Name: murine left anterior descending (lad) coronary artery ligation: an improved and simplified model for myocardial infarction
Uploaded: 2017-04-02T15:00:00.000+00:00
Duration: 10 min 39 s
Description: Watch this Scientific Journal Video about Murine Left Anterior Descending LAD Coronary Artery Ligation: An Improved and Simplified Model for Myocardial Infarction at JoVE.com

This model was developed with the support of the National Institute of General Medical Sciences (NIGMS)/the National Institute of Health (NIH) grant 1P20GM103652 (Project# 3) (to MRA) and the American Heart Association (AHA) Grant-in-Aid 14GRNT20460291 (to MRA); the Brazilian government grant CAPES (to KR and FR); and a Brown University LINK award (to IM). We also acknowledge the outstanding technical support from our veterinarians and animal facility staff.

Bu hayvan protokolü gözden geçirmiş ve Rhode Island Hastanesinde Kurumsal Hayvan Bakım ve Kullanım Komitesi (IACUC) tarafından onaylanmıştır. 1. Anestezi ve Entübasyon <ol><li> ameliyat sonrası ağrı ilaç dozunu hesaplamak için fare tartılır. </li><li> indüksiyon odasında fare yerleştirin ve 9 için% 4 izofluran teslim - boyunca hayvan izleme, 10 dakika. Cihaz, yaklaşık 250 ° C&#39;ye kadar ön ısıtma, böylece sıcak bir boncuk sterilizatör açın. 20 dakika - Ön ısıtma 15 alacaktır. </li><li> Fare yaklaşık 32 nefes / dk bir nefes oranı ile anestezi derinliğinin uçağı, ulaştığında, bir strafor gemide fare sırtüstü yatırın yerleştirin ve açık ağzını tutmak için üst kesici dişler güvence altına elastik bir bant kullanın. Bir ayak tutam yaparak sedasyon onaylayın. orofarenks görüntülenmiştir böylece fare üzerinde yüksek yoğunluklu bir ışığı yerleştirin. </li><li> çene ve bir çift açık eğri forsepsforseps dışına dilini kaldırmaya karar verdi. veya biraz fare vücuduyla göz seviyesinin altında konumlandırılmış ise entübe emin olun. Cerrahi loupes kullanılması tavsiye edilir. </li><li> ses telleri açılıp kapanmasını Görselleştirin. Açıkken eklemek 20 gauge, 1 in bir küt uçlu bir iğne introduser intravenöz (IV) kateter. trakea açılışına kılavuz kateterin ama trakea içine iğneyle önlemek için iğne kullanın. Doğru yerleştirme Doğrulama plastik bir transfer pipet kullanarak yapılabilir. </li><li> Bir ısıtma cihazı ile donatılmış bir çalışma yüzeyine entübe fare aktarın. 150 uL / ​​inme felç hacmi ve / dakika 130 vuruş bir hareket hızına ayarlanmış küçük bir kemirgen vantilatöre fare bağlayın. </li><li> % 2.5 izofluran verin. ikili göğüs yükselişi kontrol ederek entübasyon olun. Bir ayak tutam yaparak anestezi olun. Tam anestezi olmak üzere vantilatör 10 dk - fare 5 gerekebilir. </li></ol><ol><li> Vantilatör ve IV kateter arasındaki bağlantı yerinde entübasyon aşağı tüp bant. ekstremitelerde aşağı Teyp. Gözleri steril yağlama damla yerleştirin. </li><li> Bir elektrik ustura ile toraks ventral sol tarafını Trim. Kuru mendil ile tıraş kürk toz ve steril bir pamuk bez kullanılarak epilasyon kremi küçük bir tabaka uygulanır. 45 sn - krem ​​yaklaşık 30 saç folikülleri ile temas halinde kalmalıdır. </li><li> krem işlemleri sırasında, emmek için Betadine dolu üç 1.5 mL tüpler üç steril pamuk ile yerleştirin. damıtılmış su ile nemlendirilmiş olan mendiller kullanarak, yavaşça krema ve kürk silin. </li><li> merkezden çevreye doğru hareket eden bir dairesel hareket içinde, temizleme Betadine ve steril% 70 izopropanol hazırlık pedleri alternatif, cerrahi alan üç kez temizleyin. fare cerrahi alanın üzerine çeyrek ölçülü delik ile bir steril örtüyü yerleştirin. </li><li> mous çevresindeki alanı temizleyin% 70 etanol ile, e. bir kez daha bir ayak tutam anestezi olun. </li></ol> 3. LAD Ligasyon <ol><li> yaklaşık 20 saniye boyunca 250 ° C&#39;ye ısıtılmış, sıcak bir boncuk sterilizatör otoklava cerrahi aletleri yerleştirin. Steril otoklavlanmış cerrahi örtü üzerine steril aletleri yerleştirin. Don ameliyat eldivenleri. </li><li> nazikçe belirgin kılıç şeklinde kıkırdak solunda bir noktada yaklaşık 5 mm deriyi kaldırmak için ince uçlu forseps kullanın. manubrium seviyesine yukarı bu noktadan deride dikey kesi oluşturmak için bir No. 10 