Yazarlar, teknik yardım için Christiane Pahrmann teşekkür ederim. SS Deutsche Forschungsgemeinschaft (DFG) (SCHR992/3-2 ve SCHR992/4-2) fon aldı.

Tüm hayvan çalışmaları ilgili hayvan bakım kurallarına göre yapılmalıdır. Önce başlangıç ​​protokole hayvan çalışmaları için kurumsal onay almak. 1.. Miyointimal Büyümesi Modeli (A) Intimal hiperplazi modeli için, C57Bl/6J (Stok sayısı 000.664, C57BI/6J) yaklaşık 25 gr ağırlığındaki 8 haftalık yaşta fareler satın. , Alışılmış koşullar altında, bu deneyler için Ev farenin; fareler standart fare yemi yem ve su ad libitum sağlar. <ol><li> Fare hazırlanması: Bir indüksiyon odası kullanan izofluran (% 2) ile anestezi durumuna fare tanıtılması. <ol><li> Sırtında fare yattı ve anestezi sağlamak için yüz maskesi ile burun ve ağız kapağı, saç düzeltici kullanılarak karın saç çıkarın. </li><li> Provo-İyot ile, iki döngüleri,% 80 etanol ile takip evrensel karın bölgesi, dezenfekte. </li><li> Yokluğunun sağlayınanestezi yeterli derinliği izlemek için arka uzuv kısma reflekslerin. </li><li> Karın organları ortaya çıkarmak için iki adımda linea alba boyunca deri ve kas ayırın. </li><li> Bir tuzlu nemli eldiven bağırsakları yatıyordu. Nemli tutmak için bağırsakların etrafında eldiven sarın. </li><li> Dikkatle abdominal aorta taneleştirici yağ dokusu kaldırmak. Herhangi bir şube damarları kesmemeye özen, aşağı onun çatallanma infrarenal aort Açığa. </li><li> İlk olarak, kan akışını kapatmaya, proksimal üzerinde infrarenal aorta bir mikrocerrahi kelepçe yerleştirin. Şimdi, aort bifürkasyon yanında ikinci bir uzak kelepçe geçerlidir. </li><li> Kan akışı kesildiğinde, proksimal kelepçe yakın aort içine küçük bir kesi yapar. </li><li> % 0.9 serum fizyolojik ile nemlendirilmelidir olmuştur balon uçlu bir kateter, yerleştirin ve aort distal kelepçe doğru ilerlemek. </li><li> Endotel hasarı için, Balloo şişirilerek aort soymakn dikkatlice ve yavaşça yakın kelepçenin yönde kateter çekerek. </li><li> Iki denüdasyon manevrayı tekrarlayın. </li><li> Kateteri çıkarın ve 10-0 prolen çalışan suturlerle aort kesi kapatın. </li><li> Dikkatlice distal kelepçe açın. Aortotomy yerinde kanama durumunda, gerekli, tekrar kelepçeyi kapatmak kanamayı bulmak ve ek kesintiye dikiş yerleştirin. </li><li> Kanama görülmedi edilebilir, yavaş yavaş proksimal kelepçe açın. </li><li> Aort nabız kesiden distale görünür olmalıdır. Şimdi bağırsak yerinde geri düzenlenebilir. </li><li> Önceden ısıtılmış steril tuzlu su kullanan karın boşluğuna durulayın. </li><li> Karın duvarı 6-0 prolen çalışan suturlerle kas tabakası ile başlayan kapatın. </li><li> Sonraki 5-0 prolen performans çalışan sütür kullanım altında cildi adapte. </li><li> Süre boyunca fare hala uyuşturulduktan, bir deri altı inje yoluyla 4-5 mg / kg Carprofen uygulayınction. </li><li> Ağrı kontrolü için içme suyu (100 ml başına 50 mg metamizol) için metamizolün ekleyin ve 3 gün sonrası ameliyat için devam ediyor. Kurtarma boyunca hayvan izleyin. Not: