Name: a doxorubicin-induced murine model of dilated cardiomyopathy in vivo
Uploaded: 2020-05-16T14:00:00.000+00:00
Duration: 5 min 14 s
Description: Watch this Scientific Journal Video about A Doxorubicin-Induced Murine Model of Dilated Cardiomyopathy In Vivo at JoVE.com

この研究は、南京省保健局(No.YKK16098)の主要プロジェクト医学技術開発財団によって支援されました。

動物実験は、南京ドラムタワー病院の施設動物のケアと使用委員会(IACUC)によって承認されました。
試薬及び動物の調製
<ol><li>ドキソルビシン塩酸塩(ファイザー、米国)を殺菌水に溶かします。ボルテックスは、1 mg/mLのドークス溶液を得て、4°Cに保つ。</li>
	<li>C57BL/6マウス(8~10週齢、体重25~30g)を使用してください。この研究のために、マウスは南京大学モデル動物研究センターから購入し、南京ドラムタワー病院の動物室に保管されました。</li>
	<li>病原体を含まないマウスケージは、23°Cの一定温度で12時間の明暗サイクル下で維持した。 すべての動物は通常のチャウダイエットで餌を与えられ、食べ物と水のアドリビタムを得ました。</li>
</ol>2. DCM動物モデルの確立
<ol><li>マウスを正常(n=5)群とDox(n=5)群に無作為化する。</li>
	<li>Dox群に対して週1回1mL滅菌シリンジを用いて、5mg/kgの用量で腹腔内溶液を投与する。同量の生理液で同じ方法でコントロールマウスを治療する。</li>
	<li>週に2つのグループの体重を測定します。したがって、体重に基づいて毎週、合計4週間、累積用量20mg/kgを調整します(図1)。 注:4週目の心エコー検査は、2つのグループ間の心機能に有意な差を示したので、4週間の期間が選択されました。</li>
</ol>3. 心エコー検査
<ol><li>第4週の終わりに、マウスの心エコー検査を行う。</li>
	<li>マウスを2%イオブルランを鼻腔内に麻酔します。マウスが歯付きピンシーザーで皮膚のピンチに反応しない場合、またはつま先と尾を刺激する場合は、プロトコルを続行します。</li>
	<li>動物の取り扱いプラットフォーム上のマウスを、サピーヌ位置に置きます。麻酔を維持するには、動物の鼻と口を鼻コーンで覆い、2%のイオブルランを送達する。 注:IP麻酔の場合は、0.2 mL/20 gの用量で4%塩素水和物を注入してください。</li>
	<li>電動の剃り機で胸の毛皮を慎重に取り外します。胸部心エコー検査を用いて生体内の心機能を評価する。</li>
	<li>233 Hzのフレームレートで、パラスターナル長軸ビューと短軸ビューの両方でLV心エコー図を行います。</li>
	<li>Mモードトレースでは、平均LV末端収縮期直径(LVIDd)、LV末端拡張期直径(LVID)、心室内隔壁厚(IVS)、LV後壁厚(LVPW)を3~5回の心臓拍から測定します。また、心エコー検査に基づいて、放出分数(EF)と分数短縮(FS)も計算します。</li>
</ol>4. 組織学的染色
<ol><li>心エコー分析の後、10%塩化カリウムの心室内注射によりマウスを犠牲にする。</li>
	<li>肝臓と肺が青白くなるまで、解剖後に約30mLの生理液で心臓を浸透する。</li>
	<li>心臓を切除し、血液を押し出すためにリン酸塩緩衝液で十分に洗浄します。</li>
	<li>24時間室温で4%ホルマリンで心臓を固定し、パラフィンワックスが冷却して固化するようにパラフィンボックスで組織を処理します。</li>
	<li>心臓を5μmの厚さのスライスに切り、病理学的染色を行います。</li>
	<li>乳頭筋を含む脱ワックスおよび水分補給の切片。<ol>
<li>55°Cで30分間のインキュベートスライド。次に、キシレン2xで2分ずつインキュベートします。100%エタノール2x各2分間;95% エタノール 2 x 各 2 分;2分間の80%エタノール;75% エタノール 2 分間;50%エタノールを2分間使用します。</li>
	</ol>
</li>
	<li>ヘマトキシリンとエオシン(H&E)とマッソンの汚れを使用したステイン。</li>
</ol>

<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. 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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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利益相反は宣言されていません。

Doxは、臨床実践12で一般的に使用される非特異的周期的な抗腫瘍性化学療法薬である。その主な副作用は、心筋症とその後の心不全13によって特徴付けられる心毒性である。基になる機構は、心筋脂質過酸化の損傷、心筋サルコプラスミックリチラムCa2+-ATPase活性の阻害、および心筋局所レニンアンジオテンシン系の活性化を含み、AT II産生および細...