bıçak ile bir cerrahi neşter kullanın. </li><li> yumuşak kas ve deri tabakaları ayırmak için eğri forseps. cilt kesisi sonrasında kas tabakası açın. kas tabakasının, kesi her iki tarafında biri aracılığıyla iki 5-0 polipropilen sütür yerleştirin ve kas tabakası açık tutmak için kelepçeleri ile geçici olarak Sütürleri. </li><li> Aşağıdaki, tanımlama ve üçüncü interkostal bir kesi yapmakgöğüs kafesi doğal açısı. farenin sol ekstremite bandı çıkarın ve bantla sağ arka ayağı olan sol arka ayağını sabitleyin. Bandın uzun parça kesin ve hafifçe yükseltilmiş pozisyonda ameliyat yüzeyine sol ön ayağı sabitleyen. % 70 etanol ile eldiven temizleyin. </li><li> Nazikçe 3. ve 4. kaburga ayırın Retraktörün kullanın. yaklaşık olarak 1 x koklea steril bir gazlı bez küçük bir bölümü kesin ve steril% 0.9 tuzlu su içinde daldırma. aşırı tuzlu su sıkın ve yavaşça işlem sırasında kazara akciğer hasarı önlemek için sol akciğer karşı gazlı eklemek için forseps kullanır. </li><li> Yavaşça forseps ile ince perikard çıkarın. </li><li> Steril bir pamuklu çubukla kapalı pamuk az miktarda Gözyaşı ve küçük top şekline getirin. steril% 0.9 tuzlu su içine bu pamuk Dip ve yavaşça arterleri takdir kalp yüzeyi üzerinde bez. Yavaşça yukarıya doğru sol kulak kepçesi itip altında koroner arterler bulunneath. </li><li> LAD belirlenmesi ve LAD altına bir 8-0 naylon dikiş geçmesi; İki tam ligasyonu sabitlemek için atar. ligasyonu başarılı olursa, bağ irade blanch sol ventrikül uzak. </li><li> forseps kullanarak, daha önce yerleştirilen tül kaldırmak ve sonra yavaşça Retraktörü çıkarın. torakotomi açıklığından göğüs boşluğuna 25-gauge iğne takılı bir 6-in-25-gauge, esnek tüp yerleştirin. sol akciğer üzerindeki boşluk içine boru 2 - yaklaşık 1 Advance. Bir yatar pozisyonda fare dönün ve% 70 etanol ile eldiven temizleyin. </li><li> yerinde göğüs tüpü tutarak, göğüs kafesi kapatmak için basit Kesintiye desen 5-0 polipropilen sütür kullanın. Açık kas tabakasını tutan iki dikiş çıkarın. yerinde tekrar göğüs tüpü tutmak, kas tabakası kapatmak için basit bir sürekli olan bir model içinde 5-0 polipropilen dikişlerle kullanın. </li><li> göğüs tüpü 25 numara bir iğnenin bir 1 ml şırınga takın. Yavaşça dalma ile yukarı doğru çekerekr aynı anda yavaş yavaş forseps ile göğüs boşluğundan göğüs tüpü çakılırken. Bu adım, başka bir şekilde pnömotoraks göğüs boşluğunda ve sonuç olarak takılıp kalabilmektedir fazla havayı ve kan kaldırır, yavaş yavaş boru ekstrakte edin. </li><li> Şırınga dolduğunda, iğne, şırınganın ayırmak ve bir atık cam kap veya lavaboya atık imha. göğüs tüpü tamamen çekme kadar bu işleme devam edin. göğüs sıkıca kapatılmış olduğundan emin olun. </li><li> % 1.5 izofluran azaltır. Basit bir Kesintiye desen 4-0 polipropilen sütür ile cilt kapatın. kapalı izofluran buharlaştırıcı açın. </li><li> intraperitonal (İP) enjeksiyon yoluyla% 0.9 tuzlu su içinde 0.1 mg / mL buprenorfin yönetmek. Topikal kesi% 0.9 tuzlu su içinde 2 mg / mL Bupivakain 2 mg / ml lidokain uygulanır. Fare ağırlığına tuz miktarı ölçekleme, bir deri altından enjeksiyon ile% 0.9 salin 500 uL - 200 arasında uygulayınız. </li><li> Admini sonra 5 dakika bekleyinağrı kesici stering entübasyon tüpten fare kaldırın. Bu solunumunu durdurma geçişte yardımcı olur. <ol><li> Fare solunumunu durdurma kez ikili göğüs artış yoksa, iğne açmayı gerçekleştirmektedir. Bu direncinde ani azalma ile belirtilen göğüs boşluğu, girene kadar Bunu yapmak için, 25-gauge, steril bir iğne ve 3 ve 4. kaburgalar arasındaki bir 1 ml hava tanıtır. Fazla havayı çıkarmak için pistonu yavaşça yukarı doğru çekin. </li></ol></li><li> Fare yeterli ikili solunum hızı ve derinliği gösterir ve bir ayak tutam yanıt verdiğinde, bir ısı lambası altında temiz bir kurtarma kafesine fareyi yerleştirin. 20 dakika - 15, bir laminar akış başlığı içinde izlenmesi, nemli gıda ve su şişesi ile fare sağlar. abartılı bir nefes çabası, aşırı kanama veya diğer potansiyel olarak yaşamı tehdit eden komplikasyonlar için izleyin. </li><li> Üç gün için, bir IP vasıtasıyla 0.1 mg / mL buprenorfin ağrı ilaç yönetmekGünde iki kez jection. Günlük fare izleyin. </li></ol>

