The work was supported by the intramural research program of the NIH, National Institute on Aging, and by a Priority Research Centers Program grant from the National Research Foundation (NRF-2009-0093812) funded by the Ministry of Science, Information and Communication Technology, &amp; Future Planning, the Republic of Korea (H.T.). We thank Dr. Han-sol Park for putting "material" part together for the manuscript.

注：ラットバルーン損傷モデルおよび組換えのsRAGEと超音波超音波検査の研究の注入を含む、関連する手順の使用は、国立老化研究所の動物実験委員会（ACUC）、NIHによって承認されています。 1.手術前の準備<ol><li>インストゥルメント、外科プラットフォーム、および個人装備。 <ol><li>この手順で使用するすべての手術器具および試薬のための材料および機器の表を参照してください。外科手術の前に、すべての手術器具をオートクレーブ。 注：同じ日に実行される複数の手術の場合、手術の間に器具を滅菌するために予備加熱したガラスビーズ滅菌器を使用しています。 </li><li> 70％アルコールでオペレーティングプラットフォームの表面を殺菌します。手術用ガウン、ヘアカバー、外科手術用マスク、保護眼鏡、と手袋を着用してください。追加の私たちのための外科的なプ​​ラットフォームの近くに滅菌手袋のボックスを残します電子。 </li></ol></li><li>バルーンカテーテルの準備。 <ol><li>滅菌水で1ミリリットル注射器を記入し、滅菌水で満たされた2方コックに注射器を取り付けます。静かにバルーンカテーテルのルアーロック部を記入し、カテーテルの開口部から気泡を除去するためにストップコックを通して水をプッシュ。 </li><li>バルーンを保証するテストバルーン膨張は、水0.02mlのことで膨張させることができます。乾燥や汚染を避けるために手術前に滅菌水の容器にバルーンの先端に保管してください。 </li></ol></li><li>麻酔と動物の準備。 <ol><li> Wistar系雄性ラット（400〜450グラム）を計量し、IDを確認するために、耳のタグを確認してください。このようなつま先や尾をつまんとしてペダル引っ込め反射をチェックすることによって、鎮静の深さを確認し、誘導チャンバー内にラットを置き、その後、気化器をオンにして1.5〜2.0 Lに3.5から4.0パーセントにイソフルランレベルと酸素レベルを調整動物の。 </li><li>麻酔に達した後（ <em&gt;動物）はペダル引っ込め反射を欠いている。すなわち、外科的なプラットフォームに動物を転送し、麻酔気化器に接続するノーズコーンとラットをフック。 L.は、定期的にラットが完全にリラックスして不動であることを確実にするために、動物のつま先や尾をつまんで鎮静の深さを確認して0.6から1.0に1.5から2.0と酸素にイソフルラン濃度を調整します。 </li><li>麻酔の外科的なレベルに到達した後、単に動物のバリカンと胸骨の上までchinから腹側ネック領域を剃るし、ポビドンヨードおよび70％エタノールで3回の1:10希釈液を交互に剃毛した皮膚を綿棒。 </li><li>乾燥からそれらを防ぐために、綿棒で動物の開いた目に点眼液を置きます。動作中の乾燥を避けるために、水を飲んで動物の舌と口を湿ら。 </li><li>無菌手術用シートでラットを覆い、ネック領域を露出させるためにシートをカット。仰臥位のOに動物を置きます外科医に向かって頭を手術用プラットフォーム上のパッド加熱ナ。テープを使用して、手術用プラットフォームに動物の手足を修正しました。 </li></ol></li></ol> 2.外科的処置<ol><li>左頸動脈の解剖と単離<ol><li>ちょうど胸郭の上に尾部に向かう方向に胸骨の先頭にすべての方法を顎の下のストレート切開を作るためにメスを使用してください。ぶっきらぼうに筋層を露出させるために皮膚の下唾液腺組織を分析する7S鉗子を使用してください。頸動脈管構造とネック領域における迷走神経へのアクセスを獲得するために別の筋肉組織。 </li><li>ぶっきらぼうに頸動脈を周囲の組織を解剖し、慎重に任意の損傷を避けるために迷走神経と血管筋膜を分離します。