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

心不全の最も一般的な原因の1つは、DCMは心室拡張と心機能の低下を特徴とし、世界中の心臓移植の最も一般的な理由である1。その病因をさらに調査し、効果的な治療法を見つけるためには、成熟した動物モデルへのアクセスが特に重要です。本記載の実験の目的は、ヒトDCMに似たDCMの安定したマウスモデルを確立することにある。
DCMの複雑な病因により、対応する動物モデルを作るための多くの異なる方法があります。自発的なDCMモデル2 は比較的安定していますが、高価で簡単には利用できません。遺伝子組み換え動物モデル3 は確立されておらず、より実験的な使用が必要である。ウイルス感染4 または自己免疫欠損5 によって誘導されるDCM動物モデルは容易に得られるが、完全にDCMを代表するわけではない。心筋毒性に関連するモデルには、アルコール誘導DCMモデルおよびDox誘発DCM動物モデルが含まれる。
Dox誘発型心筋症モデルは、Dox6の腹腔内注射により得られる。このモデルは、Doxの最も重篤な慢性副作用を利用する:Dox暴露後、患者は臨床均一性を有する後期発症DCM症状を発症する7.心筋細胞アポトーシスを引き起こすドックス誘導酸化ストレス8 およびミトコンドリア損傷9は、DCMの病因における症状である。急性および慢性のDox治療モデルがあります:単一の高用量のDox(15mg/kg)は心筋症の短期モデルを誘発します10,繰り返し低用量のDox注射 (6 週, 3 mg/kg) 心筋症の長期モデルを誘導します。本研究に基づき、野生型マウスは週に1回、1ヶ月間、5mg/kgの用量で腹腔内注射を行い、治療の終了までにDCMの特性と一致する心臓の形態および組織学を表示し、DCMモデルを確立するための理想的な方法を提供する。

<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

拡張型心筋症(DCM)は、高血圧、弁膜、先天性、または虚血性心疾患がない場合の心室拡張およびうつ心臓のパフォーマンスを特徴とする異種心筋障害のスペクトルを指し、感染、自己免疫性または代謝異常、または家族の遺伝に関連する可能性がある。予後が悪いとうっ血性心不全に進行する可能性があります。ドキソルビシン(Dox)は化学療法薬として広く採用されているが、心筋のDCM様変化を引き起こすのでその使用は限られている。その心筋毒性は、酸化ストレス、慢性炎症、心筋細胞アポトーシスに起因する。これらのDox誘発DCM症状を利用するDCMのモデルは確立されていない。

心臓機能 拡張型心筋症は、進行性心室拡張および収縮機能障害を特徴とする。 図2 は、2つのグループの代表的な心エコー画像を示す。ドックス処理マウスは、左心室駆出率および左心室分画短縮を著しく減少させた(図3A,B)。また、LV径は拡張期および収縮期の両方で増加した(図3C,D)。...

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

記載は、ドキソルビシンの長期腹腔内注射を介してマウスにおけるドキソルビシン誘発拡張型心筋症(DCM)モデルを確立するためのプロトコルである。

インビボにおける拡張型心筋症のドキソルビシン誘発性マウスモデル

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

a-doxorubicin-induced-murine-model-of-dilated-cardiomyopathy-in-vivo

Research

JoVE Journal

Medicine

4.4K ビュー.  Clinical College of Nanjing Medical University. 記載は、ドキソルビシンの長期腹腔内注射を介してマウスにおけるドキソルビシン誘発拡張型心筋症(DCM)モデルを確立するためのプロトコルである。