<ol>
	<li>Hausenloy, D. J., Yellon, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+directions+for+protecting+the+heart+against+ischaemia-reperfusion+injury:+targeting+the+Reperfusion+Injury+Salvage+Kinase+(RISK)-pathway.">New directions for protecting the heart against ischaemia-reperfusion injury: targeting the Reperfusion Injury Salvage Kinase (RISK)-pathway.</a> Cardiovasc Res. 61 (3), 448-460 (2004).</li><li>Roffi, 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=2015+ESC+Guidelines+for+the+management+of+acute+coronary+syndromes+in+patients+presenting+without+persistent+ST-segment+elevation.">2015 ESC Guidelines for the management of acute coronary syndromes in patients presenting without persistent ST-segment elevation.</a> Eur Heart J. 37 (3), 267-315 (2015).</li><li>Kumar, A., Cannon, C. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+Coronary+Syndromes:+Diagnosis+and+Management,+Part+I.">Acute Coronary Syndromes: Diagnosis and Management, Part I.</a> Mayo Clin Proc. 84 (10), 917-938 (2009).</li><li>Eitan, A., Nikolsky, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antithrombotic+therapy+in+patients+with+acute+coronary+syndromes:+how+to+make+the+right+choice.">Antithrombotic therapy in patients with acute coronary syndromes: how to make the right choice.</a> Minerva Med. 104 (4), 357-381 (2013).</li><li>Abbate, A., Bussani, R., Amin, M. S., Vetrovec, G. W., Baldi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+myocardial+infarction+and+heart+failure:+role+of+apoptosis.">Acute myocardial infarction and heart failure: role of apoptosis.</a> Int J Biochem Cell Biol. 38 (11), 1834-1840 (2006).</li><li>Zheng, 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=Nebivolol+protects+against+myocardial+infarction+injury+via+stimulation+of+beta+3-adrenergic+receptors+and+nitric+oxide+signaling.">Nebivolol protects against myocardial infarction injury via stimulation of beta 3-adrenergic receptors and nitric oxide signaling.</a> PLOS ONE. 9 (5), 98179 (2014).</li><li>Tarnavski, O., 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=Mouse+cardiac+surgery:+comprehensive+techniques+for+the+generation+of+mouse+models+of+human+diseases+and+their+application+for+genomic+studies.">Mouse cardiac surgery: comprehensive techniques for the generation of mouse models of human diseases and their application for genomic studies.</a> Physiol Genomics. 16 (3), 349-360 (2004).</li><li>Buitrago, S., Martin, T. E., Tetens-Woodring, J., Belicha-Villanueva, A., Wilding, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Safety+and+Efficacy+of+Various+Combinations+of+Injectable+Anesthetics+in+BALB/c+Mice.">Safety and Efficacy of Various Combinations of Injectable Anesthetics in BALB/c Mice.</a> J Am Assoc Lab Anim Sci. 47 (1), 11-17 (2008).</li><li>Kolk, M. V., 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=LAD-Ligation:+A+Murine+Model+of+Myocardial+Infarction.">LAD-Ligation: A Murine Model of Myocardial Infarction.</a> J Vis Exp. (32), e1438 (2009).</li><li>Wu, Y., Yin, X., Wijaya, C., Huang, M. H., McConnell, B. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+myocardial+infarction+in+rats.">Acute myocardial infarction in rats.</a> Journal of visualized experiments : J Vis Exp. (48), (2011).</li><li>Ferrera, R., Benhabbouche, S., Bopassa, J. C., Li, B., Ovize, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=One+hour+reperfusion+is+enough+to+assess+function+and+infarct+size+with+TTC+staining+in+Langendorff+rat+model.">One hour reperfusion is enough to assess function and infarct size with TTC staining in Langendorff rat model.</a> Cardiovasc Drugs Ther. 23 (4), 327-331 (2009).</li><li>Zeng, 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=Evaluation+of+5-ethynyl-2'-deoxyuridine+staining+as+a+sensitive+and+reliable+method+for+studying+cell+proliferation+in+the+adult+nervous+system.">Evaluation of 5-ethynyl-2'-deoxyuridine staining as a sensitive and reliable method for studying cell proliferation in the adult nervous system.</a> Brain Res. 1319, 21-32 (2010).</li></ol>

The authors declare that they have no competing financial interests.

