The authors' work was supported by a grant from Ministry of Science and Technology (MOST 103-2320-B-002-068-MY2), the National Health Research Institute (NHRI-EX104-10418SC), and National Taiwan University (NTU 104R4000).

全ての動物実験は、実験動物の管理と使用に関する指針に従って行わ米国国立衛生研究所によって公開（NIH公開番号85-23、1996年改訂）しました。このプロトコルはにより、国立台湾大学の施設内動物管理使用委員会によって定められたガイドラインに基づいて承認されました。 1.動物の手術<ol><li>ホーニング石の上に針の先端を鈍化により、22 G注射針を準備します。ペンチを使用して、直角に針をひだのあります。 </li><li>手術前に、必要な手術器具や材料、ならびに回復ケージを準備します。使用前にすべての機器および外科用品をオートクレーブ。 </li><li> 200グラムの周りにラットを維持し、食物および水に自由にアクセス制御された温度（21±2℃）で12時間の明/暗サイクルの下でそれらを保ちます。各グループに少なくとも6匹のラットが含まれます。ペントバルビタールでラットを麻酔または別の適切な麻酔薬（0.5ミリリットル、腹腔内を75mg / kg）を。尾の反射をテストすることによって、麻酔の深さを確認してください。 注：テール反射の欠如は、適切な麻酔の指標です。 </li><li>体温を維持するために加熱パッドで手術台に仰臥位にラットを置きます。外科的な汚染を避けるために、動物のバリカンヘアリムーバーローションでラットの腹部を剃ります。手術前にベタジンまたは別のクレンジング試薬ときれい-坊主腹部をスクラブ。 注：これは、作業中は無菌領域を維持することが重要です。 </li><li>メスで、腹部の正中線に沿って2cmの切開を行います。通常の生理食塩水を使用して、手術中に湿った腹部臓器を維持します。後部腹膜領域にある下大静脈を露出させるためにコットンボールを使用して、側に慎重に消化器官を置き換えます。並置され、通常は上にある腹部大動脈を、識別下大静脈の左。 注：腹部大動脈は、心拍数に合わせて脈動する容器です。 </li><li>下の血管を明らかにピンセットで腹膜を貫通。静かに腎動脈に隣接する腹部大動脈を隔離し、右の起源と左腎動脈との間に腹部大動脈の下に8センチメートル長4-0絹縫合糸を渡します。 </li><li>縫合糸と緩い二重結び目を作ります。直径3mmのループを残して、ループ内の鈍化と曲がっ22 G針を配置します。大動脈と針の周りに結び目を締めて、直径0.7mmの収縮を達成するために、直ちに針を取り除きます。 </li><li> 6-0吸収性縫合糸材料と腹腔を閉じます。単純結節縫合で筋肉や皮膚切開を縫合。感染を防ぐために、ヨードチンキで手術部位を洗浄。 </li><li>それは十分な意識を取り戻すまで、自由な移動によって示されるように、慎重に動物を観察しますおよび食物摂取。それは胸骨横臥位を維持するのに十分な意識を取り戻したまで無人の動物を放置しないでください。術後の痛みを防ぐために、アセトアミノフェン（0.5ミリリットル中300ミリグラム/ kg、腹腔内）でラットを扱います。 注：制度的政策によって承認されたように、ユーザは、鎮痛薬を投与すべきです。 </li></ol> 2.組織および血液サンプル採取<ol><li> 10週間後の手術では、ラットを計量し、ペントバルビタール（0.5ミリリットル、腹腔内を75mg / kg）で、それを麻酔。手術前に、その尾の反射をテストすることによって、麻酔の深さを確認します。金属製のトレイにラットを置きます。 </li><li>メスを用いて首の正中線に沿って2cmの切開を行います。気管を露出するためにピンセットで慎重に筋肉を変位させます。気管に平行であり、心拍数と時間的に脈動頚動脈を識別するために慎重に観察します。 </li><li> EDTAでコーティングされた採血管に頸動脈から血液を採取します。すぐに遠心分離機2,000×gで15分間、血液および血漿を収集します。必要になるまで-80℃で血漿を保管してください。 </li><li>剣状突起の正中線の周りに、胸部に5cmの切開を行います。鋭いピンセットでダイアフラムを突き刺します。ハサミを使用して、心臓を露出するために、両側の半ば鎖骨ラインに沿って肋骨を切断して除去します。心臓と血管の境界線に沿って慎重に心を切り出します。組織をつかんずに静かに心を削除します。 </li><li>灌流針に大動脈トランクを結びつけることによって修正されたランゲンドルフ灌流装置に心をマウントします。クレブス緩衝液で心臓灌流（110 mMの塩化ナトリウム、2.6のKClを含有する1.2mMのKH 2 PO 4、1.2mMのMgSO 4を、25mMのNaHCO 3を、および11 mMグルコース[pHが7.4]）は、血液を洗い流します。心を計量し、心臓重量対体重比を算出します。氷上で4％パラホルムアルデヒドで心臓を修正しました。パラホルムアルデヒド蒸気はTOXであるため、マスクを着用することを忘れないでくださいIC。 </li></ol> 3.組織線維症の定量化<ol><li>組織切片デバイス上でパラホルムアルデヒド固定心臓組織を置き、2ミリメートルの厚さの切片を切りました。包埋カセットに組織切片を置きます。段階的なアルコール浴一連の組織を脱水（50％、75％、95％、1時間毎、100％）。 </li><li> 1時間ワックスに最終的に1時間キシレンで組織に浸潤し。包埋カセットに浸透組織を置き、パラフィンワックスに埋め込むことができます。ミクロトームまで室温でパラフィンブロック中に包埋組織を保管してください。 </li><li>厚さ4μmの切片に埋め込まれた組織をスライス。 45ºCの水浴中でセクションを配置します。角度で水浴中にスライドガラスを浸し、徐々にスライドへの部分的な付着を可能にするためにパラフィン切片のエッジに近づきます。 </li><li>組織切片の下で潜在的なエアポケットを除去するために、より良い取り付けを容易にするために、お風呂の中と外にスライドを移動します。ドライトン彼は1時間37ºCでスライドし、組織染色、室温で保管してください。 </li><li>タンクにスライドを置きます。 30分間キシレンでスライドを脱パラフィン化し、最後に蒸留水で（3分間それぞれ95％、75％、50％）を順次希釈アルコールでそれを再水和します。完全に1時間組織切片をカバーするために十分なピクロシリウス赤色の溶液を使用してください。 2変更のための0.5％酢酸溶液中でスライドを洗浄し、その後無水アルコールで2回のすすぎを行います。 </li><li>スライドを空気乾燥し、カバースリップで合成樹脂でスライドをマウントします。可視光フィールドでスライドを撮影。 注：写真の中の赤い部分は、200Xの倍率で顕微鏡下ピクロシリウス赤のプラスゾーンを示しています。線維症11の程度を示す総面積、オーバーピクロシリウス赤のプラスゾーンの割合を計算します。 </li></ol> 4.ブラッドトロポニン定量<ol><li>血漿トロポニンレベルを定量酵素結合免疫吸着アッセイ（ELISA）を使用。重複の各サンプルをアッセイ。負荷血漿50μlの、適切なウェルに抗体カクテル50μlの。プレートを密封し、400rpmでプレートシェーカー上で室温で1時間インキュベートします。 注：血漿中の心臓トロポニンは、心臓傷害のマーカーです。 </li><li>液体を吸引し、洗浄緩衝液250μlの3回各ウェルを洗浄します。最後の洗浄後、プレートを反転し、過剰な液体を除去するために清潔なペーパータオルに対してそれをブロット。 </li><li>各ウェルにテトラメチルベンジジン基質100μlを加え、400rpmでプレートシェーカー上で暗所で10分間インキュベートします。各ウェルに停止液100μLを加えます。混合するために1分間プレートシェーカー上でプレートを振ります。 450nmでの光学密度（OD）を記録します。 注：トロポニン濃度がOD値に比例します。 </li></ol>