継続的にダウンして胸骨の内部および外部の支店および総頸動脈を露出させるために頸動脈分岐部まで解剖します。 </li></ol></li><li> Arteriotの調製大丸有<ol><li>後退や止血のために、左総頸動脈に最も近位のサイトで合字を置き、分岐点に位置即時遠位にある別のリガチャー。 </li><li>頭に向かって動脈を後退させるためにできる限りの分岐部から外頸動脈で4.0絹縫合糸を配置します。永久昇順咽頭、後頭動脈漏れを回避するための優れた甲状腺を含む外頸動脈に沿って4.0絹縫合糸を用いて、小動脈を結紮。 </li><li>重大な逆行性の血液の損失を回避するために、内頸動脈の周りに4.0絹縫合糸を配置します。通常、内部分岐の利用可能な長さは最小です。内頚動脈を可視化するために、静かに右に、上にある頸動脈を後退させることにより可能な限り内頸動脈を露出させます。 </li></ol></li><li>バルーンカテーテルの導入<ol><li>総頸動Aの近位縫合糸を撤回rteryとは、一時的に血流を止めるために、血管に動脈クランプを配置します。できるだけ縫合糸のよう遠位位置で外頸動脈に（1/3動脈の円周の1/4）に小切開を加えます。 </li><li>優しく切開部に未膨張バルーンカテーテルを挿入し、総頸動脈に動脈クランプに近い内腔にカテーテルを進めます。クランプを外し、さらに切開から約35〜40ミリメートル、大動脈弓にバルーンカテーテルを進めます。 </li></ol></li><li>バルーン傷害<ol><li>手動で添付シリンジを介して0.02ミリリットルの容量にカテーテルを膨張し、その後、注射器とカテーテルとの間のコックをロックします。ゆっくりと回転してカテーテルを引き抜きます。カテーテルはarteriotomic切開部に近接している場合には、バルーンを収縮させます。元の場所にカテーテルを進めます。 </li><li>合計3回2.4.1を繰り返します。慎重に動脈内腔からカテーテルを除去します。 </li></ol></li><li>手術創をクローズアップ<ol><li>縫合糸で動脈切開を閉じます。血流を回復するために内頸動脈をリリース。動脈出血やリークを確認してください。出血が発生した場合は、出血を止めるために、ビット圧力でガーゼを適用します。 </li><li>手続きのために使用される縫合糸およびクランプの残りの部分を削除します。 6.0縫合糸で腺組織を閉じます。皮膚の縫合糸または創傷クリップで皮膚を閉じます。イソフルランの電源を切り、数分間、純酸素で動物を残します。 </li></ol></li></ol> 3.治療薬の投与は、鎮痛薬、手術後のケア、および安楽死<ol><li>治療薬や鎮痛剤の投与。 <ol><li>手術後、筋肉内にブプレノルフィン（0.03ミリグラム/キログラム）を注入します。手術中に血液の損失を補償するために、5ミリリットル予め温めた滅菌生理食塩水を皮下に注入します。腹膜へのsRAGE（0.5 ngの/ g）を（治療薬）を注入します。 </li><li> OPHで再び目を湿らthalmic軟膏。熱パッドの上にラットを置き、胸骨横臥位を維持するために完全な意識を取り戻すまで、動物を監視します。 </li><li>きれいな寝具を使用してケージにラットを戻し、バックビバリウムにケージを持参。出席獣医師や世話人を警告するためにケージに特殊な観測カードを置きます。 </li></ol></li><li>術後のクリーンアップ<ol><li>ゴミや特殊な廃棄容器へのいずれかの針らパッド、綿棒、を含む使用済みの外科材料を入れてください。手術プラットフォームを消毒し、手術器具を清掃してください。 </li></ol></li><li>術後のケア<ol><li> 48時間後に手術期間中にブプレノルフィン（0.03ミリグラム/キログラム）一日二回（BID）を管理します。このような不動、養うことができないこと、hutched姿勢、フリル毛皮、としかめっ面のような苦痛の一般的な徴候のための48時間術後期間中に密接に動物を監視します。創傷赤みのための条件、腫れ、裂開、および感染症を監視します。 注意：追加の治療は、獣医師の勧告に出席に応じて処方されることがあります。 </li><li>獣医師の勧告に出席に応じて縫合または創傷クリップを削除します。 </li></ol></li><li>安楽死<ol><li>手術の2週間後に、22を分析し 、死後ヒスト形態学のための頸動脈を分離するために、ラットを安楽死させます。呼吸の完全な停止するまで、少なくとも2分間、空気を飽和5から20までパーセントイソフルランを含むチャンバーにラットを置きます。前の器官および組織の切開に死を確認します。 </li></ol></li></ol>