Laboratuvar MI modelinin artan kullanımıyla birlikte, tarif edilen prosedürü, ameliyat sonrası ağrı ve rahatsızlık en aza indirirken, farelerin verim ve hayatta kalma oranını geliştirmek için çalışmaktadır. Bu protokol, LAD ligasyonu prosedürün çeşitli yönlerine çeşitli iyileştirmeler yaparak mortalite en aza indirmek için çaba göstermektedir. Birkaç ayrımlar vardır. Nedeniyle anestezi bunların uzun süreli yararına, indüksiyon için izofluran ile birlikte ketamini ve k...

Koroner hastalık veya ACS, en yaygın kardiyovasküler olay ve 2020 1&#39;de morbidite ve mortalitenin dünya çapında ana nedeni olarak kabul edilecektir. ACS nedeni nedeniyle örneğin kalp dokusu 2 kan akışını azaltan bloke eden veya bir koroner aterosklerotik plak yırtılması için bir miyokardiyal tromboz varlığıdır. Bu nedenle, miyokardiyal enfarktüs (MI) 3, 4, akut miyokart iskemisi mevcudiyeti ile uyumlu klinik işaretler vardır. MI ventriküler disfonksiyon ve kalp yetmezliği 5, 6 yol açabilir kardiyomiyositlerinin kütle kaybı ve patolojik ventriküler yeniden bir ilerleme, yol açar. IHD çalışma için en etkin yollarından biri, bir hayvan modelinde insan miyokardiyal enfarktüs taklit etmek olmuştur. Bu LAD içinde tıkayarak elde edilirFareler. Bu modeli kullanarak, kalp İHD kaynaklanan hasarlara karşı korunabilir nasıl inceliyoruz. Son on yılda, araştırmacılar farelere sıçanlardan kayması dahil, daha küçük hayvanlara büyük hayvan modelleri kullanılarak kaymasıyla. Daha küçük fare modeli, küçük boyutları dahil olmak üzere çeşitli nedenlerle, büyük çöp boyutu korumak için düşük maliyetli ve kısa gebelik süresi, hem de transjenik ve gen nakavt modeller 7 geniş kullanılabilirliği için tercih başlıyor. Fareler boyutu küçük olmasına rağmen, kendilerine özel geliştirilen yeni cerrahi aletleri bu gelişmede yardımcı olmaktadır. Bizim yöntem bu yeni cerrahi araçlar kullanmaktadır. çeşitli yöntemler invaziv bir trakeotomi uygulamak iken, biz endotrakeal entübasyon daha az invazif yöntemi kullanın. Orofarenksin havai aydınlatma kullanarak, t için daha güvenli ve daha az travmatik bir deneyim sağlayan, herhangi bir kesi oluşturmadan entübeo hayvan. Daha sonra fare bir vantilatör yerleştirilir ve tüm işlem boyunca izofluran tutulur. o kesildiğinde hayvan anestezi kurtarmak için nedeniyle ilaç tarafından üretilen anestezi kısa süreli olmasından, sadece birkaç dakika sürer. Bizim cerrahi modeli de minimal invaziv torasenteze içerir. germe pnömotoraks: Orijinal torakotomi kesikten thoracentesis kullanılarak göğüs boşluğundan kan ve fazla hava dikkatlice çıkarılması, LAD ligasyon ortak bir ameliyat sonrası komplikasyonlar ele almıştır. Diğer yöntemler-on torasentezler-etti vermiştir az ameliyat sonrası komplikasyonlar için trakeotomi ve başka kullanılan iki ilave yarıkların olan ihtiyacı ortadan kaldırır ve büyük ölçüde ölüm oranı azalmış olmakla bu yöntem.

