Name: a model of chronic nutrient infusion in the rat
Uploaded: 2013-08-14T12:00:00
Description: Watch this Scientific Journal Video about A Model of Chronic Nutrient Infusion in the Rat at JoVE.com

Questo lavoro è stato sostenuto dal National Institutes of Health (R01DK58096 a Vincent Poitout). Vincent Poitout detiene il Canada Research Chair in diabete pancreatico e funzione delle cellule beta. Bader Zarrouki ricevuto borse di studio post-dottorato da Merck e Eli Lilly. Ghislaine Fontes è stato sostenuto da una borsa di studio post-dottorato della Canadian Diabetes Association.

Sommario: La procedura consiste cateterizzazione della vena giugulare destra e sinistra arteria carotide in anestesia generale; consentendo un periodo di recupero di 7 giorni; collega i cateteri alle pompe utilizzando una girevole e sistema di contrappesi che consente all&#39;animale di muoversi liberamente nella gabbia; e infondendo glucosio e / o Intralipid (un&#39;emulsione di olio di soia che genera una miscela di circa 80% unsaturated/20% di acidi grassi saturi quando infuso con eparina 9)</su...

<ol>
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C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+glucose+stimulates+early+response+gene+c-Myc+expression+in+rat+pancreatic+beta+cells.">High glucose stimulates early response gene c-Myc expression in rat pancreatic beta cells.</a> The Journal of Biological Chemistry. 276, 35375-35381 (2001).</li><li>Tang, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glucose-induced+beta+cell+dysfunction+in+vivo+in+rats:+link+between+oxidative+stress+and+endoplasmic+reticulum+stress.">Glucose-induced beta cell dysfunction in vivo in rats: link between oxidative stress and endoplasmic reticulum stress.</a> Diabetologia. 55, 1366-1379 (2012).</li><li>Alonso, L. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glucose+infusion+in+mice:+a+new+model+to+induce+beta-cell+replication.">Glucose infusion in mice: a new model to induce beta-cell replication.</a> Diabetes. 56, 1792-1801 (2007).</li><li>Magnan, C., Gilbert, M., Kahn, B. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+free+fatty+acid+infusion+in+rats+results+in+insulin+resistance+but+no+alteration+in+insulin-responsive+glucose+transporter+levels+in+skeletal+muscle.">Chronic free fatty acid infusion in rats results in insulin resistance but no alteration in insulin-responsive glucose transporter levels in skeletal muscle.</a> Lipids. 31, 1141-1149 (1996).</li><li>Goh, T. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lipid-induced+beta-cell+dysfunction+in+vivo+in+models+of+progressive+beta-cell+failure.">Lipid-induced beta-cell dysfunction in vivo in models of progressive beta-cell failure.</a> Am. J. Physiol. Endocrinol. Metab. 292, 549-560 (2007).</li><li>Pascoe, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Free+fatty+acids+block+glucose-induced+beta-cell+proliferation+in+mice+by+inducing+cell+cycle+inhibitors+p16+and+p18.">Free fatty acids block glucose-induced beta-cell proliferation in mice by inducing cell cycle inhibitors p16 and p18.</a> Diabetes. 61, 632-641 (2012).</li><li>Bell, C. G., Walley, A. J., Froguel, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+genetics+of+human+obesity.">The genetics of human obesity.</a> Nature Reviews. Genetics. 6, 221-234 (2005).</li><li>Fontes, G., Hagman, D. K., Latour, M. G., Semache, M., Poitout, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lack+of+preservation+of+insulin+gene+expression+by+a+glucagon-like+peptide+1+agonist+or+a+dipeptidyl+peptidase+4+inhibitor+in+an+in+vivo+model+of+glucolipotoxicity.">Lack of preservation of insulin gene expression by a glucagon-like peptide 1 agonist or a dipeptidyl peptidase 4 inhibitor in an in vivo model of glucolipotoxicity.</a> Diabetes Res. Clin. Pract. 87, 322-328 (2010).</li><li>Crawford, P. A., Schaffer, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metabolic+stress+in+the+myocardium:+Adaptations+of+gene+expression.">Metabolic stress in the myocardium: Adaptations of gene expression.</a> Journal of Molecular and Cellular Cardiology. , (2012).</li><li>Kewalramani, G., Bilan, P. J., Klip, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Muscle+insulin+resistance:+assault+by+lipids,+cytokines+and+local+macrophages.">Muscle insulin resistance: assault by lipids, cytokines and local macrophages.</a> Curr. Opin. Clin. Nutr. Metab Care. 13, 382-390 (2010).</li><li>Cusi, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+obesity+and+lipotoxicity+in+the+development+of+nonalcoholic+steatohepatitis:+pathophysiology+and+clinical+implications.">Role of obesity and lipotoxicity in the development of nonalcoholic steatohepatitis: pathophysiology and clinical implications.</a> Gastroenterology. 142, 711-725 (2012).</li></ol>