<ol>
	<li>Chaabane, C., Otsuka, F., Virmani, R., Bochaton-Piallat, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biological+responses+in+stented+arteries.">Biological responses in stented arteries.</a> Cardiovasc Res. 99, 353-363 (2013).</li><li>Goel, S. A., Guo, L. W., Liu, B., Kent, K. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanisms+of+post-intervention+arterial+remodelling.">Mechanisms of post-intervention arterial remodelling.</a> Cardiovasc Res. 96, 363-371 (2012).</li><li>Khan, R., Agrotis, A., Bobik, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Understanding+the+role+of+transforming+growth+factor-beta1+in+intimal+thickening+after+vascular+injury.">Understanding the role of transforming growth factor-beta1 in intimal thickening after vascular injury.</a> Cardiovasc Res. 74, 223-234 (2007).</li><li>Schillinger, M., Minar, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Restenosis+after+percutaneous+angioplasty:+the+role+of+vascular+inflammation.">Restenosis after percutaneous angioplasty: the role of vascular inflammation.</a> Vasc Health Risk Manag. 1, 73-78 (2005).</li><li>Katz, G., Harchandani, B., Shah, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drug-eluting+stents:+the+past,+present,+and+future.">Drug-eluting stents: the past, present, and future.</a> Curr Atheroscler Rep. 17, 485 (2015).</li><li>Jain, M., Singh, A., Singh, V., Barthwal, M. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Involvement+of+Interleukin-1+Receptor-Associated+Kinase-1+in+Vascular+Smooth+Muscle+Cell+Proliferation+and+Neointimal+Formation+After+Rat+Carotid+Injury.">Involvement of Interleukin-1 Receptor-Associated Kinase-1 in Vascular Smooth Muscle Cell Proliferation and Neointimal Formation After Rat Carotid Injury.</a> Arterioscler Thromb Vasc Biol. , (2015).</li><li>Lee, K. 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=Carvacrol+inhibits+atherosclerotic+neointima+formation+by+downregulating+reactive+oxygen+species+production+in+vascular+smooth+muscle+cells.">Carvacrol inhibits atherosclerotic neointima formation by downregulating reactive oxygen species production in vascular smooth muscle cells.</a> Atherosclerosis. 240, 367-373 (2015).</li><li>Li, G., Chen, S. J., Oparil, S., Chen, Y. F., Thompson, 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=Direct+in+vivo+evidence+demonstrating+neointimal+migration+of+adventitial+fibroblasts+after+balloon+injury+of+rat+carotid+arteries.">Direct in vivo evidence demonstrating neointimal migration of adventitial fibroblasts after balloon injury of rat carotid arteries.</a> Circulation. 101 (12), 1362-1365 (2000).</li><li>Noda, 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=New+endoplasmic+reticulum+stress+regulator,+gipie,+regulates+the+survival+of+vascular+smooth+muscle+cells+and+the+neointima+formation+after+vascular+injury.">New endoplasmic reticulum stress regulator, gipie, regulates the survival of vascular smooth muscle cells and the neointima formation after vascular injury.</a> Arterioscler Thromb Vasc Biol. 35 (5), 1246-1253 (2015).</li><li>Deuse, 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=Dichloroacetate+prevents+restenosis+in+preclinical+animal+models+of+vessel+injury.">Dichloroacetate prevents restenosis in preclinical animal models of vessel injury.</a> Nature. 509 (7502), 641-644 (2014).</li><li>Guo, J., 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=p55gamma+functional+mimetic+peptide+N24+blocks+vascular+proliferative+disorders.">p55gamma functional mimetic peptide N24 blocks vascular proliferative disorders.</a> J Mol Med (Berl). , (2015).</li><li>Oh, C. J., 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=Dimethylfumarate+attenuates+restenosis+after+acute+vascular+injury+by+cell-specific+and+Nrf2-dependent+mechanisms.">Dimethylfumarate attenuates restenosis after acute vascular injury by cell-specific and Nrf2-dependent mechanisms.</a> Redox Biol. 2, 855-864 (2014).</li><li>Tae, H. J., 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+N-glycoform+of+sRAGE+is+the+key+determinant+for+its+therapeutic+efficacy+to+attenuate+injury-elicited+arterial+inflammation+and+neointimal+growth.">The N-glycoform of sRAGE is the key determinant for its therapeutic efficacy to attenuate injury-elicited arterial inflammation and neointimal growth.</a> J Mol Med (Berl. 91 (12), 1369-1381 (2013).</li><li>Zhou, 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=Receptor+for+AGE+(RAGE)+mediates+neointimal+formation+in+response+to+arterial+injury.">Receptor for AGE (RAGE) mediates neointimal formation in response to arterial injury.</a> Circulation. 107 (17), 2238-2243 (2003).</li><li>Lindner, V., Fingerle, J., Reidy, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+model+of+arterial+injury.">Mouse model of arterial injury.</a> Circ Res. 73 (5), 792-796 (1993).</li><li>Le, V., Johnson, C. G., Lee, J. D., Baker, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+model+of+femoral+artery+wire+injury+with+implantation+of+a+perivascular+drug+delivery+patch.">Murine model of femoral artery wire injury with implantation of a perivascular drug delivery patch.</a> J Vis Exp. (96), e52403 (2015).</li><li>Iannaccone, P. M., Jacob, 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=Rats!+.">Rats! .</a> Dis Model Mech. 2 (5-6), 206-210 (2009).</li><li>Fu, Y., Sander, J. D., Reyon, D., Cascio, V. M., Joung, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improving+CRISPR-Cas+nuclease+specificity+using+truncated+guide+RNAs.">Improving CRISPR-Cas nuclease specificity using truncated guide RNAs.</a> Nat Biotechnol. 32 (3), 279-284 (2014).</li><li>Sander, J. D., Joung, J. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CRISPR-Cas+systems+for+editing,+regulating+and+targeting+genomes.">CRISPR-Cas systems for editing, regulating and targeting genomes.</a> Nat Biotechnol. 32 (4), 347-355 (2014).</li><li>Tulis, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+carotid+artery+balloon+injury+model.">Rat carotid artery balloon injury model.</a> Methods Mol Med. 139, 1-30 (2007).</li><li>Zhang, W., Trebak, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascular+balloon+injury+and+intraluminal+administration+in+rat+carotid+artery.">Vascular balloon injury and intraluminal administration in rat carotid artery.</a> J Vis Exp. (94), (2014).</li><li>Tae, H. J., 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=Vessel+ultrasound+sonographic+assessment+of+soluble+receptor+for+advanced+glycation+end+products+efficacy+in+a+rat+balloon+injury+model.">Vessel ultrasound sonographic assessment of soluble receptor for advanced glycation end products efficacy in a rat balloon injury model.</a> Curr Ther Res Clin Exp. 76, 110-115 (2014).</li><li>Tae, H. J., 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=Chronic+treatment+with+a+broad-spectrum+metalloproteinase+inhibitor,+doxycycline,+prevents+the+development+of+spontaneous+aortic+lesions+in+a+mouse+model+of+vascular+Ehlers-Danlos+syndrome.">Chronic treatment with a broad-spectrum metalloproteinase inhibitor, doxycycline, prevents the development of spontaneous aortic lesions in a mouse model of vascular Ehlers-Danlos syndrome.</a> J Pharmacol Exp Ther. 343 (1), 246-251 (2012).</li><li>Sakaguchi, 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=Central+role+of+RAGE-dependent+neointimal+expansion+in+arterial+restenosis.">Central role of RAGE-dependent neointimal expansion in arterial restenosis.</a> J Clin Invest. 111 (7), 959-972 (2003).</li></ol>