<table><tbody><tr><td>High-Intensity Light Source</td><td>Harvard Apparatus</td><td>72-0215</td><td></td></tr><tr><td>SurgiSuite Operating Platform</td><td>Kent Scientific Corporation</td><td>SurgiSuite</td><td>Uses a rechargeable, battery-operated far infrared warming pad. Charge overnight before surgery.&amp;nbsp;</td></tr><tr><td>SurgiSuite LED Lighting Kit</td><td>Kent Scientific Corporation</td><td>SURGI-5003</td><td></td></tr><tr><td>Hot Bead Sterilizer</td><td>Fine Science Tools</td><td>18000-45</td><td>Preheating takes 15 - 20 min. Instruments take 20 s to sterilize.</td></tr><tr><td>Small Rodent Anesthesia System</td><td>VetEquip Inc.</td><td>901810</td><td></td></tr><tr><td>Isofluorane</td><td>Piramal Enterprises</td><td>66794-017-10</td><td></td></tr><tr><td>Buprenorphine</td><td>Rhode Island Hospital Pharmacy</td><td>NDC 12496-0757-1, 12496-0757-5</td><td></td></tr><tr><td>Surgical Loupes</td><td>Roboz</td><td>RS-6687</td><td></td></tr><tr><td>Small Rodent Ventilator</td><td>Harvard Apparatus</td><td>73-0043</td><td></td></tr><tr><td>Lubricating Drops</td><td>Thermo Fisher Scientific</td><td>19-898-350</td><td></td></tr><tr><td>Electric Razor</td><td>Kent Scientific Corporation</td><td>CL 9990-1201</td><td></td></tr><tr><td>Hair Removal Cream</td><td>Nair</td><td></td><td></td></tr><tr><td>Medical Tape</td><td>Thermo Fisher Scientific</td><td>18-999-380</td><td></td></tr><tr><td>Betadine</td><td>Thermo Fisher Scientific</td><td>19-027136</td><td></td></tr><tr><td>70% Isopropanol Wipes</td><td>Thermo Fisher Scientific</td><td>22-363-750</td><td></td></tr><tr><td>Surgical Drapes</td><td>Braintree</td><td>SP-TS</td><td></td></tr><tr><td>Surgical Gloves</td><td>Thermo Fisher Scientific</td><td>18999102D</td><td></td></tr><tr><td>5-0 Polypropylene Sutures&amp;nbsp;</td><td>Ethicon</td><td>8630G</td><td></td></tr><tr><td>8-0 Nylon Sutures</td><td>Fine Science Tools</td><td>12051-08</td><td></td></tr><tr><td>Platinum-Cured Tubing</td><td>Harvard Apparatus</td><td>72-1042&amp;nbsp;</td><td>0.3 mm inside diameter x 0.6 mm outside diameter</td></tr><tr><td>0.9% Saline</td><td>Thermo Fisher Scientific</td><td>19-310-207</td><td></td></tr><tr><td>4-0 Polypropylene Sutures</td><td>Ethicon</td><td>8631G</td><td></td></tr><tr><td>1 CC Syringe with 25-Gauge Needle&amp;nbsp;</td><td>Thermo Fisher Scientific</td><td>14-826-100</td><td></td></tr><tr><td>Scissors</td><td>Kent Scientific Corporation</td><td>INSS600225</td><td></td></tr><tr><td>Forceps</td><td>Kent Scientific Corporation</td><td>INS700100</td><td></td></tr><tr><td>Cotton Swabs</td><td>Thermo Fisher Scientific</td><td>23-400-118</td><td></td></tr><tr><td>IV Catheter, 20-Gauge</td><td>Thermo Fisher Scientific</td><td>&amp;nbsp;NC9892181</td><td></td></tr><tr><td>Retractor</td><td>Kent Scientific Corporation</td><td>INS 750369</td><td></td></tr><tr><td>Forceps</td><td>Fine Science Tools</td><td>11003-12</td><td></td></tr><tr><td>Dissecting Forceps, Straight</td><td>Kent Scientific Corporation</td><td>INS 700101</td><td></td></tr><tr><td>Dissecting Forceps, Curved</td><td>Kent Scientific Corporation</td><td>INS 700103</td><td></td></tr><tr><td>Hemostatic Forceps, Straight</td><td>Kent Scientific Corporation</td><td>INS 750451</td><td></td></tr><tr><td>Hemostatic Forceps, Curved</td><td>Kent Scientific Corporation</td><td>INS 750452</td><td></td></tr><tr><td>Tissue Forceps</td><td>Kent Scientific Corporation</td><td>INS 700131</td><td></td></tr><tr><td>Needle Holder</td><td>Kent Scientific Corporation</td><td>INS 600109</td><td></td></tr><tr><td>Scissors&amp;nbsp;</td><td>Kent Scientific Corporation</td><td>INS 600225</td><td></td></tr></tbody></table>

murine left anterior descending (lad) coronary artery ligation: an improved and simplified model for myocardial infarction

İskemik kalp hastalığı (İHD), veya akut koroner sendrom (AKS), Amerika Birleşik Devletleri&#39;nde önde gelen ölüm nedenlerinden biridir. IHD oksijen kaybına ve kalp kası takip eden nekroz ile sonuçlanan, kalp Kan akışında ile karakterize edilir. MI modeli kısa süreli iskemi-reperfüzyon modeli ve uzun vadeli kalıcı ligasyonu model olarak kullanılmak üzere popülerlik kazanmıştır. Aşağıda, LAD kalıcı ligasyonu için güvenilir bir yöntem açıklanmaktadır. Fare genetik mühendisliği teknolojisi daha gelişmiş hale geliyor ve kaliteli fare cerrahi aletlerin artan kullanılabilirliği ile birlikte, fare MI ameliyatları için popüler bir model haline gelmiştir. Bizim cerrahi modeli fare hızlı iyileşme için kolayca çevrilebilir anestetik kullanımını içermektedir; trakeotomi içeren olmadan minimal invaziv bir endotrakeal intubasyon; ve göğüs ek insizyon olmadan orijinal torakotomi sitesi aracılığıyla Torasentesiz olduğu gibietkili bir göğüs boşluğundan kan ve havayı çıkarmak için bazı başka yöntemler de yapılır. Bu yöntem nispeten büyük ölçüde, cerrahi ve ameliyat sonrası komplikasyonlar ve ölüm oranını azaltır ve tekrarlanabilirlik geliştirir, diğer yöntemlere göre daha az invaziv.

