkludert mangel på spontan ureterisk refluks eller urinrengjøring under eller umiddelbart etter inokulering. 11 Som et resultat av eksperimentell variabilitet i å indusere en stigende pyelonefritt infeksjon kan begrense anvendeligheten av denne modell for å studere nyre-infeksjoner, så vel som terapeutiske strategier. Denne rapporten beskriver en kirurgisk pyelonephritis rotte modell hvor E. coli injiseres direkte i rotteryren. Til tross for at denne rottemodellen er invasiv, kan mengden av E. coli levert til nyrene effektivt kontrolleres slik at det blir en robust nyreinfeksjon og betennelse. 14 Innenfor denne prosedyren beskriver vi ogsåHvordan disse induserte nyreinfeksjonene kan overvåkes langsgående med in vivo magnetisk resonansimaging (MR).

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			<td height="21" style="height:21px;width:216px;">E. coli&#160; UTI89 or CFT073</td>
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intrarenal injection of escherichia coli in a rat model of pyelonephritis

Pyelonephritis is a bacterial infection of the kidney and is most commonly caused by Escherichia coli. Recurrent infections can cause significant renal inflammation and fibrosis ultimately resulting in declining kidney function. Before improved clinical management and prevention of pyelonephritis can be instituted, a reliable animal model must be established in order to study the mechanisms of progression, recurrence, and therapeutic efficacy. The transurethral infection model closely mimics human pyelonephritis but exhibits considerable variation due to its reliance on urethral reflux to transport the bacteria to the kidney. Herein, a detailed surgical protocol for performing bacterial injections into the rat renal pelvis is provided and confirmed by non-invasive Magnetic Resonance Imaging (MRI). Using this protocol, animals receive direct exposure to a desired concentration of E. coli bacteria and can fully recover from the surgical procedure with adequate post-operative care. This facilitates subsequent longitudinal MRI assessments of the experimental animal models for comparison with saline (sham) controls. Using this direct delivery approach, the severity of infection is controllable and applicable for mechanistic studies of progression as well as development of novel treatment strategies.

Medisinsk billedteknikk gir mulighet til å vurdere ikke-invasiv UTI og terapeutisk effekt. Derfor MRI ble anvendt for å validere induksjon av akutt infeksjon etter injeksjon av 1-2 x 10 7 UTI89 E. coli, og for å visualisere endringene i nyrene før og etter operasjonen. Figur 1a-b viser et gradvis økende område av nyreinfeksjon (gule piler). MR-bilder oppnådd for hvert dyr i dag 1 og 4 etter infeksjon, bidrar til å karakterisere veksten av den ...

Watch this Scientific Journal Video about Intrarenal Injection of Escherichia coli in a Rat Model of Pyelonephritis at JoVE.com

Intrarenal Injection of Escherichia coli in a Rat Model of Pyelonephritis

This manuscript describes a rat surgical model of pyelonephritis using direct intra-renal infection by Escherichia coli into the renal pelvis. The experimental procedure can be utilized to study the pathogenesis of pyelonephritis as well as the associated inflammation and fibrosis.

Intrarenal injeksjon av<em&gt; Escherichia coli</em&gt; I en rotte modell av pyelonefritis

Pyelonephritis is a bacterial infection of the kidney and is most commonly caused by Escherichia coli. Recurrent infections can cause significant renal ...

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Intrarenal Injection of Escherichia coli in a Rat Model of Pyelonephritis

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