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

Displacement Analysis of Myocardial Mechanical Deformation (DIAMOND) Reveals Segmental Heterogeneity of Cardiac Function in Embryonic Zebrafish

Zebrafish are increasingly utilized as a model organism for cardiomyopathies and regeneration. Current methods evaluating cardiac function fail to reliably detect segmental mechanics and are not readily feasible in zebrafish. Here we present a semiautomated, open-source method for the quantitative assessment of four-dimensional (4D) segmental cardiac function: displacement analysis of myocardial mechanical deformation (DIAMOND). Transgenic embryonic zebrafish were imaged in vivo using a light-sheet fluorescence microscopy system with 4D cardiac motion synchronization. Acquired 3D digital hearts were reconstructed at end-systole and end-diastole, and the ventricle was manually segmented into binary datasets. Then, the heart was reoriented and isotropically resampled along the true short axis, and the ventricle was evenly divided into eight portions (I&#8211;VIII) along the short axis. Due to the different resampling planes and matrices at end-systole and end-diastole, a transformation matrix was applied for image registration to restore the original spatial relationship between the resampled systolic and diastolic image matrices. After image registration, the displacement vector of each segment from end-systole to end-diastole was calculated based on the displacement of mass centroids in three dimensions (3D). DIAMOND shows that basal myocardial segments adjacent to the atrioventricular canal undergo the highest mechanical deformation and are the most susceptible to doxorubicin-induced cardiac injury. Overall, DIAMOND provides novel insights into segmental cardiac mechanics in zebrafish embryos beyond traditional ejection fraction (EF) under both physiological and pathological conditions.

Displacement Analysis of Myocardial Mechanical Deformation DIAMOND Reveals Segmental Heterogeneity of Cardiac Function in Embryonic Zebrafish

The goal of this protocol is to detail a novel method for the assessment of segmental cardiac function in embryonic zebrafish under both physiological and pathological conditions.

心筋機械変形(DIAMOND)の変位解析は、胚性ゼブラフィッシュにおける心機能のセグメント的不均一性を明らかにする

Zebrafish are increasingly utilized as a model organism for cardiomyopathies and regeneration. Current methods evaluating cardiac function fail to ...

発生生物学

Permanent Ligation of the Left Anterior Descending Coronary Artery in Mice: A Model of Post-myocardial Infarction Remodelling and Heart Failure

Heart failure is a syndrome in which the heart fails to pump blood at a rate commensurate with cellular oxygen requirements at rest or during stress. It is characterized by fluid retention, shortness of breath, and fatigue, in particular on exertion. Heart failure is a growing public health problem, the leading cause of hospitalization, and a major cause of mortality. Ischemic heart disease is the main cause of heart failure.
Ventricular remodelling refers to changes in structure, size, and shape of the left ventricle. This architectural remodelling of the left ventricle is induced by injury (e.g., myocardial infarction), by pressure overload (e.g., systemic arterial hypertension or aortic stenosis), or by volume overload. Since ventricular remodelling affects wall stress, it has a profound impact on cardiac function and on the development of heart failure. A model of permanent ligation of the left anterior descending coronary artery in mice is used to investigate ventricular remodelling and cardiac function post-myocardial infarction. This model is fundamentally different in terms of objectives and pathophysiological relevance compared to the model of transient ligation of the left anterior descending coronary artery. In this latter model of ischemia/reperfusion injury, the initial extent of the infarct may be modulated by factors that affect myocardial salvage following reperfusion. In contrast, the infarct area at 24 hr after permanent ligation of the left anterior descending coronary artery is fixed. Cardiac function in this model will be affected by 1) the process of infarct expansion, infarct healing, and scar formation; and 2) the concomitant development of left ventricular dilatation, cardiac hypertrophy, and ventricular remodelling.
Besides the model of permanent ligation of the left anterior descending coronary artery, the technique of invasive hemodynamic measurements in mice is presented in detail.

Heart failure is the leading cause of hospitalization and a major cause of mortality. A model of permanent ligation of the left anterior descending coronary artery in mice is applied to investigate ventricular remodelling and cardiac dysfunction post-myocardial infarction. The technique of invasive hemodynamic measurements in mice is presented.