Bu model için gözlem süresi 28 gündür. </li></ol></li></ol> 2.. Ateroskleroz Modeli (B) 4 haftalık bir yaşta fareler (Stock sayısı 002052, B6.129P2-Apoetm1Unc / J), ve daha sonra 4-6 ay boyunca beslenen ateroskleroz modeli için, apolipoprotein E-eksikli (- / - ApoE) satın. , Alışılmış koşullar altında, bu deneyler için Ev farenin; farelere girişte yüksek lipid batı diyet (Harlan Laboratories TD.88137) beslemek ve 4 ila 6 aylık bir süre için su ad libitum sağlar. <ol><li> Fare hazırlama: <ol><li> ApoE Yem - / - Batı diyet ile fareler 4 hafta başlayan. Diyet manipülasyon girişte doğrudan başlamalıdır. </li><li> Deney süresi boyunca batı diyet sürdürmek. &lt;/ Li&gt; </ol></li></ol> 3. Analizi <ol><li> Miyointimal hiperplazi modeli (A) Bu modelde ilgi bölgesi &quot;lezyon alanı&quot; dir. Analizi için seçenekler luminal obliterasyonu arasında, I / E oranı (intima / media oranı), maksimal myointima kalınlık, miyointimal bölge, fibrozit trikrom ve mononükleer hücreler için H &amp; E boyama (Şekil 1A-1C &#39;de gösterildiği gibi). Bilimsel soruya göre Picrosirius kollajen için kırmızı, immünhistokimya ve konfokal yöntemleri gibi çok sayıda diğer boyamaları uygulanabilir. </li><li> Ateroskleroz modeli (B) Bu modelde ilgi bölgeleri, aort kapak, aort hem de azalan aorta bulunmaktadır. Analiz için Seçenekler: Sudan kırmızı lipidler için boyama ve aort kapağı, aort kavsi ve inen aorta (Şekil 1D gösterildiği gibi) bir trichrome boyama. Bu model aynı zamanda boyama gibi diğer çeşitli seçenekler elde edilirNötr lipidler, Picrosirius kırmızı, immünohistokimya ve konfokal yöntemleri için örneğin Yağlı Kırmızı O için. <ol><li> Sudan kırmızı boyama: taze hasat aort dokusu üzerine Sudan kırmızı boyanma gerçekleştirin. <ol><li> Hasat öncesi diyet 6s çıkarın. </li><li> Bir indüksiyon odası kullanan izofluran (% 2) ile anestezi durumuna fare tanıtılması. Anestezi derinliğinin izlenmesi için yeterli bir arka uzuv kısma refleksleri yokluğunu sağlayın. </li><li> Göğüs açın; Kalbin tepe kesilmiş ve aort düzeltmek için aort kökü ile% 4 PFA (3 mi), ardından tuzlu su ilk (3 mi) ile sol ventrikil yoluyla serpmek. </li><li> Onun çatallanma kökünden aort ücretsiz. Yağ dokusu mümkün olduğunca çıkarmaya çalışın. </li><li> Kalp ve aort kökü kesilmiş ve histoloji için% 4 PFA saklayın. </li><li> Histoloji için aort kökü dikkatlice AOR kesilmesini sağlamak için bir dik konumda parafin blok içine yerleştirilebilir zorundadır3 kapak broşürler gösterilecek tik kök üreten slaytlar. </li><li> Aort ağacın bir ucunu kesin ve ince iğneler ile silikon tabaka üzerine bunu düzeltmek. Uzunlamasına açın ve aort aort doku çözmek. </li><li> % 50 etanol ile doku durulayın. Yaklaşık 15 dakika boyunca Sudan III boyama çözeltisi ile doku bırakın. </li><li> Fazla Sudan III çıkarmak için% 50 etanol ile hafifçe durulayın. DH 2 O ile durulayın, ve daha fazla analiz için fotoğraf çekmek: (% 50 etanol uzak boyayı olabilir. Not). </li></ol></li></ol></li></ol>

