이 작품은 스위스 국립 과학 재단 부여 150271에 의해 지원 되었다.

이 실험 프로토콜 Cantonal 수의 사무실, Fribourg는 스위스 연방 수의 사무실, 스위스 (FR 2015/58)에 의해 승인 되었습니다.
 참고: 남성 NZW 토끼 2.8 3.2 k g 사이 무게 사용 되었다. 동물 (12 h 빛 및 어두운 주기, ad libitum 물과 식량 제공) 기존의 조건 하에서 보관 되어 있었다. 풍선 노출 전에 동물 1 주 동안 그들은 정상적인 차 다이어트 지 루 했다에 대 한 acclimated 했다. 순응의 1 주 후 토끼 높은 지방 (8.6%)의 구성 하는 atherogenic 다이어트에 전환 했다 하 고 포화 지방산 205 mg/kg (1%) 콜레스테롤 다이어트 전체 연구 기간에 대 한. 왼쪽 장 골 동맥에서 풍선 손상 다이어트 개시 후 1 주 수행 및 동물 풍선 부상 4 주 또는 2 주 후 희생 되었다.
 1. 수술 전 절차 <ol><li>유리 구슬 소독 기 또는 다른 적합 한 악기와 함께 사용 하기 전에 모든 수술 기구를 소독.</li>
	<li>준비 풍선 카 테 터 어셈블리를 확인 하 고.
	<ol><li>첨부 1 mL luer 잠금 주사기 luer 잠금 부분 풍선 카 테 터의 정상적인 염 분으로 가득합니다. 신중 하 게 덫을 놓은 공기의 부재에 대 한 모니터링 합니다. 누수를 확인 하 고 주사기의 플런저를 눌러 적절 한 풍선 인플레이션 확인.</li>
	</ol></li> <li>토끼의 무게 및 37 thermopad 설정 ° c.</li>
	<li>Buprenorphine 솔루션을 사용 하 여 0.3 밀리 그램의 농도에 / ml. 0.01 mg/kg의 복용량을 피하 주사 하.</li>
	<li>Anesthetize 10-15 분에 대 한 유도 실에서 5 L/분 O 2와 5 %isoflurane 토끼</li>
	<li>외과 플랫폼에 계속 생리대에 마 취 토끼를 놓습니다. 패치 및 온도, 호흡, 심전도 모니터링 하는 클립.</li>
	<li>적합 한 마 취 기계에 연결 된 얼굴 마스크를 토끼의 주 둥이 연결 합니다. 마 취와 isoflurane 2.5 L/min O 2 (4.0%)를 유지 합니다. 적절 한 anesthetization (개 그, pinnae 반사의 손실 및 근육의 부족으로 표시)를 확인 하십시오.</li>
	<li>안과 연 고는 각 막 건조 하지 않도록 두 눈에 적용 됩니다. 노출만 더 낮은 사지와 토끼를 메 마른 외과 시트와 함께 드러 워 진.</li>
</ol> 2. 수술 프로토콜 <ol><li>동물 발기를 사용 하 여 무릎 관절 바로 아래 복 부 영역에서 제거.</li>
	<li>적당 한 살 균 제는 피부를 청소 하 고 느슨한 머리를 제거 하는 영역을 면봉.</li>
	<li>Saphenous 동맥을 찾아서 메스를 사용 하 여 길이에 약 1.5 ㎝의 작은 피부 절 개를 하 게.</li>
	<li>대 퇴 정 맥, 대 퇴 신경 손상 없이 작은 곡선된 겸 자도 saphenous 동맥의 작은 부분을 노출.</li>
	<li>Saphenous 동맥 아래 두 개의 느슨한 합자 루프 (5-0 실크) 놓고 한 합자 루프는 동맥의 말 초 끝을 묶어. 장 골 동맥에서 혈액의 흐름을 막으려고 합자 위에 microvascular 클램프 장소.</li>
	<li>Papaverine 동맥 팽창 하 고 vasospasm을 방지 하기 위해 한 방울 몸에 붙이기도 적용.</li>
	<li>묶인된 합자의 도움으로 saphenous 동맥을 올리고 확인 작은 arteriotomy 절 개는 24를 사용 하 여 게이지 바늘.</li>
	<li>미세 집게와 절 개 플랩 상승 하 고 천천히 동맥의 루멘으로 정 맥 선택 또는 지도 바늘을 삽입.</li>
	<li>Saphenous 동맥에 2 프랑스어 Fogarty 동맥 embolectomy 카 테 터를 삽입합니다. 정 맥 선택 및 microvascular 클램프 제거.</li>
