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.

이 동물 프로토콜을 검토하고로드 아일랜드 병원의 기관 동물 관리 및 사용위원회 (IACUC)에 의해 승인되었습니다. 1. 마취 및 삽관 <ol><li> 수술 후 진통제의 복용량을 계산하기 위해 마우스의 무게를. </li><li> 유도 챔버에 마우스를 넣고 9 4 % 이소 플루 란을 전달 - 걸쳐 동물 모니터링 10 분. 상기 장치는 약 250 ℃로 예열 할 수 있도록 핫 비드 살균기 켜기. 20 분 - 예열은 15 일이 소요됩니다. </li><li> 마우스 약 32 호흡 / 분의 호흡 속도 마취 깊이 평면에 도달하면, 스티로폼 보드에 마우스 앙와위 배치 오픈 입을 보유하는 상부 앞니에 따라 고정하는 탄성 밴드를 사용한다. 발가락 핀치를 수행하여 진정 작용을 확인합니다. 구강 인두가 가시화 될 수 있도록 마우스보다 높은 강도 조명을 배치합니다. </li><li> 턱과 다른 쌍을 열 곡선 집게를 사용하여집게의 길 밖으로 혀를 들어 올립니다. 또는 약간 마우스의 몸 눈높이 아래에 위치하면서 삽관해야합니다. 수술 돋보기를 사용하는 것이 좋습니다. </li><li> 성대의 개폐를 시각화합니다. 개방되면, 삽입 20 게이지, 1-에 무딘 팁 니들 도입기와 정맥 카테터. 기관 개방 도관을 안내하지만, 기관에 바늘을 삽입 방지하기 위해 바늘을 사용한다. 정확한 위치의 확인은 플라스틱 전송 피펫을 사용하여 수행 할 수 있습니다. </li><li> 가열 장치를 구비 한 조작면에 삽관 마우스 이동. 150 μL / 스트로크의 행정 용적 / 분 130 스트로크의 스트로크 속도 설정 작은 설치류 호흡기 마우스를 연결한다. </li><li> 2.5 %의 이소 플루 란을 제공합니다. 양자 가슴 상승에 대한 확인하여 삽관을 확인합니다. 발가락 핀치를 수행하여 마취를 확인합니다. 완전히 마취되기 위해 인공 호흡기에 10 분 - 마우스는 5 필요할 수 있습니다. </li></ol><ol><li> 인공 호흡기와 IV 카테터 사이의 연결 부위에서 삽관 튜브 아래 테이프. 사지 아래 테이프. 눈에 멸균 윤활 방울을 놓습니다. </li><li> 전기 면도기와 흉부의 복부 왼쪽 트림. 건조 와이프와 면도 모피를 먼지와 멸균 면봉을 사용하여 제모 크림의 작은 레이어를 적용합니다. 45 초 - 크림은 약 30 모낭과 접촉을 유지해야한다. </li><li> 크림 프로세스 동안 담가 Betadine 가득 세 1.5mL 용량 튜브에 세 멸균 면봉을 놓는다. 증류수에 적신 헝겊을 사용하여 부드럽게 크림 및 모피을 닦아. </li><li> 주변에 중심에서 이동하는 원 운동의 청소, Betadine 멸균 70 % 이소프로판올 준비 패드 교번 수술 부위 세번 세척. 마우스의 수술 필드 위에 분기 크기의 구멍이있는 멸균 드레이프를 놓습니다. </li><li> MOUS 주변 지역을 청소70 % 에탄올로 전자. 한 번 더 발가락 핀치와 마취를 확인합니다. </li></ol> 3. LAD 내고 <ol><li> 대략 20 초 동안 250 ℃로 예열 된 핫 비드 멸균기에서 멸균 수술 도구를 놓는다. 멸균 멸균 된 수술 용 드레이프에 살균 장비를 놓습니다. 돈 수술 장갑. </li><li> 부드럽게 저명한 칼 모양의 연골의 왼쪽 지점에서 약 5 mm의 피부를 들어 올려 좋은 팁 집게를 사용합니다. 흉골의 수준으로, 위쪽이 시점에서 피부에 수직 절개를 만들기 위해 10 번 블레이드와 수술 메스를 사용합니다. </li><li> 부드럽게 피부와 근육 층을 분리 곡선 집게를 사용합니다. 피부 절개 다음, 근육 층을 엽니 다. 근육 층, 절개의 양쪽에 하나를 통해이 5-0 폴리 프로필렌 봉합사를 삽입하고 근육 층이 열려 개최 클램프 일시적으로 봉합을 고정합니다. </li><li> 다음, 확인하고 세 번째 늑간 공간에서 절개를 만드는흉곽의 자연 각도. 마우스의 왼쪽 사지에서 테이프를 제거하고 테이프로 오른쪽 뒷발에 자사의 뒷면 왼쪽 발을 고정합니다. 