<ol>
	<li>Houser, S. R., 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=Animal+models+of+heart+failure:+a+scientific+statement+from+the+American+Heart+Association.">Animal models of heart failure: a scientific statement from the American Heart Association.</a> Circ Res. 111, 131-150 (2012).</li><li>Towbin, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inherited+cardiomyopathies.">Inherited cardiomyopathies.</a> Circ J. 78, 2347-2356 (2014).</li><li>Breckenridge, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heart+failure+and+mouse+models.">Heart failure and mouse models.</a> Dis Model Mech. 3, 138-143 (2010).</li><li>van Bilsen, M., van Nieuwenhoven, F. A., van der Vusse, G. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metabolic+remodelling+of+the+failing+heart:+beneficial+or+detrimental?.">Metabolic remodelling of the failing heart: beneficial or detrimental?.</a> Cardiovasc Res. 81, 420-428 (2009).</li><li>Patten, R. D., Hall-Porter, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Small+animal+models+of+heart+failure:+development+of+novel+therapies,+past+and+present.">Small animal models of heart failure: development of novel therapies, past and present.</a> Circ Heart Fail. 2, 138-144 (2009).</li><li>Gu, W. L., Chen, C. X., Huang, X. Y., Gao, J. 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+effect+of+angoroside+C+on+pressure+overload-induced+ventricular+remodeling+in+rats.">The effect of angoroside C on pressure overload-induced ventricular remodeling in rats.</a> Phytomedicine. 22, 705-712 (2015).</li><li>Zhang, Y., 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=Alteration+of+cardiac+ACE2/Mas+expression+and+cardiac+remodelling+in+rats+with+aortic+constriction.">Alteration of cardiac ACE2/Mas expression and cardiac remodelling in rats with aortic constriction.</a> Chin J Physiol. 57, 335-342 (2014).</li><li>Tardif, K., 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=Nestin+upregulation+characterizes+vascular+remodeling+secondary+to+hypertension+in+the+rat.">Nestin upregulation characterizes vascular remodeling secondary to hypertension in the rat.</a> Am J Physiol Heart Circ Physiol. 308, H1265-H1274 (2015).</li><li>Li, 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=Myeloid+mineralocorticoid+receptor+deficiency+inhibits+aortic+constriction-induced+cardiac+hypertrophy+in+mice.">Myeloid mineralocorticoid receptor deficiency inhibits aortic constriction-induced cardiac hypertrophy in mice.</a> PLoS One. 9, e110950 (2014).</li><li>Ku, H. C., Su, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=DPP4+deficiency+preserved+cardiac+function+in+abdominal+aortic+banding+rats.">DPP4 deficiency preserved cardiac function in abdominal aortic banding rats.</a> PLoS One. 9, e85634 (2014).</li><li>Lee, S. Y., 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=Caffeic+acid+ethanolamide+prevents+cardiac+dysfunction+through+sirtuin+dependent+cardiac+bioenergetics+preservation.">Caffeic acid ethanolamide prevents cardiac dysfunction through sirtuin dependent cardiac bioenergetics preservation.</a> J Biomed Sci. 22, 80 (2015).</li><li>Gs, A. K., Raj, B., Santhosh, K. S., Sanjay, G., Kartha, C. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ascending+aortic+constriction+in+rats+for+creation+of+pressure+overload+cardiac+hypertrophy+model.">Ascending aortic constriction in rats for creation of pressure overload cardiac hypertrophy model.</a> J Vis Exp. , e50983 (2014).</li><li>deAlmeida, A. C., van Oort, R. J., Wehrens, X. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transverse+aortic+constriction+in+mice.">Transverse aortic constriction in mice.</a> J Vis Exp. , e1729 (2010).</li><li>Schaefer, 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=A+New+Animal+Model+for+Investigation+of+Mechanical+Unloading+in+Hypertrophic+and+Failing+Hearts:+Combination+of+Transverse+Aortic+Constriction+and+Heterotopic+Heart+Transplantation.">A New Animal Model for Investigation of Mechanical Unloading in Hypertrophic and Failing Hearts: Combination of Transverse Aortic Constriction and Heterotopic Heart Transplantation.</a> PLoS One. 11, e0148259 (2016).</li><li>Rodriguez-Iturbe, B., Quiroz, Y., Kim, C. H., Vaziri, N. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypertension+induced+by+aortic+coarctation+above+the+renal+arteries+is+associated+with+immune+cell+infiltration+of+the+kidneys.">Hypertension induced by aortic coarctation above the renal arteries is associated with immune cell infiltration of the kidneys.</a> Am J Hypertens. 18, 1449-1456 (2005).</li><li>Ku, H. C., Lee, S. Y., Yang, K. C., Kuo, Y. H., Su, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modification+of+Caffeic+Acid+with+Pyrrolidine+Enhances+Antioxidant+Ability+by+Activating+AKT/HO-1+Pathway+in+Heart.">Modification of Caffeic Acid with Pyrrolidine Enhances Antioxidant Ability by Activating AKT/HO-1 Pathway in Heart.</a> PLoS One. 11, e0148545 (2016).</li><li>Bovill, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intravenous+anesthesia+for+the+patient+with+left+ventricular+dysfunction.">Intravenous anesthesia for the patient with left ventricular dysfunction.</a> Semin Cardiothorac Vasc Anesth. 10, 43-48 (2006).</li><li>Inoko, M., Kihara, Y., Morii, I., Fujiwara, H., Sasayama, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transition+from+compensatory+hypertrophy+to+dilated,+failing+left+ventricles+in+Dahl+salt-sensitive+rats.">Transition from compensatory hypertrophy to dilated, failing left ventricles in Dahl salt-sensitive rats.</a> Am J Physiol. 267, H2471-H2482 (1994).</li><li>Heyen, J. R., 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=Structural,+functional,+and+molecular+characterization+of+the+SHHF+model+of+heart+failure.">Structural, functional, and molecular characterization of the SHHF model of heart failure.</a> Am J Physiol Heart Circ Physiol. 283, H1775-H1784 (2002).</li></ol>