Fareler yirmi sekiz gün ameliyattan sonra ötenazi uygulandı, ve kalpleri hasat ve incelenmektedir. 75 mg / kg ketamin ve 5 - - 10 mg / kg ksilazin fareler 50 ile anestezi uygulanmıştır. Hayvan yeterli anestezi altında olduğunda, göğüs boşluğu açıldı ve bir 23 gauge iğne kullanarak, soğuk bir potasyum klorid (KCL, 30 mM), kalbin arka taban bölgesine enjekte edilir. Kalp diastolde tutuklandı. ligasyon daha geçerli olabilmesi için, kalp hayvan çıkarılır ve% 4 paraf...

Watch this Scientific Journal Video about Murine Left Anterior Descending LAD Coronary Artery Ligation: An Improved and Simplified Model for Myocardial Infarction at JoVE.com

Murine Left Anterior Descending LAD Coronary Artery Ligation: An Improved and Simplified Model for Myocardial Infarction

We provide a reliable method for left anterior descending artery (LAD) ligation in a mouse model. This method is comparatively less invasive than other methods, involving endotracheal intubation, a left-sided thoracotomy approach, and thoracentesis. This method can be used as a model for both acute and chronic myocardial infarction (MI).

Kemirgen Sol Ön İnen (LAD) koroner arter ligasyonu: Miyokard İnfarktüsü Yönelik Gelişmiş ve Basitleştirilmiş Modeli

Ischemic heart disease (IHD), or acute coronary syndrome (ACS), is one of the leading causes of death in the United States. IHD is characterized by ...

murine-left-anterior-descending-lad-coronary-artery-ligation-an

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

Murine Left Anterior Descending (LAD) Coronary Artery Ligation: An Improved and Simplified Model for Myocardial Infarction

24.4K Görüntüleme.  Rhode Island Hospital, Brown University Warren Alpert Medical School. We provide a reliable method for left anterior descending artery (LAD) ligation in a mouse model. This method is comparatively less invasive than other methods, involving endotracheal intubation, a left-sided thoracotomy approach, and thoracentesis. This method can be used as a model for both acute and chronic myocardial infarction (MI).

Video: Murine Left Anterior Descending LAD Coronary Artery Ligation: An Improved and Simplified Model for Myocardial Infarction

Coronary Artery Ligation and Intramyocardial Injection in a Murine Model of Infarction

Mouse models are a valuable tool for studying acute injury and chronic remodeling of the myocardium in vivo. With the advent of genetic modifications to the whole organism or the myocardium and an array of biological and/or synthetic materials, there is great potential for any combination of these to assuage the extent of acute ischemic injury and impede the onset of heart failure pursuant to myocardial remodeling. 
Here we present the methods and materials used to reliably perform this microsurgery and the modifications involved for temporary (with reperfusion) or permanent coronary artery occlusion studies as well as intramyocardial injections. The effects on the heart that can be seen during the procedure and at the termination of the experiment in addition to histological evaluation will verify efficacy. 
Briefly, surgical preparation involves anesthetizing the mice, removing the fur on the chest, and then disinfecting the surgical area. Intratracheal intubation is achieved by transesophageal illumination using a fiber optic light. The tubing is then connected to a ventilator. An incision made on the chest exposes the pectoral muscles which will be cut to view the ribs. For ischemia/reperfusion studies, a 1 cm piece of PE tubing placed over the heart is used to tie the ligature to so that occlusion/reperfusion can be customized. For intramyocardial injections, a Hamilton syringe with sterile 30gauge beveled needle is used. When the myocardial manipulations are complete, the rib cage, the pectoral muscles, and the skin are closed sequentially. Line block analgesia is effected by 0.25% marcaine in sterile saline which is applied to muscle layer prior to closure of the skin. The mice are given a subcutaneous injection of saline and placed in a warming chamber until they are sternally recumbent. They are then returned to the vivarium and housed under standard conditions until the time of tissue collection. At the time of sacrifice, the mice are anesthetized, the heart is arrested in diastole with KCl or BDM, rinsed with saline, and immersed in fixative. Subsequently, routine procedures for processing, embedding, sectioning, and histological staining are performed. 
Nonsurgical intubation of a mouse and the microsurgical manipulations described make this a technically challenging model to learn and achieve reproducibility. These procedures, combined with the difficulty in performing consistent manipulations of the ligature for timed occlusion(s) and reperfusion or intramyocardial injections, can also affect the survival rate so optimization and consistency are critical.

Numerous genetic manipulations and/or intramyocardial injections of genes, proteins, cells, and/or biomaterials are superimposed upon the dimension of time in studies of acute ischemia/ reperfusion injury and chronic remodeling in mice. This video illustrates the microsurgical procedures for ischemia/reperfusion, permanent coronary artery ligation, and intramyocardial injection studies.

Enfarktüsü bir murin Model Koroner Arter ligasyon ve intramiyokardiyal Enjeksiyon

Mouse models are a valuable tool for studying acute injury and chronic remodeling of the myocardium in vivo.  With the advent of genetic modifications to ...

