Name: laser doppler: a tool for measuring pancreatic islet microvascular vasomotion in vivo
Uploaded: 2018-03-08T13:00:00
Description: Watch this Scientific Journal Video about Laser Doppler: A Tool for Measuring Pancreatic Islet Microvascular Vasomotion In Vivo at JoVE.com

Dette arbejde blev støttet af tilskud fra Peking Union Medical College Ungdom fonden og den grundlæggende forskningsmidler til Central universiteter (Grant nr. 3332015200).

Alle dyreforsøg blev henrettet i overensstemmelse med alle relevante retningslinjer, regler og reguleringsorganer. Denne protokol bliver demonstreret blev udført under vejledning og godkendelse af de Institute af mikrocirkulationen dyr etiske udvalg (IMAEC) på Peking Union Medical College (PUMC).
1. dyr
<ol><li>Før starten af eksperimentet, holde tre BALB/c mus pr. bur med kontrolleret temperatur (24 ± 1 ° C) og fugtighed (55 ± 5%), under en 12-h lys-mørke cyklus. Give musene gratis ...

<ol>
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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=Retinal+blood+flow+is+increased+in+type+1+diabetes+mellitus+patients+with+advanced+stages+of+retinopathy.">Retinal blood flow is increased in type 1 diabetes mellitus patients with advanced stages of retinopathy.</a> BMC Endocr Disord. 16 (1), 25-33 (2016).</li><li>Forst, 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=Retinal+Microcirculation+in+Type+1+Diabetic+Patients+With+and+Without+Peripheral+Sensory+Neuropathy.">Retinal Microcirculation in Type 1 Diabetic Patients With and Without Peripheral Sensory Neuropathy.</a> J Diabetes Sci Technol. 8 (2), 356-361 (2014).</li><li>Hu, H. F., Hsiu, H., Sung, C. J., Lee, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Combining+laser-Doppler+flowmetry+measurements+with+spectral+analysis+to+study+different+microcirculatory+effects+in+human+prediabetic+and+diabetic+subjects.">Combining laser-Doppler flowmetry measurements with spectral analysis to study different microcirculatory effects in human prediabetic and diabetic subjects.</a> Lasers Med Sci. 31 (1), 1-8 (2016).</li><li>Klonizakis, M., Manning, G., Lingam, K., Donnelly, R., Yeung, J. 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I de tilfælde, der involverer mikrovaskulære dysfunktion (f.eks. diabetes, akut pancreatitis, perifere mikrovaskulære sygdomme, osv.), føre visse sygdomme til nedsat blodgennemstrømning. Bortset fra ændringer i blodgennemstrømningen er der vigtige indikatorer, såsom mikrovaskulære hjerte, der afspejler den funktionelle status af mikrocirkulationen. Den specifikke indikator, mikrovaskulære hjerte, defineres normalt som svingningerne i mikrovaskulære tonen i mikrovaskulære senge. I den nuvære...