	<li>6 마크 (20-25 cm)에 해당 하는 위치 약까지 카 테 터를 미리 장 골 분기점 위에 2-5 cm.</li>
	<li>1 mL 주사기를 사용 하 여 0.1 mL 정상적인 염 분 또는 6의 공칭 압력에서 풍선을 부 풀 려 16 , 22에 설명 된 대로 규제 수동 공기 펌프를 사용 하 여 atm.</li>
	<li>집게와 풍선 카 테 터를 누른 테 회전 하는 동안 삽입 지점 장 골 arterytoward 통해 6 cm에 의해 다시 끕니다.</li>
	<li>주사기의 플런저를 후퇴 하 여 풍선을 수축.</li>
	<li>완료 내 피 노출 되도록 단계 2.10 2.13 세 번 반복.</li>
	<li>카 테 터를 제거 하 고 출혈을 멈추게 하는 arteriotomy 사이트의 바로 위에 합자 루프를 즉시 넥타이.</li>
	<li>상처 주위 적합 한 살 균 모두를 적용 하 고 혈 병을 떨어져 면봉. 5-0 봉합, 피부 절 개를 닫고 povidone-요오드 솔루션 수술 사이트 소독.</li>
	<li>2.1에 2.16는 contralateral에 장 골 새로운 카 테 터를 사용 하 여 단계를 반복 하십시오.</li>
	<li>눈에서 안과 연 고를 면봉.</li>
</ol> 3. 수술 치료 <ol><li>sulfadoxine 관리 40 mg/kg 및 trimethoprim 8 mg/kg 또는 다른 적합 한 항생제는 수술 직후.</li>
	<li>마 취 회복 기간 동안 깨끗 한 압력가 감 금 소에 열 패드를 통해 토끼를 유지.</li>
	<li>모니터링 패치 및 클립 제거.</li>
	<li>복구 후 자신의 집 새를 토끼를 반환. 주사 피하 buprenorphine 0.05-0.1 mg/kg 게시물-operatively 48 헤 또 다른 2 주 또는 4 주에 대 한 계속 atherogenic 다이어트에 대 한 모든 6-12 h.</li>
</ol> 4. 조직 수확 및 분석의 패 구성 <ol><li>2 주 (초기 얇은 패)에 대 한 풍선 상해의 3 주 후, 위에서 설명한 대로 isoflurane 비슷한 방식으로 사용 하 여 토끼 anesthetize.</li>
	<li>흉 강 열고 intracardial exsanguination에 의해 토끼를 안락사.</li>
	<li> 23에 설명 된 대로 장 골 동맥을 분리.
	<ol><li>짧게, 복 부를 열고는 retroperitoneum 노출. 장 골 분기점 쪽으로 대동맥을 추적 하 고 분기점 위에 그것을 넥타이. 조심 스럽게 노출 하 고 격리 두 장 골 동맥 주위 조직을 제거.</li>
	</ol></li> <li>Dissect 두 장 골 동맥과 버퍼링 차가운 인산 염에 그들을 담가. 집게의 도움으로 혈전을 제거 합니다. 각 장 골 동맥 characterise 걸쳐 동맥 플 라크의 두께를 4-6 세그먼트로 분할.</li>
	<li>즉시 형 최적의 절삭 온도 화합물, 포함 된 스냅-동결 액체 질소를 사용 하 여 동맥 세그먼트를 포함 하 고-70 ° C. 준비 5 µ m 24에 설명 된 대로 cryostat를 사용 하 여 두꺼운 섹션에서 그것을 유지.</li>
	<li>수행 조직학, 면역 형광 검사 또는 10 , 25에 설명 된 대로 morphometry, 플 라크 지질 및 세포질 내용에 대 한 얼룩 immunohistochemical. 
	참고: 짧게, 버퍼링 하는 인산 염 (PBS)와 동맥 섹션 린스 하 고 0.2%를 사용 하 여 permeabilize 트리톤. PBS 가진 섹션 린스를 안티 α-SM 걸 (1: 200) 또는 RAM11 항 체 (1: 200) 37 ° C에서 1 시간에 대 한 섹션 2% 소 혈 청 알 부 민 30 분 품에 대 한 비 특정 사이트를 차단 합니다. PBS 가진 섹션 린스 하 고 PBS 37 ° C. 씻어 다시 30 분에 대 한 적절 한 이차 항 체와 그들을 품 어 Hoechst 추가 (5 µ g/mL) 10 분 핵 감지.</li>
</ol>