테이프의 긴 조각을 잘라 약간 상승 된 위치에 운영 표면의 왼쪽 앞발을 고정합니다. 70 % 에탄올로 장갑을 청소합니다. </li><li> 부드럽게 3 번째와 4 번째 갈비뼈를 벌리 견인기를 사용합니다. 약 1 × ½에있어서, 멸균 거즈의 작은 부분을 절단하고, 0.9 % 멸균 식염수에 담근다. 여분의 염분을 꽉 잡고 조심스럽게 절차를 수행하는 동안 실수로 폐 손상을 방지하기 위해 왼쪽 폐에 거즈를 삽입하는 집게를 사용합니다. </li><li> 부드럽게 집게로 얇은 심낭을 제거합니다. </li><li> 멸균 면봉 떨어져면 소량의 눈물과 작은 공로를 굴립니다. 멸균 0.9 % 식염수에이 목화 공을 찍어 부드럽게 동맥을 감사하는 마음의 표면에 면봉. 부드럽게 위쪽 왼쪽 귓바퀴를 밀어에서 관상 동맥를 찾습니다일 Neath. </li><li> LAD를 식별하고 LAD 하에서 8-0 나일론 봉합사를 전달; 이 완전한은 결찰을 확보하기 위해 발생합니다. 결찰에 성공하면 합자 의지 희게에서 좌심실의 원위부. </li><li> 집게를 사용하여 이전에 삽입 된 거즈를 제거하고 부드럽게 견인기를 제거합니다. 개흉술을 통해 개구 흉강으로 25 게이지 바늘에 부착 된 6에서 25 게이지 플렉시블 튜브를 삽입한다. 왼쪽 폐 위의 공간에 튜브의 2 - 약 1 사전. 부정사 위치에 마우스를 반환하고 70 % 에탄올로 장갑을 청소합니다. </li><li> 장소에 가슴 튜브를 유지, 흉곽을 닫습니다 간단한 중단 패턴에서 5-0 폴리 프로필렌 봉합사를 사용합니다. 열린 근육 층을 고정하는 두 개의 봉합사를 제거합니다. 다시 자리에 가슴 튜브를 유지, 근육 층을 닫습니다 간단한 연속 패턴에서 5-0 폴리 프로필렌 봉합사를 사용합니다. </li><li> 흉관의 25 게이지 바늘로 1 ml 주사기를 첨부. 부드럽게 런지에 위쪽으로 당겨R 동시에 서서히 집게 흉강에서 흉관 추출있다. 이 단계에서 달리 기흉 흉강 및 결과에 포획 될 것이다 과잉 공기와 혈액을 제거로 천천히 튜브를 추출한다. </li><li> 주사기가 가득 차면, 바늘에서 주사기를 분리하고 폐기물 비커 또는 싱크에 폐기물 폐기하십시오. 가슴 튜브가 완전히 추출 될 때까지이 과정을 계속합니다. 가슴이 밀봉되어 있는지 확인합니다. </li><li> 1.5 %로 이소 플루 란을 줄이십시오. 간단한 중단 패턴에서 4-0 폴리 프로필렌 봉합사로 피부를 닫습니다. 오프 이소 플루 란 기화기를 돌립니다. </li><li> 복강 내 (IP) 주입을 통해 0.9 % 식염수 중 0.1 ㎎ / ㎖의 부 프레 노르 핀을 투여. 국소 절개 0.9 % 식염수에 2 ㎎ / ㎖ 부피 바카 2 ㎎ / ㎖ 리도카인을 적용한다. 마우스의 체중을 식염수 양을 스케일링, 피하 주사를 통하여 0.9 % 식염수 500 μL - 200 사이를 관리. </li><li> 된 관 후 5 분을 기다립니다통증 약물을 stering은 삽관 튜브에서 마우스를 제거합니다. 이 인공 호흡기 오프 전환에 도움이됩니다. <ol><li> 마우스가 인공 호흡기 떨어져 한 번 양자 가슴 상승이없는 경우, 바늘 압축 해제를 수행합니다. 이 저항의 급격한 감소에 의해 표시 흉강을 입력 할 때까지이를 위해, 25 게이지 바늘 및 멸균 3 번째 및 4 번째의 리브 사이를 1ml 주사기를 소개한다. 과잉 공기를 제거하기 위해 플런저를 부드럽게 잡아 당깁니다. </li></ol></li><li> 마우스가 적절한 양자 간 호흡 속도와 깊이를 보여주고 발가락 핀치에 응답 할 때, 가열 램프 아래에 깨끗한 복구 케이지에서 마우스를 놓습니다. 20 분 - 15 층류 후드에서 모니터링, 습기 음식과 물 병 마우스를 제공합니다. 과장된 호흡 노력, 과도한 출혈, 또는 기타 잠재적으로 생명을 위협하는 합병증에 대한 모니터링합니다. </li><li> 앞으로 3 일 들어, IP를 통해에서 0.1 ㎎ / ㎖ 프레 노르 핀 진통제를 투여하루에 두 번 배제 력. 매일 마우스를 모니터합니다. </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.

