Name: intraperitoneal glucose tolerance test, measurement of lung function, and fixation of the lung to study the impact of obesity and impaired metabolism on pulmonary outcomes
Uploaded: 2018-03-15T13:00:00
Description: Watch this Scientific Journal Video about Intraperitoneal Glucose Tolerance Test, Measurement of Lung Function, and Fixation of the Lung to Study the Impact of Obesity and Impaired Metabolism on Pulmo

Forsøgene blev støttet af Marga og Walter Boll-Stiftung, Kerpen, Tyskland; Projekt 210-02-16 (MAAA), projekt 210-03-15 (MAAA) og af den tyske Forskningsfonds (DFG; AL1632-02; MAAA), Bonn, Tyskland. Center for Molekylær medicin Köln (CMMC; Universitetshospital Köln; Karriere avancement Program; MAAA), Köln formue (fakultetet for medicin, universitetet i Köln; KD).

Alle dyr, der blev gennemført i overensstemmelse med protokoller godkendt af lokale myndigheder (delstaten NRW, AZ: 2012.A424), og var i overensstemmelse med den tyske dyrevelfærd lov og forordninger om velfærd for dyr, der anvendes for eksperimenter eller for andre videnskabelige formål. Da lunge-funktionen analyse kan påvirke lunge struktur og derfor efterfølgende histologisk analyserer, bør måling af Res og Cdyn og forberedelse og fiksering af lunge for histomorfometri udføres i forskellige dyr. Måling af Re...

<ol>
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Denne rapport indeholder tre protokoller for tre forskellige metoder til at analysere virkningen af fedme på glukosemetabolismen og pulmonal resultater. Først, den glukose tolerance test giver mulighed for at analysere intracellulære glukoseoptagelse og kan være vejledende for insulinresistens. Andet, hele kroppen plethysmography er en teknik til at måle lungefunktion og dermed nyttigt at teste effekten af nye behandlinger. For det tredje er en standardiseret fiksering protokol afgørende for kvantitative morfometri...

