Name: characterization of metabolic status in nonhuman primates with the intravenous glucose tolerance test
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Forfatterne vil gerne anerkende den stærke støtte fra DHMRI CLAS dyr plejepersonalet, Facility manager Mr. Daniel Peralta og deltage dyrlæge, Dr. Glicerio Ignacio, DVM MRCVS.

Alle dyreforsøg blev godkendt af David H. Murdock Research Institute IACUC ligger på North Carolina Research Campus (NCRC), under protokol 14 til 017, Karakterisering af en ikke-human primat model af diabetes og prædiabetes / insulinresistens og effekt af lægemidler til at forbedre insulinfølsomhed og metabolisk funktion. 1. Animalske Valg og studieforberedelse <ol><li> Vælg kost og vægt stabile dyr baseret på månedlige fødeindtagelse og kropsvægt optegnelser. BEMÆRK: Dy...

<ol>
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C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Investigation+and+treatment+of+type+2+diabetes+in+nonhuman+primates.">Investigation and treatment of type 2 diabetes in nonhuman primates.</a> Methods Mol Biol. 933, 177-185 (2012).</li><li>Hansen, B. C., Bodkin, N. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Standardization+of+IVGTT.+Importance+of+method+used+to+calculate+glucose+disappearance.">Standardization of IVGTT. Importance of method used to calculate glucose disappearance.</a> Diabetes Care. 16 (5), 847 (1993).</li><li>Hardwood, J. H., Listrani, P., Wagner, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nonhuman+primates+and+other+animal+models+in+diabetes+research.">Nonhuman primates and other animal models in diabetes research.</a> J Diabetes Sci Tech. 3, 503-514 (2012).</li><li>De Koning, E. J., Bodkin, N. L., Hansen, B. C., Clark, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diabetes+mellitus+in+Macaca+mulatta+monkeys+is+characterized+by+islet+amyloidosis+and+reduction+in+beta-cell+population.">Diabetes mellitus in Macaca mulatta monkeys is characterized by islet amyloidosis and reduction in beta-cell population.</a> Diabetologia. 36, 378-384 (1993).</li><li>Letiexhe, M. R., Scheen, A. J., Gerard, P. L., Desaive, C., Lefebvre, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insulin+secretion,+clearance+and+action+before+and+after+gastroplasty+in+severely+obese+subjects.">Insulin secretion, clearance and action before and after gastroplasty in severely obese subjects.</a> Int J Obes Relat Metab Disord. 18, 295-300 (1994).</li><li>Letiexhe, M. R., Scheen, A. J., Gerard, P. L., Desaive, C., Lefebvre, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Postgastroplasty+recovery+of+ideal+body+weight+normalizes+glucose+and+insulin+metabolism+in+obese+women.">Postgastroplasty recovery of ideal body weight normalizes glucose and insulin metabolism in obese women.</a> J Clin Endocrinol Metab. 80, 364-369 (1995).</li><li>Kim, S. H., Abbasi, F., Chu, J. W., McLaughlin, T. L., Lamendola, C., Polonsky, K. S., Reaven, G. 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S., Cobelli, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+indexes+of+beta-cell+function+during+graded+up+and+down+glucose+infusion+from+C-peptide+minimal+models.">Quantitative indexes of beta-cell function during graded up and down glucose infusion from C-peptide minimal models.</a> Am J Physiol Endocrinol Metab. 280, E2-E10 (2001).</li><li>Wang, X., 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=Quantification+of+beta-cell+insulin+secretory+function+using+a+graded+glucose-infusion+with+C-peptide+deconvolution+in+dysmetabolic,+and+diabetic+cynomolgus+monkeys.">Quantification of beta-cell insulin secretory function using a graded glucose-infusion with C-peptide deconvolution in dysmetabolic, and diabetic cynomolgus monkeys.</a> Diabetology and Metabolic Syn. 5, 40 (2013).</li><li>Xiao, Y. F., Wang, B., Wang, X., Du, F., Benzinou, M., Wang, Y. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Xylazine-induced+reduction+of+tissue+sensitivity+to+insulin+leads+to+acute+hyperglycemia+in+diabetic+and+normoglycemic+monkeys.">Xylazine-induced reduction of tissue sensitivity to insulin leads to acute hyperglycemia in diabetic and normoglycemic monkeys.</a> Anesthesiology. 13 (33), (2013).</li><li>Porte, D., Kahn, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=&#946;-cell+dysfunction+and+failure+in+type+2+diabetes+potential+mechanisms.">&#946;-cell dysfunction and failure in type 2 diabetes potential mechanisms.</a> Diabetes. 50, S160-S163 (2001).</li><li>DeFronzo, R. A., Tobin, J. 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V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Serum+biochemical+changes+in+insulin+dependent+and+non-insulin+dependent+diabetes+mellitus+and+their+role+in+the+development+of+secondary+complications.">Serum biochemical changes in insulin dependent and non-insulin dependent diabetes mellitus and their role in the development of secondary complications.</a> Int J Diab Dev Countries. 17, 33-37 (1997).</li><li>Shirasaki, Y., Yoshioka, N., Kanazawa, K., Maekawa, T., Horikawa, T., Hayashi, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+physical+restraint+on+glucose+tolerance+in+cynomolgus+monkeys.">Effect of physical restraint on glucose tolerance in cynomolgus monkeys.</a> J Med Primatol. 42, 165-168 (2013).</li></ol>