<ol>
	<li>Manuel, D. G., Leung, M., Nguyen, K., Tanuseputro, P., Johansen, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Burden+of+cardiovascular+disease+in+Canada.">Burden of cardiovascular disease in Canada.</a> Can J Cardiol. 19, 997-1004 (2003).</li><li>ER, O. B., Ma, X., Simard, T., Pourdjabbar, A., Hibbert, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathogenesis+of+neointima+formation+following+vascular+injury.">Pathogenesis of neointima formation following vascular injury.</a> Cardiovasc Hematol Disord Drug Targets. 11, 30-39 (2011).</li><li>Ross, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+pathogenesis+of+atherosclerosis:+a+perspective+for+the+1990s.">The pathogenesis of atherosclerosis: a perspective for the 1990s.</a> Nature. 362, 801-809 (1993).</li><li>Meng, X., Zhang, F., Valji, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasound+guidance+for+the+creation+of+a+small+animal+model+of+aortic+injury.">Ultrasound guidance for the creation of a small animal model of aortic injury.</a> J Vasc Interv Radiol. 22, 1193-1197 (2011).</li><li>Deuse, T., Koyanagi, T., Erben, R. G., 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=Sustained+inhibition+of+epsilon+protein+kinase+C+inhibits+vascular+restenosis+after+balloon+injury+and+stenting.">Sustained inhibition of epsilon protein kinase C inhibits vascular restenosis after balloon injury and stenting.</a> Circulation. 122, 170-178 (2010).</li><li>Schrepfer, S., Deuse, T., Sultan, K. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibition+of+restenosis+development+after+mechanical+injury:+a+new+field+of+application+for+malononitrilamides.">Inhibition of restenosis development after mechanical injury: a new field of application for malononitrilamides.</a> Cardiology. 108, 128-137 (2007).</li><li>Tao, M., Mauro, C. R., Yu, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simplified+murine+intimal+hyperplasia+model+founded+on+a+focal+carotid+stenosis.">A simplified murine intimal hyperplasia model founded on a focal carotid stenosis.</a> Am J Pathol. 182, 277-287 (2013).</li><li>Kumar, A., Lindner, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Remodeling+with+neointima+formation+in+the+mouse+carotid+artery+after+cessation+of+blood+flow.">Remodeling with neointima formation in the mouse carotid artery after cessation of blood flow.</a> Arterioscler Thromb Vasc Biol. 17, 2238-2244 (1997).</li><li>Hua, X., Deuse, T., Michelakis, E. D., 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=Human+internal+mammary+artery+(IMA)+transplantation+and+stenting:+a+human+model+to+study+the+development+of+in-stent+restenosis.">Human internal mammary artery (IMA) transplantation and stenting: a human model to study the development of in-stent restenosis.</a> J Vis Exp. , 3663 (2012).</li><li>Daugherty, A., Whitman, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantification+of+atherosclerosis+in+mice.">Quantification of atherosclerosis in mice.</a> Methods Mol Biol. , (2003).</li><li>Nakashima, Y., Plump, A. S., Raines, E. W., Breslow, J. L., Ross, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ApoE-deficient+mice+develop+lesions+of+all+phases+of+atherosclerosis+throughout+the+arterial+tree.">ApoE-deficient mice develop lesions of all phases of atherosclerosis throughout the arterial tree.</a> Arterioscler Thromb. 14, 133-1340 (1994).</li><li>Zhang, Y., Li, L., You, J., Cao, J., Fu, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+7-difluoromethyl-5,+4'-dimethoxygenistein+on+aorta+atherosclerosis+in+hyperlipidemia+ApoE(-/-)+mice+induced+by+a+cholesterol-rich+diet.">Effect of 7-difluoromethyl-5, 4'-dimethoxygenistein on aorta atherosclerosis in hyperlipidemia ApoE(-/-) mice induced by a cholesterol-rich diet.</a> Drug Des Devel Ther. 7, 233-242 (2013).</li><li>Rudolph, T. K., Rudolph, V., Edreira, M. 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=Nitro-fatty+acids+reduce+atherosclerosis+in+apolipoprotein+E-deficient+mice.">Nitro-fatty acids reduce atherosclerosis in apolipoprotein E-deficient mice.</a> Arterioscler Thromb Vasc Biol. 30, 938-9345 (2010).</li><li>Rondelet, B., Kerbaul, F., Motte, S., 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=Bosentan+for+the+prevention+of+overcirculation-induced+experimental+pulmonary+arterial+hypertension.">Bosentan for the prevention of overcirculation-induced experimental pulmonary arterial hypertension.</a> Circulation. 107, 1329-1335 (2003).</li><li>Zeibig, S., Li, Z., Wagner, S., 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=Effect+of+the+oxLDL+binding+protein+Fc-CD68+on+plaque+extension+and+vulnerability+in+atherosclerosis.">Effect of the oxLDL binding protein Fc-CD68 on plaque extension and vulnerability in atherosclerosis.</a> Circ Res. 108, 695-703 (2011).</li><li>Painter, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Myointimal+hyperplasia:+pathogenesis+and+implications.+2.+Animal+injury+models+and+mechanical+factors.">Myointimal hyperplasia: pathogenesis and implications. 2. Animal injury models and mechanical factors.</a> Artificial organs. 15, 103-118 (1991).</li><li>Nitschke, Y., Weissen-Plenz, G., Terkeltaub, R., Rutsch, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Npp1+promotes+atherosclerosis+in+ApoE+knockout+mice.">Npp1 promotes atherosclerosis in ApoE knockout mice.</a> Journal of cellular and molecular. 15, 2273-2283 (2011).</li></ol>