マウスの左前下冠動脈の動脈の常設ライゲーション：心筋梗塞後のリモデリングおよび心不全のモデル

Heart failure is a syndrome in which the heart fails to pump blood at a rate commensurate with cellular oxygen requirements at rest or during stress. It ...

医学

Preclinical Cardiac Electrophysiology Assessment by Dual Voltage and Calcium Optical Mapping of Human Organotypic Cardiac Slices

Human cardiac slice preparations have recently been developed as a platform for human physiology studies and therapy testing to bridge the gap between animal and clinical trials. Numerous animal and cell models have been used to examine the effects of drugs, yet these responses often differ in humans. Human cardiac slices offer an advantage for drug testing in that they are directly derived from viable human hearts. In addition to having preserved multicellular structures, cell-cell coupling, and extracellular matrix environments, human cardiac tissue slices can be used to directly test the effect of innumerable drugs on adult human cardiac physiology. What distinguishes this model from other heart preparations, such as whole hearts or wedges, is that slices can be subjected to longer-term culture. As such, cardiac slices allow for studying the acute as well as chronic effects of drugs. Furthermore, the ability to collect several hundred to a thousand slices from a single heart makes this a high-throughput model to test several drugs at varying concentrations and combinations with other drugs at the same time. Slices can be prepared from any given region of the heart. In this protocol, we describe the preparation of left ventricular slices by isolating tissue cubes from the left ventricular free wall and sectioning them into slices using a high precision vibrating microtome. These slices can then either be subjected to acute experiments to measure baseline cardiac electrophysiological function or cultured for chronic drug studies. This protocol also describes dual optical mapping of cardiac slices for simultaneous recordings of transmembrane potentials and intracellular calcium dynamics to determine the effects of the drugs being investigated.

This protocol describes the procedure for sectioning and culturing human cardiac slices for preclinical drug testing and details the use of optical mapping for recording transmembrane voltage and intracellular calcium signals simultaneously from these slices.

ヒトの神経ノチカルスライスの二重電圧およびカルシウム光学マッピングによる前臨床心臓電気生理学的評価

Human cardiac slice preparations have recently been developed as a platform for human physiology studies and therapy testing to bridge the gap between ...

生物工学

Percutaneous Contrast Echocardiography-guided Intramyocardial Injection and Cell Delivery in a Large Preclinical Model

Cell and gene therapy are exciting and promising strategies for the purpose of cardiac regeneration in the setting of heart failure with reduced ejection fraction (HFrEF). Before they can be considered for use, and implemented in humans, extensive preclinical studies are required in large animal models to evaluate the safety, efficacy, and fate of the injectate (e.g., stem cells) once delivered into the myocardium. Small rodent models offer advantages (e.g., cost effectiveness, amenability for genetic manipulation); however, given inherent limitations of these models, the findings in these rarely translate into the clinic. Conversely, large animal models such as rabbits, have advantages (e.g., similar cardiac electrophysiology compared to humans and other large animals), whilst retaining a good cost-effective balance. Here, we demonstrate how to perform a percutaneous contrast echocardiography-guided intramyocardial injection (IMI) technique, which is minimally invasive, safe, well tolerated, and very effective in the targeted delivery of injectates, including cells, into several locations within the myocardium of a rabbit model. For the implementation of this technique, we also have taken advantage of a widely available clinical echocardiography system. After putting in practice the protocol described here, a researcher with basic ultrasound knowledge will become competent in the performance of this versatile and minimally invasive technique for routine use in experiments, aimed at hypothesis testing of the capabilities of cardiac regenerative therapeutics in the rabbit model. Once competency is achieved, the whole procedure can be performed within 25 min after anaesthetizing the rabbit.

Novel therapeutic strategies in cardiac regenerative medicine require extensive and detailed studies in large preclinical animal models before they can be considered for use in humans. Here, we demonstrate a percutaneous contrast echocardiography-guided intramyocardial injection technique in rabbits, which is valuable for hypothesis testing the efficacy of such novel therapies.

経皮的造影エコー ガイド心筋と大規模な前臨床モデルで携帯配信

Cell and gene therapy are exciting and promising strategies for the purpose of cardiac regeneration in the setting of heart failure with reduced ejection ...