Den IVGTT vurderer kapaciteten af glucose-stimuleret insulinfrigivelse af en enkelt dextrose infusion baseret på kropsvægt 5, 12, 13. Fra assayet er fastende blodglucose og insulin niveau nås, og den muliggør en vurdering af dyrets evne til at frigive insulin og returnere den forhøjede glukose niveauet mod baseline. Dette giver brugeren oplysninger at karakterisere dyr som en normal glukose og insulin niveau sund kontrol, en hyperinsulinæmisk dysmetabolic dyr med normoglykæmi, eller en hyperglykæmisk ...

Den IVGTT er en bekvem funktionelt assay, der rutinemæssigt anvendes til at bestemme β-cellefunktion hos mennesker ved forskellige metaboliske tilstande 5, 7. I dyremodeller af T2D, det er godt anerkendt som et redskab til at karakterisere dyr, der viser metabolisk sygdomsprogression fra en sundt at en dysmetabolic hyperglykæmisk tilstand 8, 9. Den nærmeste dyremodel af T2D demonstreres i ikke-humane primater (menneskelige primater), hvoraf rhesus og cynomolgus makakaber er bemærkelsesværdige eksempler. Disse dyr naturligt udvikle T2D med de samme risikofaktorer alder og fedme bidrager til forekomsten som hos mennesker 10. Endvidere er der en lignende sygdomsprogression og pancreas patologi viser amyloidaflejringer som dysmetabolic sygdommen skrider 11. Her rapporterer vi på vores standard metode til at udføre en IVGTT i NHP&#39;er som en del af vores koloni karakterisering af metaboliske status i disse dyr. Denne metode erlet at udføre i forhold til andre, mere tidskrævende og kostbare teknikker 2. Den IVGTT er nyttig til karakterisering en stor koloni af dyrene hurtigt og ofte. Når det tages i betragtning med niveauet for glyceret hæmoglobin (HbA1c), dyrets kost og fødeindtagelse historie, samt deres procent mager masse og kropsfedt, den IVGTT er normalt tilstrækkeligt til at karakterisere et dyrs metaboliske status og progression mod åbenlys diabetes 6 , 8. HbA1C repræsenterer den gennemsnitlige glykæmiske niveau over livet i en røde blodlegemer, hvilket giver en pålidelig måling af blodsukkerniveauet i de foregående seks uger til tre måneder. Målt fra fastende baseline blodprøve af IVGTT, denne værdi giver et vindue til glykæmisk kontrol i løbet af måneder mellem procedurer. Hvis dyret har skiftet fra dysmetabolic til diabetisk siden deres sidste IVGTT, ville en HbA1C-værdi langt højere end deres tidligere værdi angiverat overgangen begyndte kort efter deres sidste IVGTT, mens en HbA1C værdi tættere på deres tidligere værdi tyder på, at de først for nylig har skiftet. Generelt i makakaber, HbA1C-værdier større end 6%, betragtes unormal, og indikerer dårlig glykæmisk kontrol 10, 23. Glykæmiske niveauer bør fortolkes i sammenhæng med den adfærd og generelle sundhed af dyret som helhed. Diabetiske makakaber - ligesom mennesker - udstille hyperfagi, polydipsi og polyuri. Gruppe opstaldning af dyr giver væsentlige udfordringer for måling af disse indikatorer og det individuel pleje kræves til dysmetabolic og diabetiske aber. Vi anbefaler enkeltvis boliger dyrene for at mere personlig pleje kan leveres, og adfærdsmæssige markører for sundhed aben lettere overvåges 8. Derudover vil diabetiske makakaber udviser vægttab, samt en forhøjet lipidprofil (øgetkolesterol, hypertriglyceridæmi) og forstyrret mineralmetabolismen i serum kemi. Det er vigtigt at måle markører for lever- og nyrefunktion i serum kemi, som skade på disse organer er ofte komplikationer af fremrykkende stofskiftesygdom / diabetes, og kan co-determinanter for glykæmisk, lipid og mineralske ubalancer 9, 11, 18, 24 . Når du bruger denne metode, de historiske værdier genereret fra flere, hyppige karakteriseringer løbet af livet af en abe er af særlig værdi. Hvis andre procedurer, såsom en glukose clamp eller gradueret glukose infusion (GGI), er nødvendige for at vurdere et dyrs sundhed, er det almindeligt ved første karakterisering når deres historie er ikke tilgængelig. Men når en baseline er blevet etableret, gentagne IVGTTs af en frekvens på hver tredje måned normalt tilstrækkelige til at spore et dyrs udvikling. Dette er især vigtigt, når dyrene er tilmeldt i flere studier i hele enkalenderår baseret på deres metaboliske status. Mens deres helbred kan forblive relativt stabil i år ad gangen, når den metaboliske status af et dyr forværres, kan en dramatisk stigning i insulinresistens og glucoseintolerans forekomme meget hurtigt. HbA1C værdier tillader nogle interpolation af faldet eller forbedring af sundhedstilstanden af ​​dyret mellem procedurer planlagt tre måneders mellemrum. Af denne grund er denne metode ideel til karakterisering dyr anvendt i multiple, langsgående undersøgelser i løbet af deres naturlige levetid.

