Name: a mice model of chlorhexidine gluconate-induced peritoneal damage
Uploaded: 2022-04-28T14:45:00
Description: Watch this Scientific Journal Video about A Mice Model of Chlorhexidine Gluconate-Induced Peritoneal Damage at JoVE.com

Vi takker oprigtigt Shin-Han Tseng for den kritiske diskussion og delvise udførelse af undersøgelsen. Denne undersøgelse blev støttet af EDAHP110003 og NCKUEDA110002 fra Research Foundation of E-DA Hospital og National Cheng Kung University, Taiwan.

Alle museforsøg blev godkendt af Laboratory Animal Center på E-DA Hospital / I-Shou University og blev håndteret i henhold til "Guide for the Care and Use of Laboratory Animals" (NRC, USA 2011). C57BL/6-hanmus, 7-8 uger gamle, blev anvendt til nærværende undersøgelse.
1. Kemisk forberedelse
<ol><li>Forbered det kemiske irritationsmiddel ved at fortynde 0,1% chlorhexidingluconat (CG, se materialetabel) i 15% ethanol.</li>
</ol><str...

<ol>
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I denne undersøgelse præsenteres en mus PD-model ved i.p. injektion af CG, og resultaterne viste peritoneal fibrose og funktionel forringelse i denne model, som efterlignede PD-patientens tilstand.
Der er flere kritiske trin i protokollen. For det første, for at udføre en i.p. injektion af CG eller NS, skal musens abdominalvægshud afhentes ved hjælp af tang for at forhindre punkteringsinduceret intraperitoneal organskade. For det andet skal det område, der er beskadiget af ip-injektione...

Peritonealdialyse (PD) er en type nyreerstatningsterapi. PD har dog problemer, der ikke kan løses. For eksempel kan langvarig PD-behandling forårsage peritoneal skade, hvilket i sidste ende fører til ultrafiltreringssvigt og tilbagetrækning af behandling 1,2,3,4,5,6. Peritoneal fibrose er en af de mest alvorlige komplikationer 7,8. Peritoneal fibrose er karakteriseret ved aflejring og akkumulering af ekstracellulær matrix inden for interstitium og neo-angiogenese og vaskulopati i peritoneum 9,10.
Hovedårsagerne til disse peritoneale ændringer er tilbagevendende peritonitis og ikke-biokompatibilitet af dialysatet, som er hyperosmotisk, høj glukose, lav pH og glukosenedbrydningsproduktakkumulering11,12. Derfor kan egnede dyreforsøgsmodeller hjælpe forskere med bedre at studere bughindens fysiologiske og patologiske ændringer under PD-terapi. Derfor er etablering af en dyre-PD-model vigtig for at forbedre PD-teknologi og dialyseeffektivitet og forlænge patientens overlevelse. Denne undersøgelse havde til formål at generere en PD-musemodel ved intraperitoneal (i.p.) injektion af chlorhexidingluconat (CG), som beskrevet tidligere13,14. Denne PD-musemodel er enkel, nem at bruge og gennemførlig sammenlignet med andre PD-dyremodeller.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.9% Normal Saline</td>
			<td>Y F CHEMICAL CORP., New Taipei City, Taiwan</td>
			<td>-</td>
			<td></td>
		</tr>
		<tr>
			<td>10% neutral buffered formalin</td>
			<td>Taiwan Burnett International Co., Ltd., Taipei City, Taiwan</td>
			<td>00002A</td>
			<td></td>
		</tr>
		<tr>
			<td>Automatic biochemical analyzer</td>
			<td>Hitachi Ltd., Tokyo, Japan</td>
			<td>Labospect Series 008</td>
			<td>for determining glucose concentration</td>
		</tr>
		<tr>
			<td>Chlorhexidine digluconate solution, 20% in H2O</td>
			<td>Sigma-Aldrich, MO, USA</td>
			<td>C9394</td>
			<td>diluted to 0.1% with 15% ethanol for injection</td>
		</tr>
		<tr>
			<td>Ethanol</td>
			<td>Avantor Performance Materials, LLC, PA, USA</td>
			<td>BAKR8006-05</td>
			<td>diluted to 15% with normal saline for working concentration</td>
		</tr>
		<tr>
			<td>Glucose (Dianeal)</td>
			<td>Baxter International, Inc., IL, USA</td>
			<td>FNB9896</td>
			<td>Commercial dialysis solution (4.25%)</td>
		</tr>
		<tr>
			<td>GraphPad Prism 8.0</td>
			<td>GraphPad Software, Inc., CA, US</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>L-type Glu 2 assay</td>
			<td>FUJIFILM Wako, Japan</td>
			<td>461-32403</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylazine 20</td>
			<td>Juily Pharmaceutical Co., Ltd., New Taipei City, Taiwan</td>
			<td>-</td>
			<td></td>
		</tr>
		<tr>
			<td>Zoletil 50</td>
			<td>Virbac Laboratories, Carros, France</td>
			<td>-</td>
			<td></td>
		</tr>
	</tbody>
</table>