Tachycardia-Induced Cardiomyopathy As a Chronic Heart Failure Model in Swine

A stable and reliable model of chronic heart failure is required for many experiments to understand hemodynamics or to test effects of new treatment methods. Here, we present such a model by tachycardia-induced cardiomyopathy, which can be produced by rapid cardiac pacing in swine.
A single pacing lead is introduced transvenously into fully anaesthetized healthy swine, to the apex of the right ventricle, and fixated. Its other end is then tunneled dorsally to the paravertebral region. There, it is connected to an in-house modified heart pacemaker unit that is then implanted in a subcutaneous pocket. 
After 4 - 8 weeks of rapid ventricular pacing at rates of 200 - 240 beats/min, physical examination revealed signs of severe heart failure - tachypnea, spontaneous sinus tachycardia, and fatigue. Echocardiography and X-ray showed dilation of all heart chambers, effusions, and severe systolic dysfunction. These findings correspond well to decompensated dilated cardiomyopathy and are also preserved after the cessation of pacing.
This model of tachycardia-induced cardiomyopathy can be used for studying the pathophysiology of progressive chronic heart failure, especially hemodynamic changes caused by new treatment modalities like mechanical circulatory supports. This methodology is easy to perform and the results are robust and reproducible.

Here, we present a protocol to produce tachycardia-induced cardiomyopathy in swine. This model represents a potent way to study the hemodynamics of progressive chronic heart failure and the effects of applied treatment.

豚の慢性心不全モデルとして頻拍誘発性心筋

A stable and reliable model of chronic heart failure is required for many experiments to understand hemodynamics or to test effects of new treatment ...

A Doxorubicin-induced Cardiomyopathy Model in Adult Zebrafish

The genetically accessible adult zebrafish (Danio rerio) has been increasingly used as a vertebrate model for understanding human diseases such as cardiomyopathy. Because of its convenience and amenability to high throughput genetic manipulations, the generation of acquired cardiomyopathy models, such as the doxorubicin-induced cardiomyopathy (DIC) model in adult zebrafish, is opening the doors to new research avenues, including discovering cardiomyopathy modifiers via forward genetic screening. Different from the embryonic zebrafish DIC model, both initial acute and later chronic phases of cardiomyopathy can be determined in the adult zebrafish DIC model, enabling the study of stage-dependent signaling mechanisms and therapeutic strategies. However, variable results can be obtained with the current model, even in the hands of experienced investigators. To facilitate future implementation of the DIC model, we present a detailed protocol on how to generate this DIC model in adult zebrafish and describe two alternative ways of intraperitoneal (IP) injection. We further discuss options on how to reduce variations to obtain reliable results and provide suggestions on how to appropriately interpret the results.

A method to generate a doxorubicin-induced cardiomyopathy model in adult zebrafish (Danio rerio) is described here. Two alternative ways of intraperitoneal injection are presented and conditions to reduce variations among different experimental groups are discussed.

大人のゼブラフィッシュのドキソルビシン誘発心筋症モデル

The genetically accessible adult zebrafish (Danio rerio) has been increasingly used as a vertebrate model for understanding human diseases such as ...

Murine Myocardial Infarction Model using Permanent Ligation of Left Anterior Descending Coronary Artery

Myocardial infarction (MI) and acute coronary diseases are among the most prominent causes of death in population with western lifestyle. The murine models of MI with permanent ligation of left-anterior descending (LAD) coronary artery closely mimics MI in humans. Murine models benefit from the extensive genetic engineering available nowadays. Here we propose a reproducible murine surgical model of myocardial infarction by permanent LAD coronary ligation. Our technique comprises anesthesia with ketamine/xylazine that can be rapidly reversed by administration of an antagonist, intubation without tracheotomy for mechanical-assisted ventilation, ventilation with application of extrinsic positive end-expiratory pressure (PEEP) to avoid alveolar collapse, a thoracotomy method limiting to the minimum surgical lesions made to skeletal muscles, and lung inflation without thoracentesis. This method is sparsely invasive, reproducible and reduces post-surgery mortality and complications.

Herein we describe a surgical procedure showing how to achieve permanent ligation of the left-anterior descending coronary artery in mice. This model is of high relevance to investigate the pathophysiology of myocardial infarction and the concomitant biological processes.