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A., Mosse, P. R., Safstrom, J. A., Young, S. B., Campbell, G. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Balloon+catheter+injury+to+rabbit+carotid+artery.+I.+Changes+in+smooth+muscle+phenotype.">Balloon catheter injury to rabbit carotid artery. I. Changes in smooth muscle phenotype.</a> Arteriosclerosis. 9, 289-298 (1989).</li><li>Miyake, 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=Prevention+of+neointimal+formation+after+angioplasty+using+nuclear+factor-kappaB+decoy+oligodeoxynucleotide-coated+balloon+catheter+in+rabbit+model.">Prevention of neointimal formation after angioplasty using nuclear factor-kappaB decoy oligodeoxynucleotide-coated balloon catheter in rabbit model.</a> Circ Cardiovasc Interv. 7, 787-796 (2014).</li><li>Fulcher, J., Patel, S., Nicholls, S. 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저자 아무 경쟁 금융 관심사를 선언합니다.

토끼 장 골 동맥 경화 모델은 아 테 롬 연구에서 널리 이용 된다. 이 프로토콜은 토끼 빠르게 자발적인 병 변만 콜레스테롤 다이어트와 함께 개발에 비해 더 심각 하 고 고급 패 개발. 중요 한 것은, 동물 수술에서 신속 하 게 복구합니다.
Atherogenesis 위한 주요 자극은 endothelium 부상 하 고 혈관 벽26distends 풍선 카 테 터로 인 한 기계적 손상 이다. 이 이렇게 대 식...