a mice model of chlorhexidine gluconate-induced peritoneal damage

Peritoneal fibrose er en vigtig komplikation af peritonealdialyse (PD). For at undersøge og løse dette problem kræves en passende dyremodel af PD. Denne protokol etablerer en chlorhexidingluconat (CG) induceret peritoneal fibrose-model, der efterligner tilstanden hos en patient med PD. Peritoneal fibrose blev induceret ved intraperitoneal injektion af 0,1% CG i 15% ethanol i 3 uger (administreret hver anden dag), i alt ni gange i mandlige C57BL/6-mus. Peritoneale funktionelle tests blev derefter udført på dag 22. Efter at musene blev ofret, blev parietal peritoneum i abdominalvæggen og leverens viscerale peritoneum høstet. De var tykkere og mere fibrotiske, når de blev analyseret mikroskopisk efter Massons trikrome farvning. Ultrafiltreringshastigheden faldt, og glukosemassetransport indikerede en CG-induceret stigning i peritonealpermeabilitet. Den således etablerede PD-model kan have anvendelser til forbedring af PD-teknologi, dialyseeffektivitet og forlængelse af patientens overlevelse.

I figur 1A, B var parietal peritoneum i abdominalvæggen markant tykkere og mere fibrotisk under Massons trikrome farvning17, hvilket indikerer, at peritoneal fibrose i den CG-eksponerede gruppe er mere alvorlig end i kontrolsaltvandsgruppen (NS). I figur 2A,B var det viscerale peritoneum på leveroverfladerne også markant tykkere og mere fibrotisk, hvilket beviser, at peritoneal fibrose i den CG-eksponerede gruppe er me...

Watch this Scientific Journal Video about A Mice Model of Chlorhexidine Gluconate-Induced Peritoneal Damage at JoVE.com

A Mice Model of Chlorhexidine Gluconate-Induced Peritoneal Damage

Denne protokol etablerer en peritonealdialyse (PD) musemodel af chlorhexidingluconat (CG)-induceret peritoneal fibrose. Den nuværende model er enkel og nem at bruge sammenlignet med andre PD dyremodeller.

En musemodel af chlorhexidingluconat-induceret peritoneal skade

Peritoneal fibrosis is an important complication of peritoneal dialysis (PD). To investigate and address this problem, an appropriate animal model of PD ...

Etablering af dyremodel for peritonealdialyse, PD, er meget vigtig. Vi håber, at det kan forbedre teknologien og effektiviteten af PD-terapi og i sidste ende forbedre patientens overlevelse. Denne peritonealdialysemusemodel er enkel, nem at bruge og synlig sammenlignet med andre peritonealdialysedyremodeller.En passende peritonealdialysedyremodel kan hjælpe forskere med at studere den peritoneale, fysiologiske og patologiske ændring under PD-terapi. Demonstration af proceduren vil være Fang-Ling Liao, en forskningsassistent fra mit laboratorium. Til at begynde med skal du forberede det kemiske irritationsmiddel ved at fortynde 0,1% chlorhexidingluconat i 15% ethanol.Tildel derefter tre mus som kontrolgruppe og injicer en milliliter pr. Kg 0,9% normalt saltvand intraperitonealt hver anden dag i tre uger i alt ni gange. Tildel derefter tre mus til peritoneal fibrosegruppen og inducer peritoneal fibrose ved at administrere en intraperitoneal injektion af 0,1% chlorhexidingluconatopløsning fremstillet tidligere i en dosis på 12,5 mikroliter pr. Kg legemsvægt. Til peritoneale funktionstest fremstilles en dialyseopløsning indeholdende 4,25%glucose.Brug derefter en sprøjte til at trække 0,5 ml dialyseprøve. Forbered en injektion af zoletil og xylazin og bedøvelse musene ved at administrere en intramuskulær injektion. Udfør intraperitoneal installation af dialyseopløsningen i en dosis på to ml pr. 20 gram legemsvægt.Lav et lille snit i midterlinjen af maven og saml den store intraperitonealvæske med en sprøjte. Efter 30 minutter skal du udføre et lodret snit i midterlinjen af maven under xiphoidprocessen og åbne musens mave for at opsamle den resterende intraperitonealvæske med en sprøjte. Mål derefter vægten af en ren og tør vatpind.Sæt bomulden i musens bughule for at absorbere den resterende intraperitonealvæske og fjern den derefter. Mål bomuldsvægten igen. Massons trikrome farvning viste, at parietal peritoneum i abdominalvæggen i den chlorhexidingluconateksponerede gruppe var markant tykkere og fibrotisk end kontrolgruppen.Det viscerale peritoneum på leveroverfladerne var også signifikant tykkere og fibrotisk i den chlorhexidingluconateksponerede gruppe end i kontrolsaltvandsgruppen, hvilket indikerer, at peritoneal fibrose er mere alvorlig i den chlorhexidingluconateksponerede gruppe. Den chlorhexidingluconateksponerede gruppe viste nedsat ultrafiltreringshastighed og øget glukosemassetransport, hvilket indikerer, at peritonealpermeabiliteten øges i chlorhexidingluconateksponerede mus Mens du udfører en intraperitoneal injektionsprocedure, skal du bruge tangen og vælge musens abdominale hud for at forhindre punkteringsinduceret intraperitoneal organskade.

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