著者は、彼らが競合する金融利害関係を持たないことを宣言します。

頚動脈の動脈内腔で壁画内皮を除去損傷を生成するためにバルーンを膨張させるために使用される二つの方法がありました。一つは、液体20を取り付けた注射器を充填することであり、他方は空気圧21を使用することです。我々は、正確な液量（0.02 ml）を各手順に使用されるので、液体が充填された注射器を使用することを好みます。これは、処置を受けて各動物における損?...

血管形成術は、アテローム性動脈硬化症のような病理学的状態から得られる狭めたり閉塞した動脈を広げるために使用される血管内処置です。血管形成術の一つの一般的な合併症は、外科的傷害とその後の炎症誘発性血管リモデリングにより発生した後の運用の新生内膜過形成、または再狭窄、です。これらの条件は、血管平滑筋細胞の増殖、および複数の病態生理学的な結果1-3につながります。新生内膜過形成の容器を再厚く、かつ1年以内に60％まで血管形成後の患者の中で起こります。したがって、再狭窄は、広く使用されている血管形成術手順4の主要な後退です。薬剤溶出ステントの植え込みは、再狭窄を防止するのに役立つかもしれないが、唯一の選択された候補は、この費用のかかる手順5を受けることができます。 動物および臨床研究の両方が慢性炎症がvascuによって生成されたことを確立しましたLAR傷害および/ ​​または手術創は、血管形成後の新生内膜増殖2,4のための主要な刺激として機能します。ラット頸動脈バルーン損傷モデルは、臨床的状況を模倣し、従って、血管リモデリングおよび血管細胞増殖6-9に関与する細胞因子を同定するための貴重なモデルシステムとして機能します。このモデル系は、また、前臨床翻訳研究10-14における新生内膜増殖を抑制する薬物および治療 ​​薬のために非常に有用で評価するためのツール及び/又はスクリーンです。 マウス頸動脈ワイヤ傷害モデル15およびマウス大腿動脈ワイヤ傷害モデル16に比べて、ラット頸動脈バルーン傷害モデルが与え損傷の再現性を容易にする外科的処置を容易にするためにサイズが十分に大きいという利点を有します。これはADDITための一次細胞の多く（ 例えば 、血管平滑筋細胞、内皮細胞）を提供することができますional インビトロ血管リモデリングを支配する分子機構を描くための研究。重要なことには、マウスに比べて、ラットにも生理学的および毒性試験17のためのより良いモデルであることが知られています。ラットモデルの欠点または制限が変更された遺伝的および遺伝子ノックアウトモデルの欠如であるが、この欠点は、ラットゲノム配列の可用性と、このようなレンダリングCRISPR-CAS技術などの強力なゲノム編集ツールの最近の開発によって克服することができます異なるモデル系18,19におけるゲノム配列の広大な範囲の可能な操作。 ラットバルーン傷害モデルでは、複数のラボや様々な包括的なプロトコルで使用されてきたが20,21を公開されている、このプロトコルは、手術前の準備でより詳細な情報を提供することを目的とし、この外科的な練習をセットアップするには、この手順に新しい研究者を案内することができます。また、トンの術後のケアを重視します生きた動物13,22だけでなく、動脈リモデリングに対する治療効果の死後の病理学的および組織形態学的分析だけでなく、超音波超音波検査の研究ができます彼の動物。

<table><tbody><tr><td>2 F Fogarty balloon embolectomy catheter&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;</td><td>Edwards Lifesciences&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;</td><td></td><td></td></tr><tr><td>Standard scalpel</td><td>Fine Science Tools</td><td></td><td></td></tr><tr><td>Small&amp;nbsp; curved forceps (Large radius Dumont#7shanks curved)&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;</td><td>Fine Science Tools&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;</td><td></td><td></td></tr><tr><td>Large, medium and small micro-scissors</td><td>Roboz</td><td></td><td></td></tr><tr><td>Needles (20 G)&amp;nbsp;&amp;nbsp;</td><td>TycoHealthcare</td><td></td><td></td></tr><tr><td>Micro-surgery forceps with micro-blunted atraumatic tips</td><td>Fine Science Tools&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;</td><td></td><td></td></tr><tr><td>Atraumatic straight small arterial clamps&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;</td><td>Fine Science Tools&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;</td><td></td><td></td></tr><tr><td>Retractor&amp;nbsp; with maximum spread 5.5 cm long blunt teeth</td><td>Fine Science Tools&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;</td><td></td><td></td></tr><tr><td>Silk suture (4.0 and&amp;nbsp; 6.0 )</td><td>Fine Science Tools&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;</td><td></td><td></td></tr><tr><td>Syringe (1.0 ml)&amp;nbsp;</td><td>BD&amp;nbsp;</td><td></td><td></td></tr><tr><td>Curity gauze sponges</td><td>AllegroMedical</td><td></td><td></td></tr><tr><td>Cotton tip applicators sterile and non-sterile</td><td>Puritan Medical Products</td><td></td><td></td></tr><tr><td>Compact hot bead sterilizer</td><td>Fine Science Tools</td><td></td><td></td></tr><tr><td>Self-regulating heating pad</td><td>Fine Science Tools&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;</td><td></td><td></td></tr><tr><td>ADS200 anesthesia system/ventilator</td><td>Paragon Medical</td><td></td><td></td></tr><tr><td>Isoflurane (forane), liquid form</td><td>Baxter</td><td></td><td></td></tr><tr><td>Sodium chloride 0.9% (Saline)&amp;nbsp;</td><td>Hospira</td><td></td><td></td></tr><tr><td>Buprenex (buprenorphine)&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;&amp;nbsp;</td><td>Reckitt Benckiser Healthcare (UK) Ltd.&amp;nbsp;</td><td></td><td></td></tr><tr><td>70% alcohol</td><td>Fisher</td><td></td><td></td></tr><tr><td>1:10 Betadine</td><td>Fisher</td><td></td><td></td></tr></tbody></table>