The authors have nothing to disclose.

Hypertensive heart disease, a major health problem that contributes greatly to morbidity and mortality, can lead to cardiac hypertrophy and heart failure5. The pathogenesis and progression of hypertensive heart disease in humans is complex, so an appropriate animal model is critical to investigate the underlying mechanisms and to test novel therapeutics that aim to improve cardiac structure and function5. The abdominal aortic constriction model, which simulates chronic heart disease, is an effective...

心不全は、複雑な臨床症候群、疲労、呼吸困難、運動不耐性を含むそれらの症状、および末梢組織における体液貯留です。これは、先進国1における死亡の主な原因です。別にサルコメアタンパク質またはイオンチャネル2における突然変異によって引き起こされる継承心筋から、心筋機能障害は、高血圧、心臓弁膜症、肥満、および糖尿病3を含む医学的状態、種々の方法により生じさせることができます。最終的には心不全3,4につながる循環要求を満たすために不十分な心臓のポンプ能力、に心筋の構造、電気伝導、およびエネルギー代謝のリードの変化。心不全のメカニズムを調査し、したがって、心臓血管研究の分野において重要です。心不全の進行につながる分子メカニズムを識別することは、最終的に新規治療標的または有用なバイオマーカーの発見を助けることができます<s&gt; 1アップ。ヒト5に心不全とキー臨床的特徴を共有する心不全の動物モデルを開発することが重要です。 心臓肥大及びリモデリングが心不全の発症において重要な役割を果たしています。高血圧性心疾患は、心臓肥大およびヒト患者1に見られる不適応リモデリングの主要な寄与因子です。これらの人間の条件を模倣するために、動物モデルは、多くの場合、外科的処置を介して確立されます。具体的には、横方向または腹部大動脈は、最終的に心臓内の圧負荷につながる左心室に対する抵抗力を高めるために狭窄することができます。この現象は通常、心臓血管系の機能的な要求を満たすために、心臓肥大、心筋細胞の生理学的補償をもたらします。しかし、機能的な需要は、心臓線維症およびcontracにつながる、通常の生理学的代償機構を無効にしますタイルの減損。横方向の大動脈狭窄（TAC）手術は、多くの場合、大動脈弓から開胸、機械換気、および胸腺の分離および脂肪組織を含む複雑な手順を伴います。これとは対照的に、腹部大動脈狭窄は、単純な実験手法6-8を必要とします 。腹部大動脈は、左右の腎動脈との間に、手術中の狭窄です。心肥大およびリモデリングは、数週間腹部大動脈狭窄手術6-8後に観察することができます。彼らは、横大動脈狭窄手術9,10によって生成されたものと同様の堅固な高血圧性心疾患を生じさせます。ここで、我々は、効率的な再現性の高い、かつ低侵襲性の方法を用いて、ラットで腹部大動脈狭窄を行うためのプロトコルを記述します。腎動脈に隣接し、腹部大動脈を4-0絹糸により形成さ0.72ミリメートルループによって絞られます。手術、心臓肥大およびremodelin後10週間gを観察することができます。腹部大動脈狭窄誘発性心肥大のラットモデルは、病気のメカニズムや病態生理学、ならびに潜在的な治療薬の開発を研究するためのプラットフォームを提供します。

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			<td style="width:139px;">Sigma Aldrich&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
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			<td height="21" style="height:21px;width:223px;">Picrosirius red solution&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:139px;">Abcam&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:119px;">ab150681</td>
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			<td height="21" style="height:21px;width:223px;">Cardiac troponin kit&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:139px;">&#160;Abcam&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:119px;">ab200016</td>
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			<td height="42" style="height:42px;width:223px;">Imagequant</td>
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			<td height="21" style="height:21px;width:223px;">Langendorff&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:139px;">&#160;ADInstruments&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:119px;">ML870B2</td>
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a model of cardiac remodeling through constriction of the abdominal aorta in rats

Heart failure is one of the leading causes of death worldwide. It is a complex clinical syndromethat includes fatigue, dyspnea, exercise intolerance, and fluid retention. Changes in myocardial structure, electrical conduction, and energy metabolism develop with heart failure, leading to contractile dysfunction, increased risk of arrhythmias, and sudden death. Hypertensive heart disease is one of the key contributing factors of cardiac remodeling associated with heart failure. The most commonly-used animal model mimicking hypertensive heart disease is created via surgical interventions, such as by narrowing the aorta. Abdominal aortic constriction is a useful experimental technique to induce a pressure overload, which leads to heart failure. The surgery can be easily performed, without the need for chest opening or mechanical ventilation. Abdominal aortic constriction-induced cardiac pathology progresses gradually, making this model relevant to clinical hypertensive heart failure. Cardiac injury and remodeling can be observed 10 weeks after the surgery. The method described here provides a simple and effective approach to produce a hypertensive heart disease animal model that is suitable for studying disease mechanisms and for testing novel therapeutics.