실험실에서 MI 모델의 사용이 증가, 설명 된 절차는 수술 후 통증과 불편을 최소화하면서 마우스의 효율성과 생존율을 높이기 위해 노력한다. 이 프로토콜은 LAD의 결찰 과정의 다양한 측면에 많은 개선함으로써 사망률을 최소화하기 위해 노력하고 있습니다. 몇 가지 차이점이 있습니다. 때문에 마취의 그 이상 기간의 이익을 위해, 유도 이소 플루 란과 함께 케타민과 자일 라진을 활?...

관상 동맥 질환, 또는 ACS, 가장 널리 심혈관 이벤트 2020 년 1 이환율과 전세계 사망의 주요 원인으로 간주됩니다. ACS의 원인으로 인해 심장 조직이 혈액 흐름을 감소 블록 또는 관상 동맥 경화성 플라크의 파열에 심근 혈전증의 존재이다. 따라서, 예컨대 심근 경색 (MI) 3, 4, 급성 심근 허혈의 존재와 일치하는 임상 적 징후가있다. MI는 심실 기능 부전과 심부전 5, 6으로 이어질 수있는 심근의 질량 손실과 병적 인 심실 리모델링에 진행에 연결됩니다. IHD을 연구하는 가장 효과적인 방법 중 하나는 동물 모델에서 인간의 심근 경색을 모방하는 것이 었습니다. 이것은 LAD의 결찰에 의해 달성된다마우스. 이 모델을 사용하여, 우리는 마음이 허혈성 심질환으로 인한 손상으로부터 보호 할 수있는 방법을 연구한다. 지난 10 년간, 연구자들은 쥐에 쥐의 변화를 포함하여, 작은 동물에 더 큰 동물 모델을 사용하는 이동했다. 작은 마우스 모델은 작은 크기를 포함하여 많은 이유, 대형 쓰레기의 크기, 유지하기 위해 저렴한 비용, 짧은 임신 기간뿐만 아니라 형질 전환 유전자 녹아웃 모델 (7)의 넓은 가용성에 대한 선호되기 시작한다. 마우스는 작은 규모이지만, 특별히 그들을 위해 디자인 된 새로운 수술 도구이 개발에 도움을했다. 우리의 방법은 새로운 수술 도구를 사용합니다. 여러 가지 방법이 침입 기관 절개술을 구현하는 동안, 우리는 기관 내 삽관의 덜 침습적 인 방법을 사용합니다. 구강 인두의 오버 헤드 조명을 사용하여, 우리는 t을위한 안전하고 덜 충격적인 경험을 제공, 어떤 절개를 만들지 않고 삽관그는 동물. 마우스는 인공 호흡기에 배치하고, 전체 과정 동안 이소 플루 란에 보관됩니다. 이 중단되면 동물이 마취에서 회복하기 때문에 약물에 의해 생산 마취의 짧은 기간에, 그것은 단지 몇 분 정도 걸립니다. 우리의 외과 모델은 또한 최소 침습 천자가 포함되어 있습니다. 인장 기흉 : 일본어 개흉술을 통해 천자를 사용 흉강 혈액 및 과잉 공기의 제거는 조심 LAD 결찰하는 일반적인 수술 후 합병증을 해결했다. 다른 방법 - 하나 천자-했다 굴복 적은 수술 합병증을위한 기관 절개술에 대한 다른에 사용되는 두 개의 추가 절개에 대한 필요성을 제거하고 크게 사망률을 감소시켰다이 방법.