a rat carotid balloon injury model to test anti-vascular remodeling therapeutics

The rat carotid balloon injury is a well-established surgical model that has been used to study arterial remodeling and vascular cell proliferation. It is also a valuable model system to test, and to evaluate therapeutics and drugs that negate maladaptive remodeling in the vessel. The injury, or barotrauma, in the vessel lumen caused by an inflated balloon via an inserted catheter induces subsequent neointimal growth, often leading to hyperplasia or thickening of the vessel wall that narrows, or obstructs the lumen. The method described here is sufficiently sensitive, and the results can be obtained in relatively short time (2 weeks after the surgery). The efficacy of the drug or therapeutic against the induced-remodeling can be evaluated either by the post-mortem pathological and histomorphological analysis, or by ultrasound sonography in live animals. In addition, this model system has also been used to determine the therapeutic window or the time course of the administered drug. These studies can leadto the development of a better administrative strategy and a better therapeutic outcome. The procedure described here provides a tool for translational studies that bring drug and therapeutic candidates from bench research to clinical applications.

二週間バルー​​ン損傷後、ラットを安楽死させられ、頸動脈はヒスト形態素解析のために単離されます。両方とも、左頸動脈および非作動右動脈は、断面に処理され、パラフィン包埋されている操作しました。パラフィン次いで、サンプルをさらに薄い切片であり、そしてヘマトキシリン​​ - エオシン（H＆E）で染色しました。ヒスト形態学的分析は、デジタル?...

Watch this Scientific Journal Video about A Rat Carotid Balloon Injury Model to Test Anti-vascular Remodeling Therapeutics at JoVE.com

A Rat Carotid Balloon Injury Model to Test Anti-vascular Remodeling Therapeutics

The rat carotid balloon injury model described below allows researchers to evaluate drugs or therapeutics that negate injury-induced arterial hyperplasia. Detailed pre-surgical preparation, surgical procedure, and post-surgical cares of the animal are described.

ラット頸動脈バルーン損傷モデルは、抗血管リモデリングの治療薬をテストします

The rat carotid balloon injury is a well-established surgical model that has been used to study arterial remodeling and vascular cell proliferation. It is ...

a-rat-carotid-balloon-injury-model-to-test-anti-vascular-remodeling

Research

JoVE Journal

Medicine

8.7K ビュー.  National Institute on Aging. The rat carotid balloon injury model described below allows researchers to evaluate drugs or therapeutics that negate injury-induced arterial hyperplasia. Detailed pre-surgical preparation, surgical procedure, and post-surgical cares of the animal are described. 

ビデオ: A Rat Carotid Balloon Injury Model to Test Anti-vascular Remodeling Therapeutics

A Model of Disturbed Flow-Induced Atherosclerosis in Mouse Carotid Artery by Partial Ligation and a Simple Method of RNA Isolation from Carotid Endothelium

Despite the well-known close association, direct evidence linking disturbed flow to atherogenesis has been lacking. We have recently used a modified version of carotid partial ligation methods [1,2] to show that it acutely induces low and oscillatory flow conditions, two key characteristics of disturbed flow, in the mouse common carotid artery. Using this model, we have provided direct evidence that disturbed flow indeed leads to rapid and robust atherosclerosis development in Apolipoprotein E knockout mouse [3]. We also developed a method of endothelial RNA preparation with high purity from the mouse carotid intima [3]. Using this mouse model and method, we found that partial ligation causes endothelial dysfunction in a week, followed by robust and rapid atheroma formation in two weeks in a hyperlipidemic mouse model along with features of complex lesion formation such as intraplaque neovascularization by four weeks. This rapid in vivo model and the endothelial RNA preparation method could be used to determine molecular mechanisms underlying flow-dependent regulation of vascular biology and diseases. Also, it could be used to test various therapeutic interventions targeting endothelial dysfunction and atherosclerosis in considerably reduced study duration.

This describes a partial carotid ligation surgery, which causes disturbed flow conditions and subsequent atherosclerosis development (in two weeks) with intraplaque neo-vascularization (in four weeks) in the mouse common carotid artery. We also describe a novel method of RNA isolation from the carotid intima, providing high purity endothelial RNA.