취약 한 동맥 경 화성 플 라크의 파열1선진국 사망의 주요 원인 중 하나입니다. 지난 수 십년 동안 연구 패 진행에 관련 된 여러 분자 및 세포 메커니즘을 펼쳐는, 뿐만 아니라 질병 진행의 복잡 한 메커니즘을 해명 뿐만 아니라 새로운 테스트 노력 필요 아직도 계속 치료 접근 한다. 여러 동물 모델 연구는 동맥 경화를 제안 되었습니다. 유전자 조작, 콜레스테롤 먹이 또는 기계적 피 상해는 쥐, 토끼 또는 minipigs를 포함 하 여 동맥 경화의 가장 동물 모델에서 공유 하는 표준 전략. 이러한 가운데, NZW 토끼에 민감합니다 콜레스테롤 다이어트 동안 정상적인 쥐와 생쥐 마음이 크게 식이 콜레스테롤2,,34를 흡수 하지 않습니다. 토끼는 저절로 대동맥 병 변 세포 콜레스테롤 풍부한 다이어트5,6먹 일 때 일부 섬유 구성 요소와 함께 풍부한 개발. 그러나, 동맥 경 화성 plaquesby 콜레스테롤 다이어트 혼자6,7 먹이 유도 하기 위해 4-8 개월의 긴 준비 시간 실험 설정의 대부분을 위한 주요 결점은. 상대적으로 짧은 시간에 병 변을 유발에 대 한 추구, 높은 콜레스테롤 다이어트와 풍선 상해의 조합 Baumgarter 및 Studer8에 의해 개발 되었습니다. 이 기술은의 전반적인 목표는 2 주 이내에 콜레스테롤 토끼에 거품 세포 (인간에서 지방 조 흔과 유사)의 구성 된 동맥 경 화성 플 라크를 유도 하는 것입니다. 현재 기술 NZW 노인 토끼의 장 골 동맥으로 고급 풍선 카 테 터를 사용 하 여 Baumgarter의 방법에 따라 동맥 벽 상해의 절차를 설명 합니다.
콜레스테롤 풍부한 음식과 함께 부상 풍선 유도 드 endothelialization에서 동맥 경화 이어질 것입니다. 풍선 부상 동맥 경 화성 병 변 형성을 촉진 하 고 균일 한 크기와 분포의 플 라크를 생성 합니다. 내 두껍게 상해 다음 몇 일 이내에 시간과 내 세포 침투 시작의 기간 동안 증가 합니다. 상당한 대 식 세포와 지방 줄무늬 풍선 상해의 7-10 일 후에 나타나기 시작 하 고 미국 심장 협회에서 분류에 따라 병 변의 유형 II로 표시 됩니다. 토끼 풍선 부상은 종종 플 라크 구성 공부 대동맥에서 수행 됩니다. Neointimal 내 피 세포 접착 분자의 높은 수준의 표현. 플 라크 중간 절 개 및 adventitial 변화에 연관 됩니다. 동맥 경 화성 병 변 지질, 확산 부드러운 근육 세포 (SMCs), 교원 질 섬유 및 재생성된 endothelium 아래에 축적 하 고 자연에서 주로 유형 II는 염증 세포의 구성 됩니다. 토끼 플 라크의 토폴로지 배포 원칙에 인간 aortas 9,10 에 보고 유사 했다, 대동맥 장 골 동맥에 비해 크기가 큰 큰 길이에서 플 라크를 생산할 것 이라고. 그러나, 토끼에 있는 아 테 롬의 사이트로 장 골 동맥을 사용 하 여의 주요 장점은 접근, 인간 관상 동맥11, 균일 한 병 변 개발12, 높은 조직 요소 근육 콘텐츠에서 유사성은 활동13 그리고 형태학 및 angiographic 끝점을 상업적으로 제조 된 소자의 평가 허용 하는 인간의 관상 동맥에 비해 일관 된 배 차원. 침략과 비-침략 적 방법 살아있는 동물에서 토끼 장 골 동맥에 플 라크를 분석 조사 있다. 이전 보고서 자기 공명 또한 2.35 테슬라 미스터 시스템 14 의 도움으로 (MRI를) 영상 사용 설명, 혈관 내 초음파 (IVUS) 또는 광학 일관성 단층 촬영 (OCT) 카 적당 수 이미지에 적용 토끼 장 골 동맥에 동맥 경 화성 플 라크입니다. 장 골 동맥은 초음파 이미징 고해상도 진단법을 사용 하는 경우 그리고 대동맥이 기술 또한 탐험 수 있습니다.
지난 10 년간에서이 토끼 모델 풍선 부상의 더욱 패 진행15, 플 라크 회귀16의 메커니즘을 이해 하 있었습니다. 새로운 치료제 스타 틴, 표준 antiplatelet 에이전트, 항 산화 제17,18 everolimus 같은 약물 방출 스 텐트 등의 영향을 연구 하는 모델 사용 되었습니다 또한, 또는 zotarolimus 방출 스텐19,20 neointimal 두껍게. 이 모델은 또한 혈관 내 영상 카 테 터21이미징 근 적외선 형광의 조사에 사용 되었습니다.