Ifølge World Health Organization (WHO) i 2008, var mere end 1,4 milliarder voksne, i alderen 20 og ældre, overvægtig med et body mass index (BMI) større end eller lig med 25; yderligere, mere end 200 millioner mænd og næsten 300 millioner kvinder var overvægtige (BMI≥30)1. Fedme og metaboliske syndrom er vigtige risikofaktorer for en lang række sygdomme. Mens fedme og samtidig øget hvidt fedtvæv masse har været tæt forbundet med type 2 diabetes2,3, hjerte-kar sygdomme, herunder koronar hjertesygdom (CHD), hjertesvigt (HF), atrieflimren4 og slidgigt5, deres funktionelle roller i patogenesen af luftvejslidelser forbliver dårligt forstået. Dog har epidemiologiske undersøgelser påvist, at fedme er stærkt forbundet med kroniske luftvejslidelser, herunder anstrengelses dyspnø, obstruktiv søvnapnø syndrom (OSAS), fedme hypoventilation syndrom (OHS), kronisk obstruktiv lungesygdom (KOL), lungeemboli, aspirationspneumoni og bronkial astma6,7,8,9. Potentielle mekanismer forbinder fedme og foruroliget stofskifte, fx, insulinresistens og type II diabetes, til patogenesen af kronisk lungesygdom ikke kun omfatter mekaniske og fysiske konsekvenser af vægt gevinst ved ventilation men også fremkalde en kronisk subakutte inflammatorisk tilstand10,11. Stigningen i fedme og lungesygdomme i det sidste tiår, kombineret med manglen på effektive forebyggende strategier og terapeutiske tilgange, fremhæver nødvendigheden af at undersøge de molekylære mekanismer for at definere nye muligheder for at styre fedme-relaterede lungekræft sygdomme.
Her, vi beskriver tre standard tests, som er vigtige grundlæggende at undersøge fedme og dens indvirkning på lunge struktur og funktion i musemodeller: (1) intraperitoneal glukose tolerance (ipGTT) (2) måling af luftvejene modstand (Res) og respiratorisk systemets kompatibilitet (Cdyn); og (3) forberedelse og fiksering af lunge for efterfølgende kvantitativ histologisk vurdering. IpGTT er en robust screeningstest foranstaltning glukoseoptagelse, og dermed effekten af fedme på stofskiftet. Enkelheden i metoden tillader god standardisering, og derfor sammenlignelighed af resultaterne mellem laboratorier. Mere sofistikerede metoder, såsom hyperglykæmisk klemmer eller undersøgelser på isolerede småøer, kan bruges til detaljeret analyse af metabolisk fænotype12. Her vurderer vi glukosetolerance for at definere en fedme-associeret stat af systemisk og metabolisk lidelse som grundlag for yderligere undersøgelser af en pulmonal resultatet. For at vurdere effekten af fedme og stofskiftesygdom på lungefunktion, målte vi luftvejene modstand (Res) og åndedrætsorganerne overholdelse (Cdyn). For at karakterisere lungesygdom, er uhæmmet samt behersket metoder til vurdering af lungefunktion tilgængelige. Uhæmmede plethysmography i frit flytte dyr efterligner en naturlig tilstand, som afspejler vejrtrækning mønstre; invasive metoder, såsom Indgangsimpedans måling af Res og cDyn i dybt bedøvede mus at vurdere dynamisk lunge mekanik, er derimod mere præcis13. Da kroniske luftvejssygdomme afspejles af histologiske ændringer af lungevæv, er ordentlig lunge fiksering til yderligere analyse forestående. Valget af metode til væv fiksering og forberedelse afhænger rum i lungerne, som vil blive undersøgt, for eksempel, gennemføre airways eller lunge parenkym14. Her, beskriver vi en metode, der giver mulighed for kvalitativ og kvantitativ vurdering af de strømførende airways at studere effekten af fedme på astma udvikling.

<table>
	<tbody>
		<tr>
			<td>GlucoMen LX</td>
			<td>A.Menarini diagnostics, Firneze, Italy</td>
			<td>38969</td>
			<td>blood glucose meter</td>
		</tr>
		<tr>
			<td>GlucoMen LX Sensor</td>
			<td>A.Menarini diagnostics, Firneze, Italy</td>
			<td>39765</td>
			<td>Test stripes</td>
		</tr>
		<tr>
			<td>Glucose 20%</td>
			<td>B. Braun, Melsung, Germany</td>
			<td>2356746</td>
			<td></td>
		</tr>
		<tr>
			<td>FinePointe Software</td>
			<td>DSI, MC s&#180;Hertogenbosch, Netherlands</td>
			<td>601-1831-002</td>
			<td></td>
		</tr>
		<tr>
			<td>FinePointe RC Single Site Mouse Table</td>
			<td>DSI, MC s&#180;Hertogenbosch, Netherlands</td>
			<td>601-1831-001</td>
			<td></td>
		</tr>
		<tr>
			<td>FPRC Controller</td>
			<td>DSI, MC s&#180;Hertogenbosch, Netherlands</td>
			<td>601-1075-001</td>
			<td></td>
		</tr>
		<tr>
			<td>FPRC Aerosol Block</td>
			<td>DSI, MC s&#180;Hertogenbosch, Netherlands</td>
			<td>601-1106-001</td>
			<td></td>
		</tr>
		<tr>
			<td>Aerogen neb head-5.2-4um</td>
			<td>DSI, MC s&#180;Hertogenbosch, Netherlands</td>
			<td>601-2306-001</td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps</td>
			<td>FST, British Columbia, Canada</td>
			<td>11065-07</td>
			<td></td>
		</tr>
		<tr>
			<td>Blunt scissors</td>
			<td>FST, British Columbia, Canada</td>
			<td>14105-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro scissors</td>
			<td>FST, British Columbia, Canada</td>
			<td>15000-00</td>
			<td></td>
		</tr>
		<tr>
			<td>Perma-Hand 4-0</td>
			<td>Ethicon, Puerto Rico, USA</td>
			<td>736H</td>
			<td>Surgical suture</td>
		</tr>
		<tr>
			<td>Roti-Histofix 4%</td>
			<td>Roth</td>
			<td>P087.1</td>
			<td>4% Paraformaldehyd</td>
		</tr>
		<tr>
			<td>Ketaset</td>
			<td>Zoetis, Berlin, Germany</td>
			<td>10013389</td>
			<td>Ketamine</td>
		</tr>
		<tr>
			<td>Rompun 2%</td>
			<td>Bayer, Leverkusen, Germany</td>
			<td>770081</td>
			<td>Xylazine</td>
		</tr>
	</tbody>
</table>