左前皮下冠動脈の永久的なライゲーションを用いたマウス心筋梗塞モデル

Myocardial infarction (MI) and acute coronary diseases are among the most prominent causes of death in population with western lifestyle. The murine ...

Revealing Subtle Changes in Cardiac Function using Transthoracic Dobutamine Stress Echocardiography in Mice

Left ventricular (LV) dysfunction paves the final pathway for a multitude of cardiac disorders. With the non-invasive high-frequency transthoracic dobutamine stress echocardiography in humans, a reductionist investigation approach to unmask subtle changes in cardiac function has become possible. Here, we provide a protocol for using this technique in mice to facilitate expanded analysis of LV architecture and function in physiology and pathology enabling the observation of alterations in models of cardiac disease hidden in unstressed hearts. This investigation can be performed in one and the same animal and allows both, basal and pharmacologically stress-induced measurements. We outline detailed criteria for appropriate anesthesia, imaging-based LV analysis, consideration of intra- and interobserver variability, and obtaining positive inotrope response that can be attained in mice after intraperitoneal injection of dobutamine under near physiological conditions. To recapitulate the characteristics of human physiology and disease in small animal models, we highlight critical pitfalls in evaluation, e.g., a pronounced Bowditch effect in mice. To further meet translational objectives, we compare stress-induced effects in humans and mice. When used in translational studies, attention must be paid to physiological differences between mice and human. Experimental rigor dictates that some parameters assessed in patients can only be used with caution due to restrictions in spatial and temporal resolution in mouse models.

Left ventricular dysfunction constitutes the final common pathway for a host of cardiac disorders. We here provide a detailed protocol of transthoracic dobutamine stress echocardiography approach for comprehensive evaluation of the left ventricular function of mouse models of cardiac disease as well as cardiac phenotyping.

Left ventricular (LV) dysfunction paves the final pathway for a multitude of cardiac disorders. With the non-invasive high-frequency transthoracic ...

An Experimental Model of Myocardial Infarction for Studying Cardiac Repair and Remodeling in Knockout Mice

Cardiovascular disease is the most prevalent cause of death in Western countries, with acute myocardial infarction (MI) being the most prevalent form. This paper describes a protocol for studying the role of galectin 3 (Gal-3) in the temporal evolution of cardiac healing and remodeling in an experimental animal model of MI. 
The procedures described include an experimental model of MI with a permanent coronary ligature in male C57BL/6J (control) and Gal-3 knockout (KO) mice, an echocardiography procedure to study cardiac remodeling and systolic function in vivo, a histological evaluation of interstitial myocardial fibrosis with picrosirius red-stained and rhodamine-conjugated lectin-stained sections for studying myocyte hypertrophy by the cross-sectional area (MCSA), and the quantification of infarct size and cardiac remodeling (scar thinning, septum thickness, and expansion index) by planimetry in slices stained with Masson's trichrome and triphenyl tetrazolium chloride. Gal-3 KO mice with MI showed disrupted cardiac remodeling and an increase in the scar thinning ratio and the expansion index. At the onset of MI, myocardial function and cardiac remodeling were also severely affected. At 4 weeks post MI, the natural evolution of fibrosis in infarcted Gal-3 KO mice was also affected. 
In summary, the experimental model of MI is a suitable model for studying the temporal evolution of cardiac repair and remodeling in mice with the genetic deletion of Gal-3 and other animal models. The lack of Gal-3 affects the dynamics of cardiac repair and disrupts the evolution of cardiac remodeling and function after MI.

Here, we describe an experimental model of myocardial infarction, an echocardiography procedure to study cardiac remodeling and function, and procedures for quantifying fibrosis and hypertrophy in picrosirius red-stained and rhodamine-stained sections, as well as the infarct size and expansion index in slices stained with Masson's trichrome.

Cardiovascular disease is the most prevalent cause of death in Western countries, with acute myocardial infarction (MI) being the most prevalent form. ...