10週間の腹部大動脈狭窄、手術後、得られた心臓の病状を分析しました。心臓の組織学は、体重に対する心臓重量の比を計算することによって、心臓におけるコラーゲンの量を検出することによって測定しました。心臓の損傷は、プラズマ心筋トロポニンの濃度を測定することにより確認しました。 図1Aに示...

Watch this Scientific Journal Video about A Model of Cardiac Remodeling Through Constriction of the Abdominal Aorta in Rats at JoVE.com

A Model of Cardiac Remodeling Through Constriction of the Abdominal Aorta in Rats

A rat model of abdominal aortic constriction that induces cardiac hypertrophy and remodeling is described. An efficient, highly-reproducible, and minimally-invasive method is used to provide a simple yet useful platform for research in myocardial hypertrophy and dysfunction.

ラットの腹部大動脈の収縮を通じて心臓リモデリングのモデル

Heart failure is one of the leading causes of death worldwide. It is a complex clinical syndromethat includes fatigue, dyspnea, exercise intolerance, and ...

a-model-cardiac-remodeling-through-constriction-abdominal-aorta

Research

JoVE Journal

Medicine

9.9K ビュー.  National Taiwan University. A rat model of abdominal aortic constriction that induces cardiac hypertrophy and remodeling is described. An efficient, highly-reproducible, and minimally-invasive method is used to provide a simple yet useful platform for research in myocardial hypertrophy and dysfunction.

ビデオ: A Model of Cardiac Remodeling Through Constriction of the Abdominal Aorta in Rats

Chronic Constriction of the Sciatic Nerve and Pain Hypersensitivity Testing in Rats

Chronic neuropathic pain, resulting from damage to the central or peripheral nervous system, is a prevalent and debilitating condition, affecting 7-18% of the population1,2. Symptoms include spontaneous (tingling, burning, electric-shock like) pain, dysaesthesia, paraesthesia, allodynia (pain resulting from normally non-painful stimuli) and hyperalgesia (an increased response to painful stimuli). The sensory symptoms are co-morbid with behavioural disabilities, such as insomnia and depression. To study chronic neuropathic pain several animal models mimicking peripheral nerve injury have been developed, one of the most widely used is Bennett and Xie's (1988) unilateral sciatic nerve chronic constriction injury (CCI)3 (Figure 1). Here we present a method for performing CCI and testing pain hypersensitivity.
CCI is performed under anaesthesia, with the sciatic nerve on one side exposed by making a skin incision, and cutting through the connective tissue between the gluteus superficialis and biceps femoris muscles. Four chromic gut ligatures are tied loosely around the sciatic nerve at 1 mm intervals, to just occlude but not arrest epineural blood flow. The wound is closed with sutures in the muscle and staples in the skin. The animal is then allowed to recover from surgery for 24 hrs before pain hypersensitivity testing begins.
For behavioural testing, rats are placed into the testing apparatus and are allowed to habituate to the testing procedure. The area tested is the mid-plantar surface of the hindpaw (Figure 2), which falls within the sciatic nerve distribution. Mechanical withdrawal threshold is assessed by mechanically stimulating both injured and uninjured hindpaws using an electronic dynamic plantar von Frey aesthesiometer or manual von Frey hairs4. The mechanical withdrawal threshold is the maximum pressure exerted (in grams) that triggers paw withdrawal. For measurement of thermal withdrawal latency, first described by Hargreaves et al (1988), the hindpaw is exposed to a beam of radiant heat through a transparent glass surface using a plantar analgesia meter5,6. The withdrawal latency to the heat stimulus is recorded as the time for paw withdrawal in both injured and uninjured hindpaws. Following CCI, mechanical withdrawal threshold, as well as thermal withdrawal latency in the injured paw are both significantly reduced, compared to baseline measurements and the uninjured paw (Figure 3). The CCI model of peripheral nerve injury combined with pain hypersensitivity testing provides a model system to investigate the effectiveness of potential therapeutic agents to modify chronic neuropathic pain. In our laboratory, we utilise CCI alongside thermal and mechanical sensitivity of the hindpaws to investigate the role of neuro-immune interactions in the pathogenesis and treatment of neuropathic pain.

Due to the simplicity of surgery and the robust behavioural outcome, chronic constriction of the sciatic nerve is one of the pre-eminent animal models of neuropathic pain. Within 24 hrs following surgery, pain hypersensitivity is established and can be quantitatively measured using a von Frey aesthesiometer (mechanical test) and plantar analgesia meter (thermal test).