部分的ライゲーションによるマウス頸動脈における乱流誘発アテローム性動脈硬化症と頸動脈内皮からのRNA単離の方法における簡便な方法のモデル

Despite the well-known close association, direct evidence linking disturbed flow to atherogenesis has been lacking. We have recently used a modified ...

医学

Murine Spinotrapezius Model to Assess the Impact of Arteriolar Ligation on Microvascular Function and Remodeling

The murine spinotrapezius is a thin, superficial skeletal support muscle that extends from T3 to L4, and is easily accessible via dorsal skin incision. Its unique anatomy makes the spinotrapezius useful for investigation of ischemic injury and subsequent microvascular remodeling. Here, we demonstrate an arteriolar ligation model in the murine spinotrapezius muscle that was developed by our research team and previously published1-3. For certain vulnerable mouse strains, such as the Balb/c mouse, this ligation surgery reliably creates skeletal muscle ischemia and serves as a platform for investigating therapies that stimulate revascularization. Methods of assessment are also demonstrated, including the use of intravital and confocal microscopy. The spinotrapezius is well suited to such imaging studies due to its accessibility (superficial dorsal anatomy) and relative thinness (60-200 &mu;m). The spinotrapezius muscle can be mounted en face, facilitating imaging of whole-muscle microvascular networks without histological sectioning. We describe the use of intravital microscopy to acquire metrics following a functional vasodilation procedure; specifically, the increase in arterilar diameter as a result of muscle contraction. We also demonstrate the procedures for harvesting and fixing the tissues, a necessary precursor to immunostaining studies and the use of confocal microscopy.

We demonstrate a novel arterial ligation model in murine spinotrapezius muscle, including a step-by-step procedure and description of required instrumentation. We describe the surgery and relevant outcome measurements relating to vascular network remodeling and functional vasodilation using intravital and confocal microscopy.

微小血管機能およびリモデリングに動脈結紮の影響を評価するためにネズミSpinotrapeziusモデル

The murine spinotrapezius is a thin, superficial skeletal support muscle that extends from T3 to L4, and is easily accessible via dorsal skin incision. ...

A Murine Model of Stent Implantation in the Carotid Artery for the Study of Restenosis

Despite the considerable progress made in the stent development in the last decades, cardiovascular diseases remain the main cause of death in western countries. Beside the benefits offered by the development of different drug-eluting stents, the coronary revascularization bears also the life-threatening risks of in-stent thrombosis and restenosis. Research on new therapeutic strategies is impaired by the lack of appropriate methods to study stent implantation and restenosis processes. Here, we describe a rapid and accessible procedure of stent implantation in mouse carotid artery, which offers the possibility to study in a convenient way the molecular mechanisms of vessel remodeling and the effects of different drug coatings.

A model of stent implantation in mouse carotid artery is described. Compared to other similar methods, this procedure is very rapid, simple and accessible, offering the possibility to study in a convenient way the vascular wall reaction to different drug-eluting stents and the molecular mechanisms of restenosis.

再狭窄の研究のための頸動脈内ステント留置のマウスモデル

Despite  the considerable progress made in the stent development in the last decades,  cardiovascular diseases remain the main cause of death in western ...

Vascular Balloon Injury and Intraluminal Administration in Rat Carotid Artery

The carotid artery balloon injury model in rats has been well established for over two decades. It remains an important method to study the molecular and cellular mechanisms involved in vascular smooth muscle dedifferentiation, neointima formation and vascular remodeling. Male Sprague-Dawley rats are the most frequently employed animals for this model. Female rats are not preferred as female hormones are protective against vascular diseases and thus introduce a variation into this procedure. The left carotid is typically injured with the right carotid serving as a negative control. Left carotid injury is caused by the inflated balloon that denudes the endothelium and distends the vessel wall. Following injury, potential therapeutic strategies such as the use of pharmacological compounds and either gene or shRNA transfer can be evaluated. Typically for gene or shRNA transfer, the injured section of the vessel lumen is locally transduced for 30 min with viral particles encoding either a protein or shRNA for delivery and expression in the injured vessel wall. Neointimal thickening representing proliferative vascular smooth muscle cells usually peaks at 2 weeks after injury. Vessels are mostly harvested at this time point for cellular and molecular analysis of cell signaling pathways as well as gene and protein expression. Vessels can also be harvested at earlier time points to determine the onset of expression and/or activation of a specific protein or pathway, depending on the experimental aims intended. Vessels can be characterized and evaluated using histological staining, immunohistochemistry, protein/mRNA assays, and activity assays. The intact right carotid artery from the same animal is an ideal internal control. Injury-induced changes in molecular and cellular parameters can be evaluated by comparing the injured artery to the internal right control artery. Likewise, therapeutic modalities can be evaluated by comparing the injured and treated artery to the control injured only artery.

This protocol uses a balloon catheter to cause an intraluminal injury on the rat carotid artery and henceforth elicit neointimal hyperplasia. This is a well-established model for studying the mechanisms of vascular remodeling in response to injury. It is also widely used to determine the validity of potential therapeutic approaches.

ラット頸動脈における血管バルーン傷害と管腔内投与

The carotid artery balloon injury model in rats has been well established for over two decades. It remains an important method to study the molecular and ...