Yazarlar ifşa hiçbir şey yok.

Her iki hastalık, hastanın içindeki belirtileri neden olmakla birlikte, plak gelişimi altında yatan mekanizmaları ve bu nedenle tedavi yaklaşımları 2 çok farklıdır. Arteriyel darlık farklı şekillerde hastalarda klinik bulgular yanı sıra plak gelişim zaman dilimi yatan mekanizma bağlıdır. Hastada klinik bulgulara bağlı olarak, numune farklı analiz yöntemleri 5,15 yapılmalıdır. Bu, seçilen model 14 tarafından taklit edilmektedir ha...

Sanayileşmiş ülkelerde iskemik kalp hastalığı mortalite 1 önde gelen nedenidir. Koroner arter darlığı için nedenleri çeşitlidir ve en yaygın tedavi stratejisini kalan 2 revaskülarizasyon ile miyointimal hiperplazi yanı sıra ateroskleroz içerir. Açık intimal hiperplazi ve ateroskleroz mekanistik yollar arasında ayrım Research modelleri arter plak gelişimi patabiyolojik ve patofizyolojik süreçleri araştırmak için gereklidir. Bu video miyointimal hiperplazi veya ateroskleroz gelişimi ya da çalışma için kullanılan iki farklı fare modelleri yerleştirilir. Mıyoıntimal hiperplazının ilk ateroskleroz 3 için anlatılan &quot;cevabı-to-zedelenmesi&quot; paradigması ile oluşturulması için kabul edilmektedir. Endotel tabakasının mekanik bozulma parakrin etkisi ile gelişmiş yoğun bir yeniden yol açar. Birkaç di vardırfferent hayvan modelleri yaygın mıyoıntimal hiperplazının incelemek için kullanılmıştır. Bazı gruplar sıçanların 4 aortta metal cihazlar kullanın. Bir balon kateter aort soyulma modeli aynı zamanda yaygın laboratuar sıçanlarda 5,6 kullanılır. Laboratuvar fareleri 7 gerçekleştirilen intimal hiperplazi modeli karotis arter ligasyonu uygular ve miyointimal lezyonlar 8 uyarmaktadır. Laboratuvarda, abdominal aort denüdasyon modeli - yanı sıra bir insanlaşmış stent restenoz modeli 9 sık kullanılan - miyointimal hiperplazi gelişimini incelemek için. Bu video uygun deneysel hayvan modeli seçerek arter darlığının mekanik veya patofizyolojik çalışmalar için çok önemli olduğunu vurguluyor. Miyointimal hiperplazi modelleri ateroskleroz modellerden ayırt edilmelidir. İkincisi, apolipoprotein E-eksikli (ApoE - / -) western diyet ile kombinasyon halinde farenin yaygın Atheroscl üretmek için kullanılırerotik lezyonlar 10,11,12,13. Farelerde miyointimal hastalığı bu tür her ikisinin de indüksiyonu için örnek olarak damar darlığı 14 analiz etmek için adapte edilmiş farklı model seçenekleri ile birlikte, burada gösterilmektedir.

