Name: a doxorubicin-induced murine model of dilated cardiomyopathy in vivo
Uploaded: 2020-05-16T14:00:00
Description: Watch this Scientific Journal Video about A Doxorubicin-Induced Murine Model of Dilated Cardiomyopathy In Vivo at JoVE.com

Dette arbejde blev støttet af Key Project Medical Science and Technology Development Foundation, Nanjing Department of Health (No.YKK16098).

Dyreforsøg blev godkendt af Institutional Animal Care and Use Committee (IACUC) fra Nanjing Drum Tower Hospital.
1. Tilberedning af reagenser og dyr
<ol><li>Opløs doxorubicinhydrochlorid (Pfizer, USA) i steriliseret vand. Vortex for at opnå en Dox-opløsning på 1 mg/mL og holde ved 4 °C.</li>
	<li>Brug C57BL/6 mus (8-10 uger gamle; 25-30 g vægt). Til denne undersøgelse blev mus købt fra Model Animal Research Center på Nanjing University og opbevaret i dyrerummet på Nanjing Drum Tow...

<ol>
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G., Semsarian, C., MacDonald, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dilated+cardiomyopathy.">Dilated cardiomyopathy.</a> Lancet. 390 (10092), 400-414 (2017).</li><li>Ichihara, S., 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=Attenuation+of+oxidative+stress+and+cardiac+dysfunction+by+bisoprolol+in+an+animal+model+of+dilated+cardiomyopathy.">Attenuation of oxidative stress and cardiac dysfunction by bisoprolol in an animal model of dilated cardiomyopathy.</a> Biochemical and Biophysical Research Communications. 350 (1), 105-113 (2006).</li><li>Fountoulakis, M., 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=Alterations+in+the+heart+mitochondrial+proteome+in+a+desmin+null+heart+failure+model.">Alterations in the heart mitochondrial proteome in a desmin null heart failure model.</a> Journal of Molecular and Cellular Cardiology. 38 (3), 461-474 (2005).</li><li>Fairweather, D., Rose, N. 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H., 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=A+therapeutic+anti-CD4+monoclonal+antibody+inhibits+T+cell+receptor+signal+transduction+in+mouse+autoimmune+cardiomyopathy.">A therapeutic anti-CD4 monoclonal antibody inhibits T cell receptor signal transduction in mouse autoimmune cardiomyopathy.</a> Chinese Medical Journal. 120 (15), 1319-1325 (2007).</li><li>Riad, A., 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=Toll-like+receptor-4+deficiency+attenuates+doxorubicin-induced+cardiomyopathy+in+mice.">Toll-like receptor-4 deficiency attenuates doxorubicin-induced cardiomyopathy in mice.</a> European Journal of Heart Failure. 10 (3), 233-243 (2008).</li><li>Kankeu, C., Clarke, K., Passante, E., Huber, H. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Doxorubicin-induced+chronic+dilated+cardiomyopathy-the+apoptosis+hypothesis+revisited.">Doxorubicin-induced chronic dilated cardiomyopathy-the apoptosis hypothesis revisited.</a> Journal of Molecular Medicine. 95 (3), 239-248 (2017).</li><li>Zhao, L., 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=MicroRNA-140-5p+aggravates+doxorubicin-induced+cardiotoxicity+by+promoting+myocardial+oxidative+stress+via+targeting+Nrf2+and+Sirt2.">MicroRNA-140-5p aggravates doxorubicin-induced cardiotoxicity by promoting myocardial oxidative stress via targeting Nrf2 and Sirt2.</a> Redox Biology. 15, 284-296 (2018).</li><li>O'Connell, J. L., 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=Short-term+and+long-term+models+of+doxorubicin-induced+cardiomyopathy+in+rats:+A+comparison+of+functional+and+histopathological+changes.">Short-term and long-term models of doxorubicin-induced cardiomyopathy in rats: A comparison of functional and histopathological changes.</a> Experimental and Toxicologic Pathology. 69 (4), 213-219 (2017).</li><li>Yuan, Y. P., 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=CTRP3+protected+against+doxorubicin-induced+cardiac+dysfunction,+inflammation+and+cell+death+via+activation+of+Sirt1.">CTRP3 protected against doxorubicin-induced cardiac dysfunction, inflammation and cell death via activation of Sirt1.</a> Journal of Molecular and Cellular Cardiology. 114, 38-47 (2018).</li><li>Sun, Z., 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=The+TGF-beta+pathway+mediates+doxorubicin+effects+on+cardiac+endothelial+cells.">The TGF-beta pathway mediates doxorubicin effects on cardiac endothelial cells.</a> Journal of Molecular and Cellular Cardiology. 90, 129-138 (2016).</li><li>Minotti, G., Menna, P., Salvatorelli, E., Cairo, G., Gianni, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anthracyclines:+molecular+advances+and+pharmacologic+developments+in+antitumor+activity+and+cardiotoxicity.">Anthracyclines: molecular advances and pharmacologic developments in antitumor activity and cardiotoxicity.</a> Pharmacological Reviews. 56 (2), 185-229 (2004).</li><li>Vejpongsa, P., Yeh, E. 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Dox er et uspecifikt periodisk antitumor kemoterapi lægemiddel, der almindeligvis anvendes i klinisk praksis12. Dens vigtigste bivirkning er kardiotoksicitet, karakteriseret ved kardiomyopati og efterfølgende hjertesvigt13. Den underliggende mekanisme omfatter skader på myokardie lipidperoxidation, hæmning af myokardie sarcoplasmisk reticulum Ca2+-ATPase aktivitet, og aktivering af myokardie lokale renin-angiotensin system, hvilket resulterer i øget AT II pr...