Murine Model of Femoral Artery Wire Injury with Implantation of a Perivascular Drug Delivery Patch

Percutaneous interventions including balloon angioplasty and stenting have been used to restore blood flow in vessels with occlusive vascular disease. While these therapies lead to the rapid restoration of blood flow, these technologies remain limited by restenosis in the case of bare metal stents and angioplasty, or reduced healing and possibly enhanced risk of thrombosis in the case of drug eluting stents. A key pathophysiological mechanism in the formation of restenosis is intimal hyperplasia caused by the activation of vascular smooth muscle cells and inflammation due to arterial stretch and injury. Surgeries that induce arterial injury in genetically modified mice are useful for the mechanistic study of the vascular response to injury but are often technically challenging to perform in mouse models due to the their small size and lack of appropriate sized devices. We describe two approaches for a surgical technique that induces endothelial denudation and arterial stretch in the femoral artery of mice to produce robust neointimal hyperplasia. The first approach creates an arteriotomy in the muscular branch of the femoral artery to obtain vascular access. Following wire injury this arterial branch is ligated to close the arteriotomy. A second approach creates an arteriotomy in the main femoral artery that is later closed through localized cautery. This method allows for vascular access through a larger vessel and, consequently, provides a less technically demanding procedure that can be used in smaller mice. Following either method of arterial injury, a degradable drug delivery patch can be placed over or around the injured artery to deliver therapeutic agents.

We describe a surgical technique that produces wire injury in the femoral artery of mice to induce neointimal hyperplasia to serve as a model testing system for the perivascular delivery of therapeutic compounds for the inhibition of restenosis.

血管周囲ドラッグデリバリーパッチの移植に大腿動脈ワイヤ傷害のマウスモデル

Percutaneous interventions including balloon angioplasty and stenting have been used to restore blood flow in vessels with occlusive vascular disease. ...

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. Moreover, by rupture of the defective vascular wall, atherosclerosis induces occlusive thrombus formation, which represents the main cause of myocardial infarction or stroke and the most frequent cause of death. Despite the advances in the cardiovascular field, many questions remain unanswered, and additional basic research is essential to improve our understanding of the molecular mechanisms during atherosclerosis and its effects. Due to limited clinical studies, there is a need for representative animal models recreating atherosclerotic conditions such as neointima formation after stent implantation, balloon angioplasty, or endarterectomy. Since the mouse presents many advantages and is the most frequently used model for studying molecular processes, the current study proposes an invasive procedure of endothelial denudation, also known as the wire-injury model, which is representative of the human condition of neointima formation in arteries after revascularization procedures.

This study describes an invasive procedure for the induction of accelerated atherosclerosis in mice. In comparison to other methods using electric- or cryo-induced injury, mechanical-induced injury mimics the human condition of restenosis after revascularization therapies and is ideal for the study of the molecular mechanisms involved.

マウスにおける加速アテローム性動脈硬化症の誘導:ワイヤー傷害モデル

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...

Reproducible Arterial Denudation Injury by Infrarenal Abdominal Aortic Clamping in a Murine Model

Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These complications can be attributed to impairments in re-endothelialization within the wound margins. Yet, the cellular and molecular mechanisms of re-endothelialization remain to be defined. While several animal models to study re-endothelialization after arterial denudation are available, few are performed in the mouse because of surgical limitations. This undermines the opportunity to exploit transgenic mouse lines and investigate the contribution of specific genes to the process of re-endothelialization. Here, we present a step-by-step protocol for creating a highly reproducible murine model of arterial denudation injury in the infrarenal abdominal aorta using external vascular clamping. Immunocytochemical staining of injured aortas for fibrinogen and &#946;-catenin demonstrate the exposure of a pro-thrombotic surface and the border of intact endothelium, respectively. The method presented here has the advantages of speed, excellent overall survival rate, and relative technical ease, creating a uniquely practical tool for imposing arterial denudation injury in transgenic mouse models. Using this method, investigators may elucidate the mechanisms of re-endothelialization under normal or pathological conditions.

Understanding the cellular and molecular mechanisms of re-endothelialization following arterial denudation injury is of paramount importance in preventing thrombosis and restenosis of arteries. Here we describe a protocol for reproducible arterial denudation injury of the infrarenal abdominal aorta. The procedure was developed to investigate the underlying mechanisms that regulate endothelial regeneration using mouse models.

マウスモデルにおける下腹部大動脈クランプすることにより、再現性動脈侵食に起因けが

Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These ...

The Rabbit Model of Accelerated Atherosclerosis: A Methodological Perspective of the Iliac Artery Balloon Injury

Acute coronary syndrome resulting from coronary occlusion following atherosclerotic plaque development and rupture is the leading cause of death in the industrialized world. New Zealand White (NZW) rabbits are widely used as an animal model for the study of atherosclerosis. They develop spontaneous lesions when fed with atherogenic diet; however, this requires long time of 4 - 8 months. To further enhance and accelerate atherogenesis, a combination of atherogenic diet and mechanical endothelial injury is often employed. The presented procedure for inducing atherosclerotic plaques in rabbits uses a balloon catheter to disrupt the endothelium in the left iliac artery of NZW rabbits fed with atherogenic diet. Such mechanical damage caused by the balloon catheter induces a chain of inflammatory reactions initiating neointimal lipid accumulation in a time dependent fashion. Atherosclerotic plaque following balloon injury show neointimal thickening with extensive lipid infiltration, high smooth muscle cell content and presence of macrophage derived foam cells. This technique is simple, reproducible and produces plaque of controlled length within the iliac artery. The whole procedure is completed within 20 - 30 min. The procedure is safe with low mortality and also offers high success in obtaining substantial intimal lesions. The procedure of balloon catheter induced arterial injury results in atherosclerosis within two weeks. This model can be used for investigating the disease pathology, diagnostic imaging and to evaluate new therapeutic strategies.