<table><tbody><tr><td>New Zealand White rabbits</td><td>Charles River laboratories,France</td><td>Cre:KBL(NZW)</td><td></td></tr><tr><td>Cholesterol rich diet</td><td>Ssniff spezialdi&amp;auml;ten</td><td>Ssniff EF K High Fat and Cholesterol</td><td></td></tr><tr><td>Glass bead sterilizer-Germinator 500</td><td>VWR, Leicestershire, UK</td><td>101326-488</td><td></td></tr><tr><td>Fogarty balloon embolectomy catheters, 2 French</td><td>Edwards Lifesciences, Switzerland</td><td>120602F</td><td>For single use only</td></tr><tr><td>Luer Lock Syringe</td><td>Becton, Dickinson and Company, USA</td><td>309628</td><td></td></tr><tr><td>Thermopad Type 226</td><td>Solis, Switzerland AG</td><td>397387</td><td></td></tr><tr><td>Buprenorphine- Temgesic</td><td>Reckitt Benckiser AG, Switzerland</td><td>7.68042E+12</td><td></td></tr><tr><td>Isoflurane</td><td>Piramal Critical Care, Inc, Bethlehem, PA 18017</td><td>2667-46-7</td><td></td></tr><tr><td>Anaesthesia machine-combi-vet Base Anesthesia System</td><td>Rothacher Medical GmbH, Switzerland</td><td>CV 30-301-A</td><td></td></tr><tr><td>Cardell touch veterinary vital signs monitor</td><td>Midmark, Ohio, USA</td><td>8013-001</td><td></td></tr><tr><td>Ophthalmic ointment-Humigel</td><td>Virbac, France</td><td></td><td></td></tr><tr><td>Animal hair clippers</td><td>Aesculap AG, Germany</td><td>GT420</td><td></td></tr><tr><td>Disinfectant-Betadine solution</td><td>MundipharmaMedicalCompany, Switzerland</td><td>14671-1203</td><td></td></tr><tr><td>Dumont #7 Forceps</td><td>FST Germany</td><td>11274-20</td><td></td></tr><tr><td>Medium and small microscissors</td><td>Medline International Switzerland S&amp;agrave;rl</td><td>UC4337</td><td></td></tr><tr><td>Microvascular clamps</td><td>FST, Germany</td><td>18051-28</td><td></td></tr><tr><td>Papaverine</td><td>ESCA chemicals, Switzerland</td><td>RE 356 803</td><td></td></tr><tr><td>Vein Pick</td><td>Harvard Apparatus, Cambridge, UK</td><td>72-4169</td><td>For single use only</td></tr><tr><td>Saline</td><td>Laboratorium Dr. G. Bichsel AG, , Switzerland</td><td>1330055</td><td></td></tr><tr><td>Polysorb 5-0 suture</td><td>Covidien AG, Switzerland</td><td>UL 202</td><td>Monofilament</td></tr><tr><td>Sulfadoxine and Trimethoprim-Trimethazol</td><td>Werner Stricker AG, Switzerland</td><td>Swissmedic Nr. 50&#039;361</td><td></td></tr><tr><td>Antiseptic- Octenisept</td><td>Sch&amp;uuml;lke &amp;amp; Mayr AG, Switzerland</td><td>GTIN: 4032651214068</td><td></td></tr><tr><td>Phosphate Buffered Saline</td><td>Roth</td><td>1058.1</td><td></td></tr><tr><td>Isobutanol-2-Methylbutane</td><td>Sigma-Aldrich, Switzerland</td><td>M32631-1L</td><td></td></tr><tr><td>Optimum Cutting Temperature compound-Tissue-Tek</td><td>VWR Chemicals, Belgium</td><td>25608-930</td><td></td></tr><tr><td>Cryostat</td><td>Leica, Glattbrugg, Switzerland</td><td>Leica CM1860 UV</td><td></td></tr><tr><td>Glass slide- Superfrost Plus</td><td>Thermo Scientific</td><td>4951PLUS4</td><td></td></tr><tr><td>Mayer&#039;s Haematoxylin</td><td>Sigma-Aldrich, Switzerland</td><td>MHS32-1L</td><td></td></tr><tr><td>Eosin 0.5% aq.</td><td>Sigma-Aldrich, Switzerland</td><td>HT110232-1L</td><td></td></tr><tr><td>Oil Red O</td><td>Sigma-Aldrich, Switzerland</td><td>O0625-25G</td><td></td></tr><tr><td>&amp;alpha;-smooth muscle actin antibody</td><td>Abcam, UK.</td><td>ab7817</td><td></td></tr><tr><td>Macrophage Clone RAM11 antibody</td><td>DAKO, Switzerland</td><td>M063301</td><td></td></tr><tr><td>Hoechst</td><td>Abcam, UK.</td><td>ab145596</td><td></td></tr><tr><td>Goat polyclonal Secondary Antibody (Chromeo 546)</td><td>Abcam, UK.</td><td>ab60316</td><td></td></tr><tr><td>Alexa Fluor 488/547</td><td>Abcam, UK.</td><td></td><td></td></tr><tr><td>Glycergel Mounting Medium, Aqueous</td><td>DAKO, Switzerland</td><td>C056330</td><td></td></tr><tr><td>Hematoxylin for Movat pentachrome staining</td><td>Sigma-Aldrich, Switzerland</td><td>H3136-25G</td><td></td></tr><tr><td>Ferric chloride for Movat pentachrome staining</td><td>Sigma-Aldrich, Switzerland</td><td>157740-100G</td><td></td></tr><tr><td>Iodine for Movat staining</td><td>Sigma-Aldrich, Switzerland</td><td>207772-100G</td><td></td></tr><tr><td>Potassium iodide for Movat pentachrome staining</td><td>Sigma-Aldrich, Switzerland</td><td>60400-100G-F</td><td></td></tr><tr><td>Alcian blue for Movat staining</td><td>Sigma-Aldrich, Switzerland</td><td>A5268-10G</td><td></td></tr><tr><td>Strong Ammonia for Movat pentachrome staining</td><td>Sigma-Aldrich, Switzerland</td><td>320145-500ML</td><td></td></tr><tr><td>Brilliant crocein MOO for Movat pentachrome staining</td><td>Sigma-Aldrich, Switzerland</td><td>210757-50G</td><td></td></tr><tr><td>Acid Fuchsin for Movat pentachrome staining</td><td>Sigma-Aldrich, Switzerland</td><td>F8129-50G</td><td></td></tr><tr><td>Sodium Thiosulfate for Movat pentachrome staining</td><td>Sigma-Aldrich, Switzerland</td><td>72049-250G</td><td></td></tr><tr><td>Phosphotungstic acid for Movat pentachrome staining</td><td>Sigma-Aldrich, Switzerland</td><td>79690-100G</td><td></td></tr><tr><td>Crocin for Movat pentachrome staining</td><td>Sigma-Aldrich, Switzerland</td><td>17304-5G</td><td></td></tr><tr><td>EUKITT for Movat pentachrome staining</td><td>Sigma-Aldrich, Switzerland</td><td>03989-100ML</td><td></td></tr></tbody></table>