An erratum was issued for:&#160;<a href="https://www.jove.com/t/61158">A Doxorubicin-Induced Murine Model of Dilated Cardiomyopathy In Vivo</a>. A figure was updated.
Figure 1 was updated from:
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/61158/61158fig1v2.jpg" /> 
Figure 1: Schematic diagram of a Dox-induced dilated cardiomyopathy. <a href="https://www.jove.com/files/ftp_upload/61158/61158fig1v2large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
to:
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/61158/61158fig1v5.jpg" /> 
Figure 1: Schematic diagram of a Dox-induced dilated cardiomyopathy. <a href="https://www.jove.com/files/ftp_upload/61158/61158fig1v5large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>

An erratum was issued for:&#160;<a href="https://www.jove.com/t/61158">A Doxorubicin-Induced Murine Model of Dilated Cardiomyopathy In Vivo</a>. A figure was updated.

Erratum: A Doxorubicin-Induced Murine Model of Dilated Cardiomyopathy In Vivo

En af de mest almindelige årsager til hjertesvigt, DCM er karakteriseret ved ventrikulær dilatation og nedsat hjertefunktion og er den mest almindelige årsag til hjertetransplantation på verdensplan1. For yderligere at undersøge dens patogenese og finde effektive behandlinger er adgang til modne dyremodeller særlig vigtig. Formålet med de beskrevne eksperimenter er at etablere en stabil musemodel af DCM, der ligner menneskelige DCM.
På grund af DCM's komplekse patogenese er der mange forskellige metoder til at lave tilsvarende dyremodeller. Spontane DCM-modeller2 er relativt stabile, men de er dyre og ikke let tilgængelige. Genetisk modificerede dyremodeller3 er ikke veletablerede og kræver mere eksperimentel anvendelse. DCM dyremodeller forårsaget af virusinfektion4 eller autoimmune defekter5 er nemme at opnå, men de er ikke helt repræsentative for DCM. Modeller forbundet med myokardietoksicitet omfatter alkohol-induceret DCM modeller og Dox-induceret DCM dyremodeller.
Den Dox-inducerede kardiomyopatimodel opnås ved intraperitoneal injektion af Dox6. Modellen udnytter den mest alvorlige kroniske bivirkning af Dox: Efter Dox eksponering, patienter udvikler sent debut DCM symptomer med klinisk ensartethed7. Dox-induceret oxidativ stress8 og mitokondrieskader9, som fører til kardiomyocyt apoptose, er symptomer i patogenesen af DCM. Der findes akutte og kroniske Dox-behandlingsmodeller: En enkelt høj dosis Dox (15 mg/kg) fremkalder en kortsigtet model for kardiomyopati10, mens gentagne dox-injektioner med lav dosis (seks ugentlige, 3 mg/kg) fremkalder en langsigtet model for kardiomyopati11. Baseret på den præsenterede undersøgelse injiceres vilde typemus intraperitoneally en gang om ugen i en måned ved en dosis på 5 mg /kg displaymorfologi og histologi i hjertet i overensstemmelse med DCM's egenskaber ved afslutningen af behandlingen, hvilket giver en ideel måde at etablere en DCM-model på.