<meta charset="utf8"></head><body>高脂肪食とSTZ注射による糖尿病性心筋症のマウスモデル

Modeling and Evaluation of Murine Diabetic Cardiomyopathy Model

The underlying pathophysiological mechanisms of diabetic cardiomyopathy (DbCM), a leading cause of mortality among patients with type 2 diabetes mellitus (T2DM), remain poorly understood. The myocardial toxicity associated with T2DM is attributed to factors such as lipotoxicity, glucotoxicity, oxidative stress, reduced cardiac efficiency, and lipoapoptosis. Compared to rats, mice offer greater accessibility, cost-effectiveness, and broader applicability for animal experiments. Insulin resistance and impaired insulin secretion are crucial factors in the pathophysiology of T2DM. We introduce a novel nongenetic murine model that replicates the progression of human DbCM induced by a combination of high-fat diet (HFD) feeding and streptozotocin (STZ) injection. In this study, we used wild-type C57BL/6J mice, administering an HFD regimen for 12 weeks, followed by intraperitoneal injections of STZ for an additional 12 weeks to induce characteristic manifestations of T2DM. We conducted oral glucose tolerance tests and measured serum insulin concentrations to confirm the development of insulin resistance and insufficient insulin secretion. Cardiac structure and function were rigorously assessed through noninvasive transthoracic echocardiography. Pathological characteristics were evaluated through Masson's trichrome staining and wheat germ agglutinin (WGA) staining, revealing pathological features related to DbCM. Therefore, we provide a robust and versatile method for establishing a nongenetic murine model of DbCM.

<meta charset="utf8"></head><body>マウス糖尿病性心筋症モデルのモデリングと評価

This protocol outlines a method for inducing diabetic cardiomyopathy through a combination of high-fat diet feeding and streptozotocin injection. This approach aims to provide a reliable framework for scientific investigation into diabetic cardiomyopathy and to explore potential avenues for clinical treatment applications.

マウス糖尿病性心筋症モデルのモデリングと評価

The underlying pathophysiological mechanisms of diabetic cardiomyopathy (DbCM), a leading cause of mortality among patients with type 2 diabetes mellitus ...

インビボにおける拡張型心筋症のドキソルビシン誘発性マウスモデル

要約

さらに動画を探す

この動画の章

心筋機械変形(DIAMOND)の変位解析は、胚性ゼブラフィッシュにおける心機能のセグメント的不均一性を明らかにする

マウスの左前下冠動脈の動脈の常設ライゲーション：心筋梗塞後のリモデリングおよび心不全のモデル

ヒトの神経ノチカルスライスの二重電圧およびカルシウム光学マッピングによる前臨床心臓電気生理学的評価

経皮的造影エコーガイド心筋と大規模な前臨床モデルで携帯配信

豚の慢性心不全モデルとして頻拍誘発性心筋

大人のゼブラフィッシュのドキソルビシン誘発心筋症モデル

左前皮下冠動脈の永久的なライゲーションを用いたマウス心筋梗塞モデル

Revealing Subtle Changes in Cardiac Function using Transthoracic Dobutamine Stress Echocardiography in Mice

An Experimental Model of Myocardial Infarction for Studying Cardiac Repair and Remodeling in Knockout Mice

マウス糖尿病性心筋症モデルのモデリングと評価

インビボにおける拡張型心筋症のドキソルビシン誘発性マウスモデル

要約

さらに動画を探す

この動画の章

心筋機械変形(DIAMOND)の変位解析は、胚性ゼブラフィッシュにおける心機能のセグメント的不均一性を明らかにする

マウスの左前下冠動脈の動脈の常設ライゲーション：心筋梗塞後のリモデリングおよび心不全のモデル

ヒトの神経ノチカルスライスの二重電圧およびカルシウム光学マッピングによる前臨床心臓電気生理学的評価

経皮的造影エコー ガイド心筋と大規模な前臨床モデルで携帯配信

豚の慢性心不全モデルとして頻拍誘発性心筋

大人のゼブラフィッシュのドキソルビシン誘発心筋症モデル

左前皮下冠動脈の永久的なライゲーションを用いたマウス心筋梗塞モデル

Revealing Subtle Changes in Cardiac Function using Transthoracic Dobutamine Stress Echocardiography in Mice

An Experimental Model of Myocardial Infarction for Studying Cardiac Repair and Remodeling in Knockout Mice

マウス糖尿病性心筋症モデルのモデリングと評価

経皮的造影エコーガイド心筋と大規模な前臨床モデルで携帯配信