the rabbit model of accelerated atherosclerosis: a methodological perspective of the iliac artery balloon injury

급성 관상 동맥 증후군 관상동맥 폐색 동맥 경 화성 플 라크 개발 및 파열에서에서 유래한 공업화 한 세계에서 사망의 주요 원인입니다. 뉴질랜드 화이트 (NZW) 토끼는 동맥 경화 증의 연구를 위한 동물 모델로 서 널리 사용 됩니다. 그들은 개발 atherogenic 다이어트; 먹 일 때 자연 스러운 병 변 그러나,이 4-8 개월의 긴 시간이 필요합니다. 더욱 강화 하 고 가속 atherogenesis, atherogenic 다이어트와 기계적 내 피 상해는 종종 사용 됩니다. 유도 하는 토끼의 동맥 경 화성 플 라크에 대 한 제시 절차 atherogenic 다이어트 먹이 NZW 토끼의 왼쪽된 장 골 동맥에 피를 방해 하는 풍선 카 테 터를 사용 합니다. 풍선 카 테 터로 인 한 이러한 기계적 손상 시간 종속 방식에서 neointimal 지질 축적을 시작 하는 염증 성 반응의 사슬을 유도 합니다. 동맥 경 화성 플 라크 풍선 부상 쇼 neointimal 두껍게 광범위 한 지질 침투와 높은 평활 근 셀 내용, 대 식 세포의 존재에 따라 거품 세포 파생. 이 기술은 간단 하 고, 재현 고 제어 길이 장 골 동맥 내 플 라크를 생성 한다. 모든 절차는 20-30 분 이내 완료 됩니다. 절차는 낮은 사망률과 안전 하며 또한 상당한 내 병 변 취득에 높은 성공. 풍선 카 테 터의 절차 2 주 이내에 동맥 경화 동맥 상해 결과 유도 한다. 질병 병리학을 조사를 위해이 모델을 사용할 수 있습니다 진단 이미징 새로운 치료 전략을 평가 하 고.

장 골 동맥의 풍선 부상 합병증 (그림 1) 없이 성공적으로 수행 되었다. 총 수술 시간 30 분 부상에 대 한 20에서 배열 했다 수행 하나의 장 골 동맥, 및 두 동맥에 대 한 35 ~ 45 분. 토끼 풍선 부상 후 1 시간 이내 복구. 모든 동물 상당한 체중 감소 없이 건강 한 나타났다. 아니 감염, 부 종 또는 동맥 혈전 증 발생 했습니다. 상처 영역 봉합 사이트에서 몇 ?...

Watch this Scientific Journal Video about The Rabbit Model of Accelerated Atherosclerosis: A Methodological Perspective of the Iliac Artery Balloon Injury at JoVE.com

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

동맥 경화 증의 동물 모델 메커니즘을 이해 하 고 패 개발 또는 파열, 산업화 된 세계에서 죽음의 주요 원인이 방지 하기 위해 새로운 접근을 조사 하기 위해 필수적입니다. 이 프로토콜 풍선 부상과 콜레스테롤 풍부한 다이어트의 조합을 사용 하 여 토끼 장 골 동맥에 동맥 경 화성 플 라크를 유도.