Animal models of atherosclerosis are essential to understand the mechanism and to investigate newer approaches to prevent plaque development or rupture, a leading cause of death in the industrialized world. This protocol uses a combination of balloon injury and cholesterol rich diet to induce atherosclerotic plaques in rabbit iliac artery.

加速動脈硬化家兎モデル: 内腸骨動脈バルーン傷害の方法論的視点

Acute coronary syndrome resulting from coronary occlusion following atherosclerotic plaque development and rupture is the leading cause of death in the ...

A Rat Carotid Artery Pressure-Controlled Segmental Balloon Injury with Periadventitial Therapeutic Application

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the luminal surface area in arteries thereby reducing adequate blood flow to organs and distal tissues. Clinically, revascularization procedures such as balloon angioplasty with or without stent placement aim to restore blood flow. Although these procedures reestablish blood flow by reducing plaque burden, they damage the vessel wall, which initiates the arterial healing response. The prolonged healing response causes arterial restenosis, or re-narrowing, ultimately limiting the long-term success of these revascularization procedures. Therefore, preclinical animal models are integral for analyzing the pathophysiological mechanisms driving restenosis, and provide the opportunity to test novel therapeutic strategies. Murine models are cheaper and easier to operate on than large animal models. Balloon or wire injury are the two commonly accepted injury modalities used in murine models. Balloon injury models in particular mimic the clinical angioplasty procedure and cause adequate damage to the artery for the development of restenosis. Herein we describe the surgical details for performing and histologically analyzing the modified, pressure-controlled rat carotid artery balloon injury model. Additionally, this protocol highlights how local periadventitial application of therapeutics can be used to inhibit neointimal hyperplasia. Lastly, we present light sheet fluorescence microscopy as a novel approach for imaging and visualizing the arterial injury in three-dimensions.

The rat carotid artery balloon injury mimics the clinical angioplasty procedure performed to restore blood flow in atherosclerotic vessels. This model induces the arterial injury response by distending the arterial wall, and denuding the intimal layer of endothelial cells, ultimately causing remodeling and an intimal hyperplastic response.

過期治療アプリケーションを用いてラット頸動脈圧制御区分バルーン損傷

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the ...

Murine Aortic Crush Injury: An Efficient In Vivo Model of Smooth Muscle Cell Proliferation and Endothelial Function

Arterial reconstruction, whether angioplasty or bypass surgery, involves iatrogenic trauma causing endothelial disruption and vascular smooth muscle cell (VSMC) proliferation. Common murine models study small vessels such as the carotid and femoral arteries. Herein we describe an in vivo system in which both VSMC proliferation and endothelial barrier function can be simultaneously assessed in a large vessel. We studied the infrarenal aortic response to injury in C57BL/6 mice. The aorta was injured from the left renal vein to the aortic bifurcation by 30 transmural crushes of 5-seconds duration with a cotton-tipped applicator. Morphological changes were assessed with conventional histology. Aorta wall thickness was measured from the luminal surface to the adventitia. EdU integration and counter staining with DAPI and alpha-actin was used to demonstrate VSMC proliferation. Activation of ERK1/2, a known moderator of intimal hyperplasia formation, was determined by Western Blot analysis. The effect of inflammation was determined by immunohistochemistry for B-cells, T-cells, and macrophages. En face sections of endothelium were visualized with scanning electron microscopy (SEM). Endothelial barrier function was determined with Evans Blue staining. Transmural injury resulted in aortic wall thickening. This injury induced VSMC proliferation, most prominently at 3 days after injury, and early activation of ERK1/2 and decreased p27kip1 expression. Injury did not result in increased B-cells, T-cells, or macrophages infiltration in the vessel wall. Injury caused partial endothelial cell denudation and loss of cell-cell contact. Injury resulted in a significant loss of endothelial barrier function, which returned to baseline after seven days. The murine transmural blunt aortic injury model provides an efficient system to simultaneously study both VSMC proliferation and endothelial barrier function in a large vessel.

Murine Aortic Crush Injury: An Efficient In Vivo Model of Smooth Muscle Cell Proliferation and Endothelial Function

Restenosis following cardiovascular procedures (bypass surgery, angioplasty, or stenting) is a significant problem reducing the durability of these procedures. An ideal therapy would inhibit smooth muscle cell (VSMC) proliferation while promoting regeneration of the endothelium. We describe a model for simultaneous assessment of VSMC proliferation and endothelial function in vivo.

ネズミ大動脈挫傷：効率的<em&gt;インビボ</em&gt;平滑筋細胞の増殖と内皮機​​能のモデル

Arterial reconstruction, whether angioplasty or bypass surgery, involves iatrogenic trauma causing endothelial disruption and vascular smooth muscle cell ...