Som en parameter i den funktionelle status af mikrocirkulationen, mikrovaskulære hjerte tager ansvar for levering og udveksling af ilt, næringsstoffer og hormoner og er afgørende for fjernelse af metaboliske produkter, såsom kuldioxid og celle affald 1. mikrovaskulære hjerte også regulerer blod flow distribution og væv perfusion, derved påvirker lokale microcirculatory blodtryk og svar til inflammation, som kan fremkalde ødem i mange sygdomme. Mikrovaskulære hjerte er derfor yderst vigtigt at opretholde den fysiologiske funktion af organer2,3,4, væv og celler, komponent. Værdiforringelse af mikrovaskulære hjerte kan være en af de vigtigste skridt i udviklingen af mikrocirkulationen-relaterede sygdomme5.
Laser Doppler blev oprindeligt udviklet til observation og kvantificering i feltet af mikrocirkulationen forskning6. Denne teknik, sammen med andre tekniske tilgange (f.eks. laser speckle7, transkutan ilt, osv.) er blevet betragtet som den gyldne standard for vurdering af blodgennemstrømningen i mikrocirkulationen. Rationalet at blod perfusion af lokale mikrocirkulationen (dvs. kapillærer, arterioler, venules, etc.) kan bestemmes ved apparatet udstyret med laser Doppler, er baseret på Doppler Skift princip. Bølgelængde og frekvens af stimuleret emission lys ændre når lys partikler støder bevægelige blodlegemer i microvessels, eller de forbliver uændrede. Derfor, i mikrocirkulationen, antallet og hastigheden af blodlegemer er de vigtigste faktorer vedrørende omfang og hyppighed distribution af Doppler-skiftet lyset, mens retningen af mikrovaskulære blodgennemstrømningen er irrelevant. Ved hjælp af forskellige metoder, en bred vifte af væv har været brugt i microcirculatory undersøgelser, herunder mesenteries og dorsal skinfold kamre af mus, rotter, hamstere, og endda mennesker8. Men i den nuværende protokol, vi fokuserer på den funktionelle status af pancreas islet mikrovaskulære hjerte, som evalueres ved hjælp af laser Doppler og en hjemmelavet parameter evalueringssystem.
Pancreas islet mikrocirkulationen er hovedsageligt sammensat af pancreas Holmen microvessels og udstiller særpræg. En bugspytkirtlen islet kapillære netværk viser en fem gange højere tæthed end den kapillære netværk af dens eksokrine modstykke9. At give en kanal for levering af input glukose og udbrede insulin, levere Holmen endotelceller ilt til metabolisk aktive celler i Holmen β celler. Desuden viser nye beviser også at Holmen microvessels er involveret i reguleringen af insulin Gen-ekspression og β-celle overlevelse, men også i påvirker β-celle funktion; fremme af β-celle spredning; og producerer en række vasoaktive, angiogene stoffer og vækstfaktorer10. Derfor i denne henseende udlede vi at den funktionelle status af pancreas islet mikrovaskulære hjerte kan påvirke islet β-celle funktion og blive involveret i patogenesen af sygdomme som akut/kronisk betændelse i bugspytkirtlen, diabetes og andre bugspytkirtel-relaterede sygdomme.
Analysere de patologiske ændringer af den funktionelle status af pancreas islet mikrovaskulære hjerte kan være en mulig strategi at bestemme bidrag af pancreas islet mikrocirkulationen til de sygdomme, der er nævnt ovenfor. En detaljeret, trinvis fremgangsmåde beskriver den tilgang til at bestemme i bugspytkirtlen islet mikrovaskulære hjerte i vivo give her. Typiske målinger vises derefter i Repræsentative resultater. Endelig, fordele og begrænsninger af metoden, der er fremhævet i diskussionen, sammen med yderligere programmer.