<table border="0" cellpadding="0" cellspacing="0" style="width:617px;" width="618">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="17">
			<td height="17" style="height:17px;width:156px;">Name</td>
			<td style="width:183px;">Company</td>
			<td style="width:139px;">Catalog number</td>
			<td style="width:140px;">Comments</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Thyoglycollate Broth 3%</td>
			<td>Fluka</td>
			<td align="right">70157</td>
			<td>powder</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">PFA 4%</td>
			<td>Electron Microscopy Sciences</td>
			<td>#157135S</td>
			<td align="right">20%</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Sudan III staining solution</td>
			<td>Sigma Aldrich</td>
			<td>S4131</td>
			<td>powder</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">mouse C57BL/6J</td>
			<td>Jackson Laboratories</td>
			<td>&#160;Stock # 0006664</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">mouse ApoE-/-</td>
			<td>Jackson Laboratories</td>
			<td>&#160;Stock #002052</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Western Diet</td>
			<td>Harlan Laboratories</td>
			<td>TD.88137</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">hair clipper</td>
			<td>WAHL</td>
			<td>8786-451A ARCO SE</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Forene</td>
			<td>Abbott</td>
			<td></td>
			<td>Isoflurane</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">microsurgical clamp</td>
			<td>Fine Science Tools</td>
			<td>18055-04</td>
			<td>Micro-Serrefine - 4mm</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">clamp applicator</td>
			<td>Fine Science Tools</td>
			<td>18056-14</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">catheter</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">10-0 prolene suture</td>
			<td>Ethicon</td>
			<td>788G</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">6-0 prolene suture</td>
			<td>Ethicon</td>
			<td>8709H</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">5-0 prolene suture</td>
			<td>Ethicon</td>
			<td>EH7229H</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Rimadyl</td>
			<td>Pfizer&#160;</td>
			<td></td>
			<td>Carprofen</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Metacam 1.5mg/ml</td>
			<td>Boehringer Ingelheim</td>
			<td></td>
			<td>Metamizol</td>
		</tr>
	</tbody>
</table>

inducing myointimal hyperplasia versus atherosclerosis in mice: an introduction of two valid models

Arter stenozu uyarılması için çeşitli in vivo laboratuvar kemirgen modelleri, iskemik kalp hastalığı, örneğin görüldüğü gibi, arter plak oluşumu ve stenoz hastalıkları taklit etmek için kurulmuştur. İki yüksek tekrarlanabilir fare modelleri - iki arter darlığı sonuçlanan ancak her gelişme farklı bir yol yatan - burada tanıtılmaktadır. Modeller arter darlığının en yaygın iki nedeni temsil eder; yani tek bir fare, her miyointimal hiperplazisi için model ve ateroskleroz gösterilmiştir. Miyointimal hiperplaziyi indükleme için, abdominal aortun balon kateter yaralanma gerçekleştirilir. Aterosklerotik plak gelişimi için, ApoE - / - Batı yağlı diyet ile birlikte fare modeli kullanılmaktadır. Sonuçlarının ölçülmesi ve değerlendirilmesi için farklı model seçenekleri adapte bu yazının adlandırılmış ve tanımlanmıştır. Bu iki modelin tanıtımı ve karşılaştırma informat sağlarBilim adamları sordu bilimsel soruya göre uygun arter darlığı modelini seçmek için iyon.

Şekil 1, farklı analiz seçenekleri ve her iki model için uyarılan hastalık durumunu gösterir. Intimal hiperplazi modeli için, trikrom boyanması için temsili kesitlerin analizi gösterilmektedir. Bu kesitler, ben / M oranı, en plak kalınlığı, lümen maksimum intimal kalınlıkta yüzde olarak luminal yok edilmesi, hem de leke alanı gibi sonuçlar ölçülebilir ve hesaplanabilir. Ateroskleroz modeli sonucunu değerlendirmek için, yukarıda adı geçen tüm y?...

Watch this Scientific Journal Video about Inducing Myointimal Hyperplasia Versus Atherosclerosis in Mice: An Introduction of Two Valid Models at JoVE.com

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

Bu video kemirgen arterlerde intimal plak iki kalkınma modelleri gösterir ve miyointimal hiperplazi ve ateroskleroz farklılıkları vurgulamaktadır.

Farelerde Ateroskleroz Karşı miyointimal hiperplazide uyaran: İki Geçerli Modellerinin An Introduction

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

inducing-myointimal-hyperplasia-versus-atherosclerosis-mice-an

Research

JoVE Journal

Medicine

12.0K Görüntüleme.  University Hospital Hamburg.  Bu video kemirgen arterlerde intimal plak iki kalkınma modelleri gösterir ve miyointimal hiperplazi ve ateroskleroz farklılıkları vurgulamaktadır. 