<table>
	<tbody>
		<tr>
			<td>4% paraformaldehyde</td>
			<td>servicebio</td>
			<td>CAS30525-89-4</td>
			<td></td>
		</tr>
		<tr>
			<td>C57BL/6 mice</td>
			<td>Model Animal Research Center of Nanjing University</td>
			<td>\</td>
			<td></td>
		</tr>
		<tr>
			<td>Doxorubicin hydrochloride</td>
			<td>Pfizer</td>
			<td>CAS25316-40-9</td>
			<td></td>
		</tr>
		<tr>
			<td>echocardiography</td>
			<td>Visualsonics</td>
			<td>\</td>
			<td></td>
		</tr>
		<tr>
			<td>Hematoxylin and Eosin staining kit</td>
			<td>Solarbio</td>
			<td>G1120</td>
			<td></td>
		</tr>
		<tr>
			<td>Masson staining kit</td>
			<td>Solarbio</td>
			<td>G1343</td>
			<td></td>
		</tr>
		<tr>
			<td>phosphate buffer solution</td>
			<td>Sigma</td>
			<td>P5368</td>
			<td></td>
		</tr>
		<tr>
			<td>potassium chloride</td>
			<td>Sigma</td>
			<td>CAS7447-40-7</td>
			<td></td>
		</tr>
		<tr>
			<td>sterilized syringe</td>
			<td>Millipore</td>
			<td>SLGP033RB</td>
			<td></td>
		</tr>
	</tbody>
</table>

a doxorubicin-induced murine model of dilated cardiomyopathy in vivo

Udvidet kardiomyopati (DCM) refererer til et spektrum af heterogene myokardiesygdomme karakteriseret ved ventrikulær dilatation og deprimeret hjertepræstation i mangel af hypertension, valvular, medfødt, eller iskæmisk hjertesygdomme, og som kan være relateret til infektion, autoimmune eller metaboliske abnormiteter, eller familie arv. Det kan udvikle sig til overbelastet hjertesvigt med en dårlig prognose. Doxorubicin (Dox) er almindeligt anvendt som et kemoterapeutisk lægemiddel, men dets anvendelse er begrænset, fordi det forårsager DCM-lignende ændringer af myokardiet. Dens myokardietoksicitet tilskrives oxidativ stress, kronisk betændelse, og kardiomyocyt apoptose. En model af DCM udnytte disse Dox-induceret DCM symptomer er ikke blevet etableret.

Hjertefunktion Udvidet kardiomyopati er karakteriseret ved progressiv ventrikulær dilatation og kontraktile dysfunktion. Figur 2 viser repræsentative ekkokardigrafiske billeder af de to grupper. Dox-behandlede mus viste markant reduceret venstre ventrikulær udslyngning fraktion og venstre ventrikulær fraktioneret afkortning (Figur 3A,B). LV-diameteren steg også i både de diastoliske og systoliske faser (<strong class="x...

Watch this Scientific Journal Video about A Doxorubicin-Induced Murine Model of Dilated Cardiomyopathy In Vivo at JoVE.com

A Doxorubicin-Induced Murine Model of Dilated Cardiomyopathy In Vivo

Beskrevet er en protokol til at etablere en Doxorubicin-induceret udvidet kardiomyopati (DCM) model i mus via langsigtet intraperitoneal injektion af Doxorubicin.

En Doxorubicin-induceret Murine Model af udvidet kardiomyopati i Vivo

Dilated cardiomyopathy (DCM) refers to a spectrum of heterogeneous myocardial disorders characterized by ventricular dilation and depressed cardiac ...

How a Doxorubicin-induced dilated cardiomyopathy model is a stable, reliable, and economical.This method will contribute to varying the pathogenesis on dilated cardiomyopathy and to exploring new drugs.Dissolve doxorubicin hydrochloride in sterilized water at a concentration of one milligram per milliliter.Vortex the solution and store it at four degrees Celsius.Maintain eight to 10-week-old mice in pathogen-free mouse cages with a 12-hour light-dark cycle at a constant temperature of 23 degrees Celsius.And provide as much food and water as needed.Before beginning treatment, randomize mice into a control group and a dox group.For mice in the dox group, use one milliliter sterilized syringes to inject the dox solution intraperitoneally.Administer a dose of five milligrams per kilogram of body weight intraperitoneally once per week.Perform the injection in the lower left or right quadrant of the abdomen, taking care of to avoid the urinary bladder and other abdominal organs.For four weeks measure the body weight of the mice weekly and adjust the injection dose accordingly each week.After confirming anesthesia, use depilatory cream to carefully remove the chest fur.After performing echocardiography, use the M-mode tracings to measure the average LV end

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