Sebbene un certo numero di studi precedenti hanno impiegato infusioni cronici di glucosio (ad esempio 10-15) o lipidi (ad esempio 16,17) nei roditori, a nostra conoscenza l&#39;infusione combinata di entrambi i carburanti è stato riportato soltanto nei topi 18. Il modello di infusione cronica qui presentato offre diversi vantaggi per studiare gli effetti sulla eccesso di nutrienti su una varietà di funzioni biologiche nei ratti. In primo luogo, esso non comporta rodit...

Livelli cronicamente elevati di glucosio e lipidi in circolo sono state proposte per contribuire alla patogenesi del diabete di tipo 2 alterando la funzione dei vari organi implicati nel mantenimento dell&#39;omeostasi del glucosio compreso, ma non limitato a, il pancreas cellule beta (recensione in 1). Le ipotesi postula glucotossicità che l&#39;iperglicemia cronica aggrava il difetto delle cellule beta che ha dato origine alla iperglicemia, in primo luogo, creando così un circolo vizioso e contribuendo al deterioramento del controllo glicemico nei pazienti con diabete di tipo 2. Allo stesso modo, l&#39;ipotesi glucolipotoxicity propone che aumenti concomitanti dei livelli di glucosio e di lipidi, come spesso osservati nel diabete di tipo 2, sono dannosi per la cellula beta. Decifrare i meccanismi cellulari e molecolari degli effetti deleteri di nutrienti cronicamente elevati di funzionalità delle cellule beta del pancreas è la chiave per il understanding della patogenesi del diabete di tipo 2 1. A tal fine, un gran numero di studi hanno esaminato i meccanismi della cronica nutrienti in eccesso ex vivo in isole di Langerhans isolate o in vitro in linee cellulari clonali, secernono insulina. Tuttavia, la traduzione dei risultati ottenuti in questi sistemi modello per l&#39;intero organismo è complesso, in particolare perché le concentrazioni di acidi grassi utilizzati in cellule coltivate o isolotti raramente corrispondono i livelli circolanti nelle vicinanze delle cellule beta in vivo 2. D&#39;altra parte, i meccanismi di guasto beta-cellula in risposta ad eccesso di nutrienti sono stati esaminati in modelli di roditori di diabete, come esemplificato dal ratto Zucker Diabetic Fatty 3,4, il gerbillo Psammomys obesus 5 e l&#39;alta percentuale di grassi dieta- fed del mouse 6. Questi modelli, tuttavia, sono caratterizzati da intrinseche anomalie metaboliche e non sono facilmente suscettibili di manipolazioni di glicemiae / o livelli di lipidi in un ambiente più controllato e meno cronica. Per essere in grado di alterare i livelli circolanti di nutrienti in un lasso di tempo di giorni in animali altrimenti normali, abbiamo sviluppato un modello di infusione cronica in ratti normali che ci permette di esaminare gli effetti di lipidi e del glucosio, da soli o in combinazione, su parametri fisiologici e funzionali 7,8.