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

Implantation of a Carotid Cuff for Triggering Shear-stress Induced Atherosclerosis in Mice

It is widely accepted that alterations in vascular shear stress trigger the expression of inflammatory genes in endothelial cells and thereby induce atherosclerosis (reviewed in 1 and 2). The role of shear stress has been extensively studied in vitro investigating the influence of flow dynamics on cultured endothelial cells 1,3,4 and in vivo in larger animals and humans 1,5,6,7,8. However, highly reproducible small animal models allowing systematic investigation of the influence of shear stress on plaque development are rare. Recently, Nam et al. 9 introduced a mouse model in which the ligation of branches of the carotid artery creates a region of low and oscillatory flow. Although this model causes endothelial dysfunction and rapid formation of atherosclerotic lesions in hyperlipidemic mice, it cannot be excluded that the observed inflammatory response is, at least in part, a consequence of endothelial and/or vessel damage due to ligation.
In order to avoid such limitations, a shear stress modifying cuff has been developed based upon calculated fluid dynamics, whose cone shaped inner lumen was selected to create defined regions of low, high and oscillatory shear stress within the common carotid artery 10. By applying this model in Apolipoprotein E (ApoE) knockout mice fed a high cholesterol western type diet, vascular lesions develop upstream and downstream from the cuff. Their phenotype is correlated with the regional flow dynamics 11 as confirmed by in vivo Magnetic Resonance Imaging (MRI) 12: Low and laminar shear stress upstream of the cuff causes the formation of extensive plaques of a more vulnerable phenotype, whereas oscillatory shear stress downstream of the cuff induces stable atherosclerotic lesions 11. In those regions of high shear stress and high laminar flow within the cuff, typically no atherosclerotic plaques are observed.
 
In conclusion, the shear stress-modifying cuff procedure is a reliable surgical approach to produce phenotypically different atherosclerotic lesions in ApoE-deficient mice.

The constricting cuff presented in this article is designed to induce atherosclerosis in the murine common carotid artery. Due to the conical shape of its inner lumen the implanted cuff generates well-defined regions of low, high and oscillatory shear stress triggering the development of atherosclerotic lesions of different inflammatory phenotypes.

Fare Shear stres İndüklenmiş Ateroskleroz Tetikleme Karotis Kol İmplantasyonu

It is widely accepted that alterations in vascular shear stress trigger the expression of inflammatory genes in endothelial cells and thereby induce ...

Tıp

Quantitative Analysis and Characterization of Atherosclerotic Lesions in the Murine Aortic Sinus

Atherosclerosis is a disease of the large arteries and a major underlying cause of myocardial infarction and stroke. Several different mouse models have been developed to facilitate the study of the molecular and cellular pathophysiology of this disease. In this manuscript we describe specific techniques for the quantification and characterization of atherosclerotic lesions in the murine aortic sinus and ascending aorta. The advantage of this procedure is that it provides an accurate measurement of the cross-sectional area and total volume of the lesion, which can be used to compare atherosclerotic progression across different treatment groups. This is possible through the use of the valve leaflets as an anatomical landmark, together with careful adjustment of the sectioning angle. We also describe basic staining methods that can be used to begin to characterize atherosclerotic progression. These can be further modified to investigate antigens of specific interest to the researcher. The described techniques are generally applicable to a wide variety of existing and newly created dietary and genetically-induced models of atherogenesis.

We describe procedures to quantify and characterize atherosclerotic lesions in mouse models using precision sectioning of the aortic sinus and ascending aorta combined with histochemical and immunohistochemical analysis.

Atherosclerosis is a disease of the large arteries and a major underlying cause of myocardial infarction and stroke. Several different mouse models have ...

Acute Brain Trauma in Mice Followed By Longitudinal Two-photon Imaging

Although acute brain trauma often results from head damage in different accidents and affects a substantial fraction of the population, there is no effective treatment for it yet. Limitations of currently used animal models impede understanding of the pathology mechanism. Multiphoton microscopy allows studying cells and tissues within intact animal brains longitudinally under physiological and pathological conditions. Here, we describe two models of acute brain injury studied by means of two-photon imaging of brain cell behavior under posttraumatic conditions. A selected brain region is injured with a sharp needle to produce a trauma of a controlled width and depth in the brain parenchyma. Our method uses stereotaxic prick with a syringe needle, which can be combined with simultaneous drug application. We propose that this method can be used as an advanced tool to study cellular mechanisms of pathophysiological consequences of acute trauma in mammalian brain in vivo. In this video, we combine acute brain injury with two preparations: cranial window and skull thinning. We also discuss advantages and limitations of both preparations for multisession imaging of brain regeneration after trauma.