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			<td height="42" style="height:42px;width:216px;">Allegra X-15R Centrifuge</td>
			<td style="width:147px;"></td>
			<td style="width:119px;"></td>
			<td style="width:188px;">plasma: 4C @3000 rpm for 10 min</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">Sorvall ST16R Centrifuge</td>
			<td></td>
			<td style="width:119px;"></td>
			<td style="width:188px;">serum: 22C @3000 rpm for 10 min</td>
		</tr>
		<tr height="65">
			<td height="65" style="height:65px;width:216px;">Thermo Scientific -86C Freezer, Forma 88000 Series</td>
			<td></td>
			<td>Model: 88500A</td>
			<td style="width:188px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Dextrose 50% (D50)</td>
			<td style="width:147px;">Webster</td>
			<td style="width:119px;">07-8008986</td>
			<td style="width:188px;">I.V. glucose infusate</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">3mL Luer Lock Syringe</td>
			<td style="width:147px;">Midwest Veterinary Suppy</td>
			<td style="width:119px;"></td>
			<td style="width:188px;">serial blood draws</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">5ml Luer Lock Syringe</td>
			<td style="width:147px;">Midwest Veterinary Suppy</td>
			<td style="width:119px;"></td>
			<td style="width:188px;">heparinized saline flush</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">10mL Luer Lock Syringe</td>
			<td style="width:147px;">Midwest Veterinary Suppy</td>
			<td style="width:119px;"></td>
			<td style="width:188px;">delivery of I.V. D50</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Gauze sponges 2x2</td>
			<td style="width:147px;">Midwest Veterinary Suppy</td>
			<td style="width:119px;">366.23000.4</td>
			<td style="width:188px;">Used Dry, w/ %70 Alcohol, and 2% Chlorohex Solution</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">4 ml serum separator tubes&#160;</td>
			<td style="width:147px;">Midwest Veterinary Supply</td>
			<td style="width:119px;">366.45000.4</td>
			<td style="width:188px;">blood collection tube for superchem panel</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">K2EDTA, 2mL</td>
			<td style="width:147px;">VWR</td>
			<td style="width:119px;">95057-239</td>
			<td style="width:188px;">blood collection tubes</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Aprotinin, 100mg</td>
			<td style="width:147px;">Sigma</td>
			<td style="width:119px;">A1153-100MG</td>
			<td style="width:188px;">blood collection tube protease additive</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">22g x 1&#34; Catheters</td>
			<td style="width:147px;">Midwest Veterinary Suppy</td>
			<td style="width:119px;">193.75250.2</td>
			<td style="width:188px;">I.V. catheter&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Injection Plug W/ Cap</td>
			<td style="width:147px;">Midwest Veterinary Suppy</td>
			<td style="width:119px;">001.11500.2</td>
			<td style="width:188px;">%50 dextrose infusion port</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Porus Tape, 1/2&#34; x 10yd&#160;</td>
			<td style="width:147px;">Midwest Veterinary Suppy</td>
			<td style="width:119px;">001.85000.2</td>
			<td style="width:188px;">maintain adherance of catheters and hep. Locks</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Chlorhexidine Solution 2%</td>
			<td style="width:147px;">Midwest Veterinary Suppy</td>
			<td style="width:119px;">193.08855.3</td>
			<td style="width:188px;">prep catheter site</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">70% Ethanol</td>
			<td style="width:147px;">VWR</td>
			<td style="width:119px;">71001-654</td>
			<td style="width:188px;">prep catheter site</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">tourniquet</td>
			<td style="width:147px;">Webster</td>
			<td style="width:119px;">07-8003432</td>
			<td style="width:188px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">3 way stopcock</td>
			<td style="width:147px;">Midwest Veterinary Supply</td>
			<td style="width:119px;">366.28510.4</td>
			<td style="width:188px;">hep. lock</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">37&#34; extension set</td>
			<td style="width:147px;">Webster</td>
			<td style="width:119px;">07-8454200</td>
			<td style="width:188px;">hep. lock</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Exel 50-60cc LL Syringes</td>
			<td style="width:147px;">Midwest Veterinary Suppy</td>
			<td style="width:119px;">001.12250.2</td>
			<td style="width:188px;">Heparinized saline flush</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">250 ml bag 0.9% saline</td>
			<td style="width:147px;">Webster</td>
			<td style="width:119px;">07-8365593</td>
			<td style="width:188px;">flush</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">1,000 U Heparin, 10 ml</td>
			<td style="width:147px;">Webster</td>
			<td style="width:119px;">07-883-4916</td>
			<td style="width:188px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Ketamine, (Ketaset) 100mg/mL</td>
			<td style="width:147px;">Fort Dodge</td>
			<td style="width:119px;">(AV ordered)</td>
			<td style="width:188px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Precision Xtra glucose test strips 50/bx</td>
			<td style="width:147px;">Abbott (American Diabetes Wholesale)</td>
			<td style="width:119px;">9381599728K7</td>
			<td style="width:188px;">test baseline/ T3 blood glucose levels</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Masimo Rad 57</td>
			<td style="width:147px;">DRE</td>
			<td style="width:119px;">6052057V</td>
			<td style="width:188px;">pulse-oximeter</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Pavia rectal thermometer</td>
			<td style="width:147px;">Patterson</td>
			<td style="width:119px;">07-8391335</td>
			<td style="width:188px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Precision Xtra Glucometer</td>
			<td style="width:147px;">Abbott</td>
			<td style="width:119px;">9381599728K7</td>
			<td style="width:188px;">Handheld glucometer</td>
		</tr>
	</tbody>
</table>