가속된 동맥 경화의 토끼 모델: 장 골 동맥 풍선 상해의 방법 론 적 관점

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

the-rabbit-model-accelerated-atherosclerosis-methodological

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

12.0K 조회수.  University of Fribourg. 동맥 경화 증의 동물 모델 메커니즘을 이해 하 고 패 개발 또는 파열, 산업화 된 세계에서 죽음의 주요 원인이 방지 하기 위해 새로운 접근을 조사 하기 위해 필수적입니다. 이 프로토콜 풍선 부상과 콜레스테롤 풍부한 다이어트의 조합을 사용 하 여 토끼 장 골 동맥에 동맥 경 화성 플 라크를 유도.

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

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.

부분 결합하여 마우스 경동맥 동맥에있는 방해 흐름 유발 죽상 경화증과 경동맥 내피로부터 RNA 분리의 간단한 방법의 모델

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

연구

의학

In vivo Near Infrared Fluorescence (NIRF) Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

The vascular response to injury is a well-orchestrated inflammatory response triggered by the accumulation of macrophages within the vessel wall leading to an accumulation of lipid-laden intra-luminal plaque, smooth muscle cell proliferation and progressive narrowing of the vessel lumen. The formation of such vulnerable plaques prone to rupture underlies the majority of cases of acute myocardial infarction. The complex molecular and cellular inflammatory cascade is orchestrated by the recruitment of T lymphocytes and macrophages and their paracrine effects on endothelial and smooth muscle cells.1
Molecular imaging in atherosclerosis has evolved into an important clinical and research tool that allows in vivo visualization of inflammation and other biological processes. Several recent examples demonstrate the ability to detect high-risk plaques in patients, and assess the effects of pharmacotherapeutics in atherosclerosis.4 While a number of molecular imaging approaches (in particular MRI and PET) can image biological aspects of large vessels such as the carotid arteries, scant options exist for imaging of coronary arteries.2 The advent of high-resolution optical imaging strategies, in particular near-infrared fluorescence (NIRF), coupled with activatable fluorescent probes, have enhanced sensitivity and led to the development of new intravascular strategies to improve biological imaging of human coronary atherosclerosis.
Near infrared fluorescence (NIRF) molecular imaging utilizes excitation light with a defined band width (650-900 nm) as a source of photons that, when delivered to an optical contrast agent or fluorescent probe, emits fluorescence in the NIR window that can be detected using an appropriate emission filter and a high sensitivity charge-coupled camera. As opposed to visible light, NIR light penetrates deeply into tissue, is markedly less attenuated by endogenous photon absorbers such as hemoglobin, lipid and water, and enables high target-to-background ratios due to reduced autofluorescence in the NIR window. Imaging within the NIR 'window' can substantially improve the potential for in vivo imaging.2,5
Inflammatory cysteine proteases have been well studied using activatable NIRF probes10, and play important roles in atherogenesis. Via degradation of the extracellular matrix, cysteine proteases contribute importantly to the progression and complications of atherosclerosis8. In particular, the cysteine protease, cathepsin B, is highly expressed and colocalizes with macrophages in experimental murine, rabbit, and human atheromata.3,6,7 In addition, cathepsin B activity in plaques can be sensed in vivo utilizing a previously described 1-D intravascular near-infrared fluorescence technology6, in conjunction with an injectable nanosensor agent that consists of a poly-lysine polymer backbone derivatized with multiple NIR fluorochromes (VM110/Prosense750, ex/em 750/780nm, VisEn Medical, Woburn, MA) that results in strong intramolecular quenching at baseline.10 Following targeted enzymatic cleavage by cysteine proteases such as cathepsin B (known to colocalize with plaque macrophages), the fluorochromes separate, resulting in substantial amplification of the NIRF signal. Intravascular detection of NIR fluorescence signal by the utilized novel 2D intravascular NIRF catheter now enables high-resolution, geometrically accurate in vivo detection of cathepsin B activity in inflamed plaque.
In vivo molecular imaging of atherosclerosis using catheter-based 2D NIRF imaging, as opposed to a prior 1-D spectroscopic approach,6 is a novel and promising tool that utilizes augmented protease activity in macrophage-rich plaque to detect vascular inflammation.11,12 The following research protocol describes the use of an intravascular 2-dimensional NIRF catheter to image and characterize plaque structure utilizing key aspects of plaque biology. It is a translatable platform that when integrated with existing clinical imaging technologies including angiography and intravascular ultrasound (IVUS), offers a unique and novel integrated multimodal molecular imaging technique that distinguishes inflammatory atheromata, and allows detection of intravascular NIRF signals in human-sized coronary arteries.