Acute brain trauma is a severe injury that has no adequate treatment to date. Multiphoton microscopy allows studying longitudinally the process of acute brain trauma development and probing therapeutical strategies in rodents. Two models of acute brain trauma studied with in vivo two-photon imaging of brain are demonstrated in this protocol.

Boyuna İki foton görüntüleme ile takip Farelerde Akut Beyin Travma

Although acute brain trauma often results from head damage in different accidents and affects a substantial fraction of the population, there is no ...

Minimal Invasive Surgical Procedure of Inducing Myocardial Infarction in Mice

Myocardial infarction still remains the main cause of death in western countries, despite considerable progress in the stent development area in the last decades. For clarification of the underlying mechanisms and the development of new therapeutic strategies, the availability of valid animal models are mandatory. Since we need new insights into pathomechanisms of cardiovascular diseases under in vivo conditions to combat myocardial infarction, the validity of the animal model is a crucial aspect. However, protection of animals are highly relevant in this context. Therefore, we establish a minimally invasive and simple model of myocardial infarction in mice, which assures a high reproducibility and survival rate of animals. Thus, this models fulfils the requirements of the 3R principle (Replacement, Refinement and Reduction) for animal experiments and assure the scientific information needed for further developing of therapeutical strategies for cardiovascular diseases.

A highly reproducible model for myocardial infarction in mice with minimal invasive manipulations is described. The model can be easily performed, resulting in a high reproducibility and survival rate. Thus, the described model will reduce the number of required animals as requested by the 3R principle (Replacement, Refinement and Reduction).

Farelerde Miyokard İnfarktüsü Oluşturan Minimal İnvaziv Cerrahi Prosedür

Myocardial infarction still remains the main cause of death in western countries, despite considerable progress in the stent development area in the last ...

Surgical Models of Gastroesophageal Reflux with Mice

Multiple surgical procedures have been reported to induce gastroesophageal reflux in animals. Herein, we report three surgical models with mice aiming to induce reflux of gastric contents, duodenal contents or mixed contents. Surgical procedures and general principles have been described in detail. A researcher with surgical experience should be able to grasp the technique after a short period of practice. After surgery, most mice can survive and develop reflux esophagitis similar to that in humans. However, it should be noted that histological differences between mouse and human esophagus are the inherent limitations of these surgical models. If used for research on Barrett&#8217;s esophagus and adenocarcinoma, these procedures may need to be combined with genetic modifications.

This article demonstrates surgical procedures of gastroesophageal reflux with mice. These models are useful tools for research on mechanisms and treatment of gastroesophageal reflux disease and potentially Barrett&#8217;s esophagus and esophageal adenocarcinoma.

Fare ile Gastroözofageal Reflü Cerrahi Modelleri

Multiple surgical procedures have been reported to induce gastroesophageal reflux in animals. Herein, we report three surgical models with mice aiming to ...

An Improved Method for Rapid Intubation of the Trachea in Mice

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity is a commonly used model to study both acute lung injury and fibrosis, and multiple methods have been developed for administering bleomycin (and other toxic agents) into the lungs. However, many of these approaches, such as transtracheal instillation, have inherent drawbacks, including the need for strong anesthetics and survival surgery. This paper reports a quick, reproducible method of intratracheal intubation that involves mild inhaled anesthesia, visualization of the trachea, and the use of a surrogate spirometer to confirm exposure. As a proof of concept, 8-12 week old C57BL/6 mice were administered either 2.0 U/kg of bleomycin or an equivalent volume of PBS, and both damage and fibrotic endpoints were measured post-exposure. This procedure allows researchers to treat a large cohort of mice in a relatively short period with little expense and minimal post-procedure care.

This article presents a rapid and simple method for administering bleomycin directly into the mouse trachea via intubation. Key advantages of this method are that it is highly reproducible, easy to master, and does not require specialized equipment or lengthy recovery times.

Farelerde Trakea hızlı intübasyon için geliştirilmiş bir yöntem

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity ...