<table>
	<tbody>
		<tr>
			<td>MoorVMS-LDF2</td>
			<td>Moor Instruments</td>
			<td>GI80</td>
			<td>PeriFlux 5000 (Perimed Inc.) can be used as an alternative apparatus to harvest data</td>
		</tr>
		<tr>
			<td>MoorVMS-PC Software</td>
			<td>Moor Instruments</td>
			<td>GI80-1</td>
			<td>Software of MoorVMS-LDF2</td>
		</tr>
		<tr>
			<td>Calibration stand</td>
			<td>Moor Instruments</td>
			<td>GI-cal</td>
			<td>Calibration tool</td>
		</tr>
		<tr>
			<td>Calibration base</td>
			<td>Moor Instruments</td>
			<td>GI-cal</td>
			<td>Calibration tool</td>
		</tr>
		<tr>
			<td>Calibration flux standard</td>
			<td>Moor Instruments</td>
			<td>GI-cal</td>
			<td>Calibration tool</td>
		</tr>
		<tr>
			<td>One Touch UltraEasy glucometer</td>
			<td>Johnson and Johnson</td>
			<td>#1955685</td>
			<td>Confirm hyperglycemia</td>
		</tr>
		<tr>
			<td>One Touch UltraEasy strips</td>
			<td>Johnson and Johnson</td>
			<td>#1297006</td>
			<td>Confirm hyperglycemia</td>
		</tr>
		<tr>
			<td>Streptozotocin</td>
			<td>Sigma-Aldrich</td>
			<td>S0130</td>
			<td>Dissolve in sodium citrate buffer (pH 4.3)</td>
		</tr>
		<tr>
			<td>Pentobarbital sodium</td>
			<td>Sigma-Aldrich</td>
			<td>P3761</td>
			<td>Working concentration 3 %</td>
		</tr>
		<tr>
			<td>Ethanol</td>
			<td>Sinopharm Inc.</td>
			<td>200121</td>
			<td>Working concentration 75 %</td>
		</tr>
		<tr>
			<td>Sucrose</td>
			<td>Amresco</td>
			<td>335</td>
			<td>Working concentration 10 %</td>
		</tr>
		<tr>
			<td>Medical gauze</td>
			<td>China Health Materials Co.</td>
			<td>S-7112</td>
			<td>Surgical</td>
		</tr>
		<tr>
			<td>Blunt-nose forceps</td>
			<td>Shang Hai Surgical Instruments Inc.</td>
			<td>N-551</td>
			<td>Surgical</td>
		</tr>
		<tr>
			<td>Surgical tapes</td>
			<td>3M Company</td>
			<td>3664CU</td>
			<td>Surgical</td>
		</tr>
		<tr>
			<td>Gauze sponge</td>
			<td>Fu Kang Sen Medical Device CO.</td>
			<td>BB5447</td>
			<td>Surgical</td>
		</tr>
		<tr>
			<td>Scalpel</td>
			<td>Yu Lin Surgical Instruments Inc.</td>
			<td>175C</td>
			<td>Surgical</td>
		</tr>
		<tr>
			<td>Skin scissor</td>
			<td>Carent</td>
			<td>255-17</td>
			<td>Surgical</td>
		</tr>
		<tr>
			<td>Suture</td>
			<td>Ning Bo Surgical Instruments Inc.</td>
			<td>3325-77</td>
			<td>Surgical</td>
		</tr>
		<tr>
			<td>Syringe and 25-G needle</td>
			<td>MISAWA Inc.</td>
			<td>3731-2011</td>
			<td>Scale: 1 ml</td>
		</tr>
	</tbody>
</table>

laser doppler: a tool for measuring pancreatic islet microvascular vasomotion in vivo

Som funktionelle status af mikrocirkulationen er mikrovaskulære hjerte vigtigt for levering af ilt og næringsstoffer, og fjernelse af kuldioxid og affaldsprodukter. Værdiforringelse af mikrovaskulære hjerte kan være et afgørende skridt i udviklingen af mikrocirkulationen-relaterede sygdomme. Derudover er meget vaskulariserede pancreas Holmen tilpasset til at støtte endokrine funktion. I denne henseende forekommer det muligt at udlede, at den funktionelle status af pancreas islet mikrovaskulære hjerte kan påvirke pancreas islet funktion. Analysere de patologiske ændringer af den funktionelle status af pancreas islet mikrovaskulære hjerte kan være en mulig strategi at bestemme bidrag at pancreas islet mikrocirkulationen gør til relaterede sygdomme som diabetes mellitus, pancreatitis, osv. Derfor, denne protokol beskriver ved hjælp af en laser Doppler blod flow skærm til at bestemme den funktionelle status af pancreas islet mikrovaskulære hjerte, og at fastsætte parametre (herunder gennemsnitlige blod perfusion, amplitude, frekvens og relativ hastigheden af pancreas islet mikrovaskulære hjerte) til evaluering af microcirculatory funktionelle status. I en streptozotocin-induceret diabetisk musemodel konstaterede vi en nedsat funktionel status af pancreas islet mikrovaskulære hjerte. Afslutningsvis kan denne tilgang til vurdering af pancreas islet mikrovaskulære hjerte i vivo afsløre mekanismer vedrørende pancreas islet sygdomme.