ラットの坐骨神経と疼痛過敏テストの慢性的なくびれ

Chronic neuropathic pain, resulting from damage to the central or peripheral nervous system, is a prevalent and debilitating condition, affecting 7-18% of ...

医学

Orthotopic Liver Transplantation in Rats

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat liver transplantation in 1979 by Kamada et al.3, this model has remained the gold standard despite various proposed alternative techniques4. Nevertheless, its broader use is limited by its steep learning curve5.
In this video paper, we show a simple and easy-to-establish revision of Kamada's two-cuff technique. The suprahepatic vena cava anastomosis is performed manually with a running suture, and the vena porta and infrahepatic vena cava anastomoses are performed utilizing a quick-linker cuff system6. Manufacturing the quick-linker kit is shown in a separate video paper.

We present an easy-to-establish revision of the classical two-cuff technique for orthotopic liver transplantation in rat.

ラット同所性肝移植

Clinical progress in the field of liver transplantation has been largely supported by animal models1,2. Since the publication of the first orthotopic rat ...

Ascending Aortic Constriction in Rats for Creation of Pressure Overload Cardiac Hypertrophy Model

Ascending aortic constriction is the most common and successful surgical model for creating pressure overload induced cardiac hypertrophy and heart failure. Here, we describe a detailed surgical procedure for creating pressure overload and cardiac hypertrophy in rats by constriction of the ascending aorta using a small metallic clip. After anesthesia, the trachea is intubated by inserting a cannula through a half way incision made between two cartilage rings of trachea. Then a skin incision is made at the level of the second intercostal space on the left chest wall and muscle layers are cleared to locate the ascending portion of aorta. The ascending aorta is constricted to 50&#8211;60% of its original diameter by application of a small sized titanium clip. Following aortic constriction, the second and third ribs are approximated with prolene sutures. The tracheal cannula is removed once spontaneous breathing was re-established. The animal is allowed to recover on the heating pad by gradually lowering anesthesia. The intensity of pressure overload created by constriction of the ascending aorta is determined by recording the pressure gradient using trans-thoracic two dimensional Doppler-echocardiography. Overall this protocol is useful to study the remodeling events and contractile properties of the heart during the gradual onset and progression from compensated cardiac hypertrophy to heart failure stage.

We describe a stepwise procedure for creating pressure overload and left ventricular hypertrophy in Wistar rats by constriction of the ascending aorta using a small metallic clip. This model is extensively used for studying remodeling changes during cardiac hypertrophy and for identifying and evaluating strategies for regression of such changes.

圧負荷心肥大モデルの作成のためのラットの大動脈狭窄昇順

Ascending aortic constriction is the most common and successful surgical model for creating pressure overload induced cardiac hypertrophy and heart ...

Assessment of Right Ventricular Structure and Function in Mouse Model of Pulmonary Artery Constriction by Transthoracic Echocardiography

Emerging clinical data support the notion that RV dysfunction is critical to the pathogenesis of cardiovascular disease and heart failure1-3. Moreover, the RV is significantly affected in pulmonary diseases such as pulmonary artery hypertension (PAH). In addition, the RV is remarkably sensitive to cardiac pathologies, including left ventricular (LV) dysfunction, valvular disease or RV infarction4. To understand the role of RV in the pathogenesis of cardiac diseases, a reliable and noninvasive method to access the RV structurally and functionally is essential.
A noninvasive trans-thoracic echocardiography (TTE) based methodology was established and validated for monitoring dynamic changes in RV structure and function in adult mice. To impose RV stress, we employed a surgical model of pulmonary artery constriction (PAC) and measured the RV response over a 7-day period using a high-frequency ultrasound microimaging system. Sham operated mice were used as controls. Images were acquired in lightly anesthetized mice at baseline (before surgery), day 0 (immediately post-surgery), day 3, and day 7 (post-surgery). Data was analyzed offline using software.
Several acoustic windows (B, M, and Color Doppler modes), which can be consistently obtained in mice, allowed for reliable and reproducible measurement of RV structure (including RV wall thickness, end-diastolic&#160;and end-systolic dimensions), and function (fractional area change, fractional shortening, PA peak velocity, and peak pressure gradient) in normal mice and following PAC.
Using this method, the pressure-gradient resulting from PAC was accurately measured in real-time using Color Doppler mode and was comparable to direct pressure measurements performed with a Millar high-fidelity microtip catheter. Taken together, these data demonstrate that RV measurements obtained from various complimentary views using echocardiography are reliable, reproducible and can provide insights regarding RV structure and function. This method will enable a better understanding of the role of RV cardiac dysfunction.

Right ventricle (RV) dysfunction is critical to the pathogenesis of cardiovascular disease, yet limited methodologies are available for its evaluation. Recent advances in ultrasound imaging provide a noninvasive and accurate option for longitudinal RV study. Herein, we detail a step-by-step echocardiographic method using a murine model of RV pressure overload.