Glaucoma-inducing Procedure in an In Vivo Rat Model and Whole-mount Retina Preparation

Glaucoma is a disease of the central nervous system affecting retinal ganglion cells (RGCs). RGC axons making up the optic nerve carry visual input to the brain for visual perception. Damage to RGCs and their axons leads to vision loss and/or blindness. Although the specific cause of glaucoma is unknown, the primary risk factor for the disease is an elevated intraocular pressure. Glaucoma-inducing procedures in animal models are a valuable tool to researchers studying the mechanism of RGC death. Such information can lead to the development of effective neuroprotective treatments that could aid in the prevention of vision loss. The protocol in this paper describes a method of inducing glaucoma - like conditions in an in vivo rat model where 50 &#181;l of 2 M hypertonic saline is injected into the episcleral venous plexus. Blanching of the vessels indicates successful injection. This procedure causes loss of RGCs to simulate glaucoma. One month following injection, animals are sacrificed and eyes are removed. Next, the cornea, lens, and vitreous are removed to make an eyecup. The retina is then peeled from the back of the eye and pinned onto sylgard dishes using cactus needles. At this point, neurons in the retina can be stained for analysis. Results from this lab show that approximately 25% of RGCs are lost within one month of the procedure when compared to internal controls. This procedure allows for quantitative analysis of retinal ganglion cell death in an in vivo rat glaucoma model.

Glaucoma-inducing Procedure in an In Vivo Rat Model and Whole-mount Retina Preparation

Glaucoma is characterized by damage to retinal ganglion cells. Inducing glaucoma in animal models can provide insight into the study of this disease. Here, we outline a procedure that induces loss of RGCs in an in vivo rat model and demonstrates the preparation of whole-mount retinas for analysis.

Glokom uyaran bir de Prosedürü<em&gt; İn Vivo</em&gt; Rat Modeli ve Tüm montaj Retina Hazırlık

Glaucoma is a disease of the central nervous system affecting retinal ganglion cells (RGCs). RGC axons making up the optic nerve carry visual input to the ...

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.

Induction of Accelerated Atherosclerosis in Mice: The &quot;Wire-Injury&quot; Model

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.

Farelerde Hızlandırılmış Ateroskleroz Indüksiyonu: "Tel-Yaralanma" Modeli

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

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.

Fare Abdominal aort Myointimal Hiperplazi ikna etmek için balon tabanlı yaralanma

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 ...

Inducing Acute Lung Injury in Mice by Direct Intratracheal Lipopolysaccharide Instillation

Airway administration of lipopolysaccharide (LPS) is a common way to study pulmonary inflammation and acute lung injury (ALI) in small animal models. Various approaches have been described, such as the inhalation of aerosolized LPS as well as nasal or intratracheal instillation. The presented protocol describes a detailed step-by-step procedure to induce ALI in mice by direct intratracheal LPS instillation and perform FACS analysis of blood samples, bronchoalveolar lavage (BAL) fluid, and lung tissue. After intraperitoneal sedation, the trachea is exposed and LPS is administered via a 22 G venous catheter. A robust and reproducible inflammatory reaction with leukocyte invasion, upregulation of proinflammatory cytokines, and disruption of the alveolo-capillary barrier is induced within hours to days, depending on the LPS dosage used. Collection of blood samples, BAL fluid, and lung harvesting, as well as the processing for FACS analysis, are described in detail in the protocol. Although the use of the sterile LPS is not suitable to study pharmacologic interventions in infectious diseases, the described approach offers minimal invasiveness, simple handling, and good reproducibility to answer mechanistic immunological questions. Furthermore, dose titration as well as the use of alternative LPS preparations or mouse strains allow modulation of the clinical effects, which can exhibit different degrees of ALI severity or early vs. late onset of disease symptoms.

Presented here is a step-by-step procedure to induce acute lung injury in mice by direct intratracheal lipopolysaccharide instillation and to perform FACS analysis of blood samples, bronchoalveolar lavage fluid, and lung tissue. Minimal invasiveness, simple handling, good reproducibility, and titration of disease severity are advantages of this approach.

Doğrudan Intratracheal Lipopolysaccharide Instilasyon ile farelerde akut akciğer hasarının İndükleme

Airway administration of lipopolysaccharide (LPS) is a common way to study pulmonary inflammation and acute lung injury (ALI) in small animal models. ...