intraperitoneal glucose tolerance test, measurement of lung function, and fixation of the lung to study the impact of obesity and impaired metabolism on pulmonary outcomes

Fedme og luftvejssygdomme er store sundhedsmæssige problemer. Fedme er blevet en ny epidemi med et forventet antal over 1 milliard overvægtige personer på verdensplan frem til 2030, således udgør en voksende samfundsøkonomisk byrde. Samtidig, er fedme-relaterede co-morbiditet, herunder diabetes og hjerte samt kroniske lungesygdomme, konstant stigende. Selv om fedme er forbundet med øget risiko for astma-eksacerbationer, forværring af respiratorisk symptomer og dårlig kontrol, funktionelle rolle fedme og foruroliget metabolisme i patogenesen af kronisk lungesygdom er ofte undervurderet, og underliggende molekylære mekanismer er fortsat undvigende. Denne artikel har til formål at præsentere metoder til at vurdere effekten af fedme på stofskifte, samt lunge struktur og funktion. Her, vi beskriver tre teknikker for mus undersøgelser: (1) vurdering af intraperitoneal glukosetolerance (ipGTT) at analysere effekten af fedme på glukosemetabolismen; (2) måling af luftvejene modstand (Res) og åndedrætsorganerne overholdelse (Cdyn) at analysere effekten af fedme på lungefunktion; og (3) forberedelse og fiksering af lunge for efterfølgende kvantitativ histologisk vurdering. Fedme-relaterede lungesygdom er sandsynligvis multifaktoriel, skyldes systemisk inflammatorisk og metaboliske dysregulering, der potentielt negativt påvirker lungefunktion og respons på terapi. En standardiseret metode til at studere molekylære mekanismer og effekt af nye behandlinger er derfor afgørende.

Repræsentative resultater af intraperitoneal glukose tolerance test (ipGTT) (figur 4), lunge-funktion test (figur 5), og repræsentant billeder illustrerer hæmatoxylin og eosin farves lungerne (figur 6).
IpGTT blev udført på overvægtige mus (blå) efter 7 uger af høj-fedt-kost (HFD). Standard kost-fed mus fungerede som kontrolelemente...

Watch this Scientific Journal Video about Intraperitoneal Glucose Tolerance Test, Measurement of Lung Function, and Fixation of the Lung to Study the Impact of Obesity and Impaired Metabolism on Pulmo

Intraperitoneal Glucose Tolerance Test, Measurement of Lung Function, and Fixation of the Lung to Study the Impact of Obesity and Impaired Metabolism on Pulmonary Outcomes

Forekomsten af fedme er stigende og øger risikoen for kroniske lungesygdomme. For at fastsætte de underliggende mekanismer og forebyggende strategier, veldefinerede dyre modeller er nødvendige. Her give vi tre metoder (glucose-tolerance-test, krop plethysmography og lunge fiksering) at studere effekten af fedme på pulmonal resultater i mus.

Intraperitoneal glukose Tolerance Test, måling af lungefunktion og fiksering af lungerne at studere virkningerne af fedme og hæmmet metabolisme på pulmonal resultater

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