経胸壁心エコー検査による肺動脈狭窄のマウスモデルにおける右室の構造と機能の評価

Emerging clinical data support the notion that RV dysfunction is critical to the pathogenesis of cardiovascular disease and heart failure1-3. Moreover, ...

Visualization of Streptococcus pneumoniae within Cardiac Microlesions and Subsequent Cardiac Remodeling

During bacteremia Streptococcus pneumoniae can translocate across the vascular endothelium into the myocardium and form discrete bacteria-filled microscopic lesions (microlesions) that are remarkable due to the absence of infiltrating immune cells. Due to their release of cardiotoxic products, S. pneumoniae within microlesions are thought to contribute to the heart failure that is frequently observed during fulminate invasive pneumococcal disease in adults. Herein is demonstrated a protocol for experimental mouse infection that leads to reproducible cardiac microlesion formation within 30 hr. Instruction is provided on microlesion identification in hematoxylin &#38; eosin stained heart sections and the morphological distinctions between early and late microlesions are highlighted. Instruction is provided on a protocol for verification of S. pneumoniae within microlesions using antibodies against pneumococcal capsular polysaccharide and immunofluorescent microscopy. Last, a protocol for antibiotic intervention that rescues infected mice and for the detection and assessment of scar formation in the hearts of convalescent mice is provided. Together, these protocols will facilitate the investigation of the molecular mechanisms underlying pneumococcal cardiac invasion, cardiomyocyte death, cardiac remodeling as a result of exposure to S. pneumoniae, and the immune response to the pneumococci in the heart.

Visualization of Streptococcus pneumoniae within Cardiac Microlesions and Subsequent Cardiac Remodeling

Streptococcus pneumoniae forms discrete non-purulent microscopic lesions in the heart. Outlined is the protocol for a murine model of cardiac microlesion formation. Instruction is provided on microlesion visualization using microscopy, discrimination between early and late microlesions, and methods to detect cardiac remodeling in hearts of convalescent animals.

の可視化<em&gt;肺炎球菌</em&gt;心臓Microlesionsとその後の心臓リモデリングの中

During bacteremia Streptococcus pneumoniae can translocate across the vascular endothelium into the myocardium and form discrete bacteria-filled ...

Assessment of Cardiac Morphological and Functional Changes in Mouse Model of Transverse Aortic Constriction by Echocardiographic Imaging

Transverse aortic constriction (TAC) in mice has been used as a valuable model to study mechanisms of cardiac hypertrophy and heart failure1. A reliable noninvasive method is essential to assess real-time cardiac morphological and functional changes in animal models of heart disease. Transthoracic echocardiography represents an important tool for noninvasive assessment of cardiac structure and function2. Here we used a high-resolution ultrasound imaging system to monitor myocardial remodeling and heart failure progression over time in a mouse model of TAC. B-mode, M-mode, and Doppler imaging were used to precisely assess cardiac hypertrophy, ventricular dilatation, and functional deterioration in mice following TAC. Color and pulse wave (PW) Doppler imaging was used to noninvasively measure pressure gradient across the aortic constriction created by TAC and to assess transmitral blood flow in mice. Thus transthoracic echocardiographic imaging provides comprehensive noninvasive measurements of cardiac dimensions and function in mouse models of heart disease.

The goal of this protocol is to noninvasively assess cardiac structural and functional changes in a mouse model of heart disease created by transverse aortic constriction, using B- and M-mode echocardiography and color/pulse wave Doppler imaging.

心臓形態学的および心エコーイメージングによって横大動脈狭窄のマウスモデルにおける機能の変更の評価

Transverse aortic constriction (TAC) in mice has been used as a valuable model to study mechanisms of cardiac hypertrophy and heart failure1. A reliable ...

Balloon-based Injury to Induce Myointimal Hyperplasia in the Mouse Abdominal Aorta

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine balloon denudation model, which is comparable with established vessel injury models in large animals. The aorta denudation model with balloon catheters mimics the clinical setting and leads to comparable pathobiological and physiological changes. Briefly, after performing a horizontal incision in the aorta abdominalis, a balloon catheter will be inserted into the vessel, inflated, and introduced retrogradely. Inflation of the balloon will lead to intima injury and overdistension of the vessel. After removing the catheter, the aortic incision will be closed with single stiches. The model shown in this article is reproducible, easy to perform, and can be established quickly and reliably. It is especially suitable for evaluating expensive experimental therapeutic agents, which can be applied in an economical fashion. By using different knockout-mouse strains, the impact of different genes on MH development can be assessed.