Tıp

Modified Technique for Coronary Artery Ligation in Mice

Myocardial infarction (MI) is one of the most important causes of mortality in humans1-3. In order to improve morbidity and mortality in patients with MI we need better knowledge about pathophysiology of myocardial ischemia. This knowledge may be valuable to define new therapeutic targets for innovative cardiovascular therapies4. Experimental MI model in mice is an increasingly popular small-animal model in preclinical research in which MI is induced by means of permanent or temporary ligation of left coronary artery (LCA)5. In this video, we describe the step-by-step method of how to induce experimental MI in mice.
The animal is first anesthetized with 2% isoflurane. The unconscious mouse is then intubated and connected to a ventilator for artificial ventilation. The left chest is shaved and 1.5 cm incision along mid-axillary line is made in the skin. The left pectoralis major muscle is bluntly dissociated until the ribs are exposed. The muscle layers are pulled aside and fixed with an eyelid-retractor. After these preparations, left thoracotomy is performed between the third and fourth ribs in order to visualize the anterior surface of the heart and left lung. The proximal segment of LCA artery is then ligated with a 7-0 ethilon suture which typically induces an infarct size ~40% of left ventricle. At the end, the chest is closed and the animals receive postoperative analgesia (Temgesic, 0.3 mg/50 ml, ip). The animals are kept in a warm cage until spontaneous recovery.

The surgical procedure used to induce experimental myocardial infarction in mice begins with left thoracotomy between the third and the fourth ribs in order to visualize the anterior surface of the heart and left lung. The left coronary artery is ligated, the chest is closed and the mouse is allowed to recover spontaneously.

Fare Koroner Arter Ligasyonu için Modifiye Tekniği

Myocardial infarction (MI) is one of the most important causes of mortality in humans1-3. In order to improve morbidity and mortality in patients with MI ...

LAD-Ligation: A Murine Model of Myocardial Infarction

Research models of infarction and myocardial ischemia are essential to investigate the acute and chronic pathobiological and pathophysiological processes in myocardial ischemia and to develop and optimize future treatment.
Two different methods of creating myocardial ischemia are performed in laboratory rodents. The first method is to create cryo infarction, a fast but inaccurate technique, where a cryo-pen is applied on the surface of the heart (1-3). Using this method the scientist can not guarantee that the cryo-scar leads to ischemia, also a vast myocardial injury is created that shows pathophysiological side effects that are not related to myocardial infarction. The second method is the permanent ligation of the left anterior descending artery (LAD). Here the LAD is ligated with one single stitch, forming an ischemia that can be seen almost immediately. By closing the LAD, no further blood flow is permitted in that area, while the surrounding myocardial tissue is nearly not affected. This surgical procedure imitates the pathobiological and pathophysiological aspects occurring in infarction-related myocardial ischemia.
The method introduced in this video demonstrates the surgical procedure of a mouse infarction model by ligating the LAD. This model is convenient for pathobiological and pathophysiological as well as immunobiological studies on cardiac infarction. The shown technique provides high accuracy and correlates well with histological sections.

This video demonstrates how to use a fast and reliable model to study pathobiological and pathophysiological processes of myocardial ischemia.

LAD-ligasyon murin Model: Miyokard Enfarktüsü

Research models of infarction and myocardial ischemia are essential to investigate the acute and chronic pathobiological and pathophysiological processes ...

Biyoloji

A Murine Model of Myocardial Ischemia-reperfusion Injury through Ligation of the Left Anterior Descending Artery

Acute or chronic myocardial infarction (MI) are cardiovascular events resulting in high morbidity and mortality. Establishing the pathological mechanisms at work during MI and developing effective therapeutic approaches requires methodology to reproducibly simulate the clinical incidence and reflect the pathophysiological changes associated with MI. Here, we describe a surgical method to induce MI in mouse models that can be used for short-term ischemia-reperfusion (I/R) injury as well as permanent ligation. The major advantage of this method is to facilitate location of the left anterior descending artery (LAD) to allow for accurate ligation of this artery to induce ischemia in the left ventricle of the mouse heart. Accurate positioning of the ligature on the LAD increases reproducibility of infarct size and thus produces more reliable results. Greater precision in placement of the ligature will improve the standard surgical approaches to simulate MI in mice, thus reducing the number of experimental animals necessary for statistically relevant studies and improving our understanding of the mechanisms producing cardiac dysfunction following MI. This mouse model of MI is also useful for the preclinical testing of treatments targeting myocardial damage following MI.

We introduce a surgical method to induce experimental ischemia/reperfusion (I/R) injury to simulate myocardial infarction (MI) in mouse models that allows for more clarity in positioning of the ligation on the left anterior descending artery (LAD) to increase the reproducibility of MI experiments in mice.

Sol ön inen arter ligasyonu ile Miyokard İskemi-reperfüzyon Hasarı bir fare modeli

Acute or chronic myocardial infarction (MI) are cardiovascular events resulting in high morbidity and mortality. Establishing the pathological mechanisms ...

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.

Farelerde Sol Ön İnen Koroner Arter Daimi ligasyonu: Post-miyokard Enfarktüs Remodelling ve Kalp Yetmezliği Bir Model

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 ...

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.