characterization of metabolic status in nonhuman primates with the intravenous glucose tolerance test

Den intravenøse glucose tolerance test (IVGTT) spiller en central rolle i karakterisering af glukose homøostase. Når det tages sammen med serum biokemiske profiler, inklusive blodsukkerniveauet i både fodres og fastende tilstand, HbA1c, insulin niveauer, klinisk historie kost, kropssammensætning og kropsvægt status, en vurdering af normal og unormal glykæmisk kontrol kan foretages . Fortolkning af en IVGTT sker gennem målingen af ​​ændringer i glucose- og insulinniveauer over tid i forhold til dextrose udfordring. Kritiske komponenter, der skal overvejes, er: peak glucose og insulin niveauer nået i forhold til T0 (slutningen af glukose infusion), glucose clearance K afledt fra hældningen af hurtige glukose clearance i de første 20 min (T1 til T20), den tid, at vende tilbage til glucose baseline, og arealet under kurven (AUC). Disse IVGTT foranstaltninger viser karakteristiske ændringer som glucosehomeostase bevæge sig fra en sund toa syg metaboliske tilstand 5. Heri beskriver vi karakteriseringen af ​​humane primater (rhesus og Cynomolgus makakaber), som er mest relevante dyremodel for type II-diabetes (T2D) hos mennesker og IVGTT og kliniske profiler af disse dyr fra et magert sundt, at fede dysmetabolic, og T2D tilstand 8, 10, 11.

De i figur 1 resultaterne er demonstrative af typiske glucose og insulin Kurverne fra modne, sunde og diabetiske cynomolgus makakaber i løbet af en 30 min IVGTT. Data fra raske og avancerede diabetiske aber er vist for at kontrastere de indlysende forskelle mellem dyr fra begge yderste ender af rækken af ​​metaboliske karakterisering. Denne IVGTT protokol er blevet anvendt med succes af forfatterne i rhesus makakaber med lignende resultater. <p class="jove_cont...

Watch this Scientific Journal Video about Characterization of Metabolic Status in Nonhuman Primates with the Intravenous Glucose Tolerance Test at JoVE.com

Characterization of Metabolic Status in Nonhuman Primates with the Intravenous Glucose Tolerance Test

Målet med denne protokol er at præsentere en standard metode til at udføre intravenøs glukose tolerance test (IVGTTs) for at vurdere glykæmisk kontrol i ikke-humane primater og vurdere deres metaboliske status fra sundt at dysmetabolic.

Karakterisering af metaboliske status i ikke-humane primater med intravenøs glukose Tolerance Test

The intravenous glucose tolerance test (IVGTT) plays a key role in the characterization of glucose homeostasis. When taken together with serum biochemical ...

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