This article demonstrates a murine model to study the development of myointimal hyperplasia (MH) after aortic balloon injury.

マウスの腹部大動脈 Myointimal 過形成を誘導するために気球を利用した傷害

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine ...

A Closed-chest Model to Induce Transverse Aortic Constriction in Mice

Research on cardiac hypertrophy and heart failure is frequently based on pressure overload mouse models induced by TAC. The standard procedure is to perform a partial thoracotomy to visualize the transverse aortic arch. However, the surgical trauma caused by the thoracotomy in open-chest models changes the respiratory physiology as the ribs are dissected and left unattached after chest closure. To prevent this, we established a minimally invasive, closed chest approach via lateral thoracotomy. Herein we approach the aortic arch via the 2nd intercostal space without entering the chest cavities, leaving the mouse with a less traumatic injury to recover from. We perform this operation using standard laboratory settings for open chest TAC procedures with equal survival rates. Apart from maintaining physiological breathing patterns due to the closed chest approach, the mice seem to benefit by showing rapid recovery, as the less invasive technique appears to facilitate a fast healing process and to reduce immune response after trauma.

Here, we present a protocol of Transverse Aortic constriction (TAC) via a lateral thoracotomy. This technique is a minimally invasive, closed chest surgical procedure aiming to simulate pressure overload and heart failure in mice utilizing standard TAC laboratory settings.

マウスで横方向の大動脈狭窄を誘導するために閉じて胸モデル

Research on cardiac hypertrophy and heart failure is frequently based on pressure overload mouse models induced by TAC. The standard procedure is to ...

Ultrasound Imaging of the Thoracic and Abdominal Aorta in Mice to Determine Aneurysm Dimensions

Contemporary high-resolution ultrasound instruments have sufficient resolution to facilitate the measurement of mouse aortas. These instruments have been widely used to measure aortic dimensions in mouse models of aortic aneurysms. Aortic aneurysms are defined as permanent dilations of the aorta, which occur most frequently in the ascending and abdominal regions. Sequential measurements of aortic dimensions by ultrasound are the principal approach for assessing the development and progression of aortic aneurysms in vivo. Although many reported studies used ultrasound imaging to measure aortic diameters as a primary endpoint, there are confounding factors, such as probe position and cardiac cycle, that may impact the accuracy of data acquisition, analysis, and interpretation. The purpose of this protocol is to provide a practical guide on the use of ultrasound to measure the aortic diameter in a reliable and reproducible manner. This protocol introduces the preparation of mice and instruments, the acquisition of appropriate ultrasound images, and data analysis.

Ultrasound imaging has become a common modality to determine the luminal dimensions of thoracic and abdominal aortic aneurysms in mice. This protocol describes the procedure to acquire reliable and reproducible two-dimensional ultrasound images of the ascending and abdominal aorta in mice.

マウス脳動脈瘤のサイズを調べるために胸部、腹部の大動脈の超音波断層像

Contemporary high-resolution ultrasound instruments have sufficient resolution to facilitate the measurement of mouse aortas. These instruments have been ...

Implantation of Human-Sized Coronary Stents into Rat Abdominal Aorta Using a Trans-Femoral Access

Percutaneous coronary intervention (PCI), combined with the deployment of a coronary stent, represents the gold standard in interventional treatment of coronary artery disease. In-stent restenosis (ISR) is determined by an excessive proliferation of neointimal tissue within the stent and limits the long-term success of stents. A variety of animal models have been used to elucidate pathophysiological processes underlying in-stent restenosis (ISR), with the porcine coronary and the rabbit iliac artery models being the most frequently used. Murine models provide the advantages of high throughput, ease of handling and housing, reproducibility, and a broad availability of molecular markers. The apolipoprotein E deficient (apoE-/- ) mouse model has been widely used to study cardiovascular diseases. However, stents must be miniaturized to be implanted into mice, involving important changes of their mechanical and (potentially) biological properties. The use of apoE-/- rats can overcome these shortcomings as apoE-/- rats allow for the evaluation of human-sized coronary stents while at the same time providing an atherogenic phenotype. This makes them an excellent and reliable model to investigate ISR after stent implantation. Here, we describe, in detail, the implantation of commercially available human coronary stents into the abdominal aorta of rats with an apoE-/- background using a trans-femoral access.

This protocol describes the implantation of human coronary stents into the abdominal aorta of rats with an apoE-/- background using a trans-femoral access. Compared with other animal models, murine models carry the advantages of high throughput, reproducibility, ease of handling and housing, and a broad availability of molecular markers.

トランス・大腿部アクセスを用いたヒトサイズの冠動脈ステントのラット腹部大動脈への移植

Percutaneous coronary intervention (PCI), combined with the deployment of a coronary stent, represents the gold standard in interventional treatment of ...