Et fotografi af mikrovaskulære hjerte måling laser Doppler apparatet udstyret med en halvleder laserdiode er vist i figur 1A. Bruger interface software er præsenteret i figur 1B. Ved hjælp af den metode nævnt ovenfor, blev de hæmodynamiske parametre af pancreas islet mikrovaskulære hjerte opdaget for både ikke-diabetisk kontrol og diabetisk mus. En række teknikker, herunder laser Doppler flowmetry, afspejles og spredt ly...

Watch this Scientific Journal Video about Laser Doppler: A Tool for Measuring Pancreatic Islet Microvascular Vasomotion In Vivo at JoVE.com

Laser Doppler: A Tool for Measuring Pancreatic Islet Microvascular Vasomotion In Vivo

Laser Doppler: Et værktøj til måling af pancreas Islet mikrovaskulære hjerte In Vivo

Pancreas islet mikrovaskulære hjerte regulerer islet blod distribution og vedligeholder den fysiologiske funktion af holmen β celler. Denne protokol beskriver ved hjælp af en laser Doppler skærm til at bestemme den funktionelle status af pancreas islet mikrovaskulære hjerte i vivo og at vurdere bidragene fra pancreas islet mikrocirkulationen pancreas-relaterede sygdomme.

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Laser Doppler: A Tool for Measuring Pancreatic Islet Microvascular Vasomotion In Vivo

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The microcirculation consists of blood vessels with diameters less than 150 &#181;m. It makes up a large part of the circulatory system and plays an ...

Measuring Ascending Aortic Stiffness In Vivo in Mice Using Ultrasound

Measuring Ascending Aortic Stiffness In Vivo in Mice Using Ultrasound

Måling Stigende Aorta Stivhed<em&gt; In Vivo</em&gt; I mus ved hjælp af ultralyd

We present a protocol for measuring in vivo aortic stiffness in mice using high-resolution ultrasound imaging. Aortic diameter is measured by ultrasound ...

Studying Pancreatic Cancer Stem Cell Characteristics for Developing New Treatment Strategies

Studere i bugspytkirtlen Kræft stamceller Karakteristik for udvikling af nye behandlingsstrategier

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor ...

A High-content In Vitro Pancreatic Islet &#946;-cell Replication Discovery Platform

A High-content In Vitro Pancreatic Islet ÃƒÅ½Ã‚Â²-cell Replication Discovery Platform

En High-indhold<em&gt; In vitro</em&gt; Pancreas Islet β-celle Replication Discovery Platform

Loss of insulin-producing &#946;-cells is a central feature of diabetes. While a variety of potential replacement therapies are being explored, expansion ...

Phosphorus-31 Magnetic Resonance Spectroscopy: A Tool for Measuring In Vivo Mitochondrial Oxidative Phosphorylation Capacity in Human Skeletal Muscle

Phosphorus-31 Magnetic Resonance Spectroscopy: A Tool for Measuring In Vivo Mitochondrial Oxidative Phosphorylation Capacity in Human Skeletal Muscle

Fosfor-31 Magnetic Resonance Spectroscopy: et redskab til måling<em&gt; In vivo</em&gt; Mitokondrie Oxidativ phosphorylering Kapacitet i human skeletmuskulatur

Skeletal muscle mitochondrial oxidative phosphorylation (OXPHOS) capacity, which is critically important in health and disease, can be measured in vivo ...

Surgical Angiogenesis in Porcine Tibial Allotransplantation: A New Large Animal Bone Vascularized Composite Allotransplantation Model

Kirurgisk angiogenese i svin Tibial Allotransplantation: en ny stor animalsk knogle vaskulariserede sammensatte Allotransplantation Model

Segmental bone loss resulting from trauma, infection malignancy and congenital anomaly remains a major reconstructive challenge. Current therapeutic ...