<table border="1">
		<tbody>
		 <tr>
			<td>Name of Reagent/Material</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		 </tr>
		 <tr>
			<td>Saline 0.9%</td>
			<td>BD</td>
			<td>JB1324</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Dextrose 70%</td>
			<td>McKesson</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Intralipid 20%</td>
			<td>Fresenius Kabi</td>
			<td>JB6023</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Metricide (Glutaraldehyde 2.6%)</td>
			<td>Metrex</td>
			<td>11-1401</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Heparin Sodium 10,000 USP u/ml</td>
			<td>PPC</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Carprofen</td>
			<td>Metacam</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Glycopyrrolate</td>
			<td>Sandoz</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Isoflurane</td>
			<td>Abbott</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Chlohexidine 2%</td>
			<td></td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Alcohol 70%</td>
			<td></td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Iodine</td>
			<td></td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>PE-50</td>
			<td>BD</td>
			<td>427411</td>
			<td></td>
		 </tr>
		 <tr>
			<td>CO-EX T22</td>
			<td>Instech Solomon</td>
			<td>BCOEX-T22</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Connector 22G</td>
			<td>Instech Solomon</td>
			<td>SC22/15</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Swivel 22G</td>
			<td>Instech Solomon</td>
			<td>375/22PS</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Y-Connector 22G</td>
			<td>Instech Solomon</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Counterbalance and arm</td>
			<td>Instech Solomon</td>
			<td>CM375BP</td>
			<td></td>
		 </tr>
		 <tr>
			<td>23 G blunted needles</td>
			<td>Instech Solomon</td>
			<td>LS23</td>
			<td></td>
		 </tr>
		 <tr>
			<td>23 G canulation pins</td>
			<td>Instech Solomon</td>
			<td>SP23/12</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Tethers (12 inch)</td>
			<td>Lomir</td>
			<td>RT12D</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Infusion jackets </td>
			<td>Lomir</td>
			<td>RJ01, RJ02, RJ03, RJ04</td>
			<td></td>
		 </tr>
		 <tr>
			<td>(SM-XL)</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Tether attachment piece</td>
			<td>Lomir</td>
			<td>RS T1</td>
			<td></td>
		 </tr>
		 <tr>
			<td>60 ml syringe</td>
			<td>BD</td>
			<td>309653</td>
			<td></td>
		 </tr>
		 <tr>
			<td>1 ml syringe</td>
			<td>BD</td>
			<td>309602</td>
			<td></td>
		 </tr>
		</tbody>
 </table>

a model of chronic nutrient infusion in the rat

Esposizione cronica a livelli eccessivi di nutrienti è postulata a pregiudicare la funzione di diversi organi e tessuti e per contribuire allo sviluppo delle molte complicazioni associate con l&#39;obesità e la sindrome metabolica, compreso il diabete di tipo 2. Per studiare i meccanismi con cui livelli eccessivi di glucosio e acidi grassi influenzano la beta pancreatiche cellule e la secrezione di insulina, abbiamo stabilito un modello infusione di sostanze nutrienti cronica nel ratto. La procedura consiste di cateterizzazione della vena giugulare destra e sinistra arteria carotide in anestesia generale; consentendo un periodo di recupero di 7 giorni; collega i cateteri alle pompe utilizzando una girevole e sistema di contrappesi che consente all&#39;animale di muoversi liberamente nella gabbia; ed infondendo glucosio e / o Intralipid (un&#39;emulsione di olio di soia che genera una miscela di circa 80% unsaturated/20% di acidi grassi saturi quando infuso con eparina) per 72 hr. Questo modello offre diversi vantagGES, compresa la possibilità di modulare finemente i livelli target di glucosio circolante e di acidi grassi, la possibilità di co-infusione composti farmacologici, e il lasso di tempo relativamente breve rispetto a modelli alimentari. Può essere usato per esaminare i meccanismi della disfunzione nutrienti indotta in una varietà di organi e testare l&#39;efficacia dei farmaci in questo contesto.

Su una serie di 42 ratti che hanno subito un intervento chirurgico, 5 ratti sono stati persi durante il periodo post-operatorio e 1 ratto è stato perso durante l&#39;infusione, con un tasso complessivo di successo del 86%. Il peso corporeo medio dei 37 ratti che sono stati poi infuse era 608 ± 5 g prima dell&#39;intervento e 588 ± 6 g al di iniziare l&#39;infusione (media ± SE, n = 37, p &lt;0,0001 per paired t-test). I seguenti risultati rappresentativi sono stati ottenuti in due gruppi: infusione. Saline (SAL), e ...

Watch this Scientific Journal Video about A Model of Chronic Nutrient Infusion in the Rat at JoVE.com

A Model of Chronic Nutrient Infusion in the Rat

Un protocollo per infusioni cronici di glucosio e Intralipid nei ratti è descritto. Questo modello può essere utilizzato per studiare l&#39;impatto di un eccesso di nutrienti in funzione degli organi e dei parametri fisiologici.

Un modello del Chronic Infusion nutrienti nel ratto

Chronic exposure to excessive levels of nutrients is postulated to  affect the function of several organs and tissues and to contribute to the  ...

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