<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

허혈성 심장 질환 (IHD), 급성 관상 동맥 증후군 (ACS)는 미국에서 사망의 주요 원인 중 하나입니다. IHD는 산소의 감소 및 심장 근육의 괴사 계속되는 결과 심장에 혈액 공급이 감소 특징으로한다. MI 모델은 단기 허혈 - 재관류 모델과 장기 영구 결찰 모델로의 사용에 대한 인기를 얻고있다. 아래, 우리는 LAD의 영구 결찰을위한 신뢰할 수있는 방법을 설명합니다. 마우스 유전 공학 기술이 더 진보되고, 품질 쥐의 수술 도구의 증가 가용성으로, 마우스는 MI 수술에 대한 인기 모델이되고있다. 우리의 외과 모델 마우스의 빠른 복구를 위해 쉽게 되돌릴 수 마취제의 사용을 포함; 기관 절개술을 포함하지 않고 최소 침습 기관 내 삽관; 상기 가슴 절개 부가 만들지 않고 원래 개흉술 사이트를 통해 천자는 그대로효과적으로 흉강의 과도한 혈액과 공기를 제거하고, 일부 다른 방법으로 수행. 이 방법은 비교적 급격히 외과 수술 합병증 및 사망률 감소 및 재현성을 개선하는 다른 방법에 비해 덜 침습적이다.

생쥐 이십팔일 수술 후 안락사되고, 마음은 수확을 조사하고 있습니다. 75 ㎎ / ㎏의 케타민 및 5 - - 10 밀리그램 / kg 자일 라진 마우스에게 50 마취. 동물이 충분한 마취되면, 흉강을 개방하고, 23 게이지 바늘을 사용하고, 차가운 염화칼륨 (KCL, 30 mM)을 심장의 후부 기저 영역으로 주입된다. 심장은 이완기에서 체포된다. 결찰의 추가 검증을 위해, 마음은 동물에서 제거되고 4 %...

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).

쥐 왼쪽 앞쪽에 내림차순 (LAD) 관상 동맥 결찰 : 심근 경색에 대한 개선 및 단순화 된 모델

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 조회수.  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).

비디오: 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.

경색의 Murine 모델에서 관상 동맥 내고과 Intramyocardial 사출

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 ...

연구

의학

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.

마우스의 관상 동맥 결합에 대한 수정 기술

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.

내사 - 결합 : 심근 경색의 Murine 모델

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

생물학

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.

왼쪽 앞쪽에 내림차순 동맥의 결찰을 통해 심근 허혈 - 재관류 손상의 생쥐 모델

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.

마우스의 왼쪽 전방 내림차순 관상 동맥의 영구 결찰 : 심근 경색 리모델링과 심장 마비의 모델

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.

왼쪽 전방 내림차순 관상 동맥의 영구 결찰을 사용하는 뮤린 심근 경색 모델

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.

마우스의 심근경색 및 심근 허혈-재관류 손상 유도

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.

좌측 관상동맥 결찰술: 심근경색의 외과적 쥐 모델

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.

허혈 재관류 연구를 위한 좌전하행 관상동맥 결찰술: 기술적 수정 및 품질 관리를 통한 모델 개선

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.

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

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