Sol Anterior İnen Koroner Arterin Kalıcı Ligasyonu ile Murine Miyokard Enfarktüsü Modeli

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

Induction of Myocardial Infarction and Myocardial Ischemia-Reperfusion Injury in Mice

Acute myocardial infarction is a common cardiovascular disease with high mortality. Myocardial reperfusion injury can counteract the beneficial effects of heart reflow and induce secondary myocardial injury. A simple and reproducible model of myocardial infarction and myocardial ischemia-reperfusion injury is a good tool for researchers. Here, a customizable method to create a myocardial infarction (MI) model and MIRI by precision ligation of the left anterior descending coronary artery (LAD) through micromanipulation is described. Accurate and reproducible ligature positioning of the LAD helps obtain consistent results for heart injury. ST-segment changes can help to identify model accuracy. The serum level of cardiac troponin T (cTnT) is used to assess the myocardial injury, cardiac ultrasound is employed to evaluate the myocardial systolic function, and Evans-Blue/triphenyl tetrazolium chloride staining is used to measure infarct size. In general, this protocol reduces procedure duration, ensures controllable infarct size, and improves mouse survival.

Here we describe a simple and reproducible method that can induce myocardial infarction or myocardial ischemia-reperfusion injury in mice by precision ligation of the left anterior descending coronary artery through micromanipulation.

Farelerde Miyokard Enfarktüsü ve Miyokard İskemi-Reperfüzyon Hasarının İndüksiyonu

Acute myocardial infarction is a common cardiovascular disease with high mortality. Myocardial reperfusion injury can counteract the beneficial effects of ...

Left Coronary Artery Ligation: A Surgical Murine Model of Myocardial Infarction

Ischemic heart disease and subsequent myocardial infarction (MI) is one of the leading causes of mortality in the United States and around the world. In order to explore the pathophysiological changes after myocardial infarction and design future treatments, research models of MI are required. Permanent ligation of the left coronary artery (LCA) in mice is a popular model to investigate cardiac function and ventricular remodeling post MI. Here we describe a less invasive, reliable, and reproducible surgical murine MI model by permanent ligation of the LCA. Our surgical model comprises of an easily reversible general anesthesia, endotracheal intubation that does not require a tracheotomy, and a thoracotomy. Electrocardiography and troponin measurement should be performed to ensure MI. Echocardiography at day 28 after MI will discern heart function and heart failure parameters. The degree of cardiac fibrosis can be evaluated by Masson's trichrome staining and cardiac MRI. This MI model is useful for studying the pathophysiological and immunological alterations after MI.

Presented here is a surgical procedure for permanent ligation of the left coronary artery in mice. This model can be used to investigate the pathophysiology and associated inflammatory response after myocardial infarction.

Sol Koroner Arter Ligasyonu: Miyokard Enfarktüsünün Cerrahi Murin Modeli

Ischemic heart disease and subsequent myocardial infarction (MI) is one of the leading causes of mortality in the United States and around the world. In ...

Left Anterior Descending Coronary Artery Ligation for Ischemia-Reperfusion Research: Model Improvement via Technical Modifications and Quality Control

Coronary heart disease is the leading cause of death globally. Complete cessation of blood flow in coronary arteries causes ST-segment elevation myocardial infarction (STEMI), resulting in cardiogenic shock and fatal arrhythmia, which are associated with high mortality. Primary coronary intervention (PCI) for recanalizing the coronary artery significantly improves the outcomes of STEMI, but advancements made in shortening the door-to-balloon time have failed to reduce in-hospital mortality, suggesting that additional therapeutic strategies are required. Left anterior descending coronary artery (LAD) ligation in rats is an animal model for acute myocardial IR research that is comparable to the clinical scenario in which rapid coronary recanalization through PCI is used for STEMI; however, PCI-induced STEMI is a technically challenging and complicated operation associated with high mortality and great variation in infarction size. We identified the ideal position for LAD ligation, created a gadget to control a snare loop, and supported a modified surgical maneuver, thereby reducing tissue damage, to establish a reliable and reproducible acute myocardial ischemia-reperfusion (IR) research protocol for rats. This is a non-survival surgery. We also propose a method for validating the quality of study results, which is a critical step for determining the accuracy of subsequent biochemical analyses.

Herein, we present a protocol focusing on the quality control of the left anterior descending coronary artery ligation by technically modifying the traditional procedure in rats for acute myocardial ischemia-reperfusion research.

İskemi-Reperfüzyon Araştırmaları için Sol Ön İnen Koroner Arter Ligasyonu: Teknik Modifikasyonlar ve Kalite Kontrol Yoluyla Model İyileştirme

Coronary heart disease is the leading cause of death globally. Complete cessation of blood flow in coronary arteries causes ST-segment elevation ...

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.

<meta charset="utf8"></head><body>Nakavt farelerde kardiyak onarım ve yeniden şekillenmeyi incelemek için deneysel bir miyokard enfarktüsü modeli

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.

Nakavt farelerde kardiyak onarım ve yeniden şekillenmeyi incelemek için deneysel bir miyokard enfarktüsü modeli

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