In vivo Near Infrared Fluorescence NIRF Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

We detail a new near-infrared fluorescence (NIRF) catheter for 2-dimensional intravascular molecular imaging of plaque biology in vivo. The NIRF catheter can visualize key biological processes such as inflammation by reporting on the presence of plaque-avid activatable and targeted NIR fluorochromes. The catheter utilizes clinical engineering and power requirements and is targeted for application in human coronary arteries. The following research study describes a multimodal imaging strategy that utilizes a novel in vivo intravascular NIRF catheter to image and quantify inflammatory plaque in proteolytically active inflamed rabbit atheromata.

염증성 플라크의 적외선 형광 (NIRF) Intravascular 분자 이미징, Multimodal 접근 가까운 생체내에서 죽상 경화증의 영상을

The vascular response to injury is a well-orchestrated inflammatory response triggered by the accumulation of macrophages within the vessel wall leading ...

Inducing Myointimal Hyperplasia Versus Atherosclerosis in Mice: An Introduction of Two Valid Models

Various in vivo laboratory rodent models for the induction of artery stenosis have been established to mimic diseases that include arterial plaque formation and stenosis, as observed for example in ischemic heart disease. Two highly reproducible mouse models&#160;&#8211;&#160;both resulting in artery stenosis but each underlying a different pathway of development&#160;&#8211; are introduced here. The models represent the two most common causes of artery stenosis; namely one mouse model for each myointimal hyperplasia, and atherosclerosis are shown. To induce myointimal hyperplasia, a balloon catheter injury of the abdominal aorta is performed. For the development of atherosclerotic plaque, the ApoE -/- mouse model in combination with western fatty diet is used. Different model-adapted options for the measurement and evaluation of the results are named and described in this manuscript. The introduction and comparison of these two models provides information for scientists to choose the appropriate artery stenosis model in accordance to the scientific question asked.

This video shows two models of intimal plaque development in murine arteries and emphasizes the differences in myointimal hyperplasia and atherosclerosis.

쥐에서 동맥 경화 수스 Myointimal 증식을 유도 : 두 개의 유효한 모델의 소개

Various in vivo laboratory rodent models for the induction of artery stenosis have been established to mimic diseases that include arterial plaque ...

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.

쥐 경동맥의 혈관 풍선 부상과 관내 관리

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 ...

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.

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.

생쥐 경동맥 풍선 손상 모델은 반대로 혈관 리모델링의 치료를 테스트하는

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 ...

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.

마우스에서 가속 죽상 경화증의 유도: "와이어 부상" 모델

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.

쥐 모델에서 Infrarenal 복부 대동맥 클램핑에 의해 재현 동맥 발가 벗김 부상

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

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

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

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

풍선-기반 부상 마우스 복 부 대동맥에서 Myointimal 증식을 유도 하

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

A 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.

쥐 경동맥 압력 제어 세그먼트 풍선 부상 과식 치료 응용 프로그램

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

A Rabbit Aortic Valve Stenosis Model Induced by Direct Balloon Injury

Animal models are emerging as an important tool to understand the pathologic mechanisms underlying aortic valve stenosis (AVS) because of the lack of access to reliable sources of diseased human aortic valves. Among the various animal models, AVS rabbit models are one of the most commonly used in large animal studies. However, traditional AVS rabbit models require a long-term period of dietary supplementation and genetic manipulation to induce significant stenosis in the aortic valve, limiting their use in experimental studies. To address these limitations, a new AVS rabbit model is proposed, in which stenosis is induced by a direct balloon injury to the aortic valve. The present protocol describes a successful technique for inducing AVS in New Zealand white (NZW) rabbits, with step-by-step procedures for the preparation, the surgical procedure, and the post-operative care. This simple and reproducible model offers a promising approach for studying the initiation and progression of AVS and provides a valuable tool for investigating the underlying pathological mechanisms of the disease.

An appropriate animal model is needed to understand the pathologic mechanisms underlying aortic valve stenosis (AVS) and to evaluate the efficacy of therapeutic interventions. The present protocol describes a new procedure for developing the AVS rabbit model via a direct balloon injury in vivo.

직접적인 풍선 손상으로 인한 토끼 대동맥 판막 협착증 모델

Animal models are emerging as an important tool to understand the pathologic mechanisms underlying aortic valve stenosis (AVS) because of the lack of ...