Dr de CAESTECKER laboratuvar NIH 1RO1 HL093057-01 ve 1RC4DK090770-01 tarafından desteklenmektedir. Dr Harris&#39;in laboratuvar DK38226, DK51265, DK62794 ve Gaziler İdaresi fon tarafından desteklenmektedir. Ayrıca Vanderbilt O&#39;Brien Böbrek Sakatlık Merkezi hibe 1P30 DK079341 tarafından sağlanan fare böbrek hasarı cerrahi, serum kreatinin ve fibrozis testleri için destek.

<ol><li> Ameliyat öncesi tüm cerrahi aletler otoklav. Bir farklı fareler üzerinde birden fazla cerrahisine ise, sıcak bir boncuk sterilizatör kullanılarak sterilize sonra kullanımdan sonra durulama ve unutmayın. Bu% 70 etanol emmek için yeterli değildir. </li><li> Ameliyat öncesi 0.5 ml S / C steril serum fizyolojik verin ve hemen ameliyat sonrası ameliyat sırasında vücudun sıvı kaybını telafi etmek için. </li><li> Farenin tartılır. </li><li> IP Xylazine / Ketamin karışımı kullanılarak fare anestezisi. Genellikle cerrahi uçak anestezi ulaşmak için fareler için 3-5 dakika sürer. </li><li> Kesi karışmasını saç tutmak için yeterli sınır bölgesi ile cerrahi alan tıraş. Bu ameliyat olduğu, bir hazırlık alanda değil yapılması gerekmektedir. </li><li> İşlem sırasında kurumasını önlemek için gözlere oftalmik yağlama merhem uygulayın. </li><li> Emici tezgah ped ile kaplı ısıtılmış yüzey üzerinde eğilimli fare yerleştirin. Cerrahi yüzeye Bant ayaklar. </li><li> Kullanıyorsanızsu banyosu sistemi, cerrahi öncesi 38 ° C 1 saat olarak belirlenmiştir ısı. </li><li> Aseptik hazırlık Betadine çözüm çubuk çubuk kullanarak: Betadine kaldırmak için Nolvasan ile ovma ardından her zaman, 3x çevreye doğru sitenin merkezine fırçalayın. Tüm Betadine Nolvasan ile durulayarak kesi önce cerrahi alanından çıkarılmalıdır. Aseptik işlem eldiven başlamadan önce steril olarak değiştirilmelidir. </li><li> Steril cerrahi örtü ile cerrahi alan kapsamaktadır. Bu cerrahi alanında girmesini sokak saç önlemek ve ameliyat sırasında steril aletleri koymak için bir alan sağlayacaktır. </li><li> Deri yoluyla böbrek yeri palpe ediniz. </li><li> Makas ve forseps kullanarak fare (yaklaşık 1.5 cm) orta hat boyunca dorsal cilt kesin. </li><li> Makas ve forseps kullanarak künt diseksiyon ile bu kesiden sol ve sağ dorsal tarafın üzerinden ayrı cilt ve cilt altı katmanları. </li><li> Sağ fl ile küçük bir kesi yapmakkas ve böbrek üzerinde fasya ank ve sağ böbrek exteriorize. </li><li> Dikkatlice çevre dokulara ücretsiz böbrek üst ve alt kutuplar incelemek. Üst kutup yerinden yağ dokusu böbrek kapalı itilebilir ama belli ki fare (Şekil 1) çıkarılmamalıdır kendi kan akımı taşıyan böbreküstü bezi içerdiğini unutmayın. Çevre dokudan özgürleştirici böbrek sonra, bir çift cerrahi düğüm kullanarak sağ böbrek hilusu etrafında 4-0 ipek sütür kravat. Siteye kesilecek görselleştirmek böylece düğüm distalinde kesim olarak böbrek tutmak için uzun dikiş ~ 2 bir ucunu bırakın. Steril serum fizyolojik ile güdük yıkayınız. </li><li> Emilebilir sütür kullanarak kas tabakası kapatın. </li><li> Sol kanattan kas ve böbrek üzerinde fasya üzerinden küçük bir kesi yapmak ve sol böbrek (Şekil 2A) exteriorize. </li><li> Dikkatle böbrek bırakırken künt forseps kullanarak böbrek tutunforseps (Şekil 2B-2F) kullanarak yağ dokusu çevreleyen pedikül. Çalışmaları için &quot;sahte&quot; kontrolleri tasarlarken bir ihtiyacı bu hatırlamak çok künt forseps kullanarak böbrek üzerinde çok fazla baskı renal hasar neden olabilir. Eğer sahte kontrol renal hasar (serum kreatinin bir artış kanıtladığı) gelişmekte fark ederseniz, alternatif bir yöntem renal pedikül çevreleyen yağ dokusu serbest bırakmak için düşünebilirsiniz hafifçe tuzlu ıslatılmış steril ile dokulara rulo için pamuk- uçlu çubuklar. Deneyimlerimiz yağ oldukça sıkı renal pedikül bağlı ve kolayca pamuklu ile kaldırılamaz, bu nedenle bu kullanmak isteyeyim ilk yaklaşım olmayabilir. Ancak, bu modelin diğer yönleri gibi, bu uygulama ve deneyim ile kendiniz için kurmak zorunda olacak bir şeydir. </li><li> Tutma forseps (kelepçe yerleştirme kolaylaştırmak için) kullanarak olmayan travmatik damar kelepçe sol renal pedikül Kelepçe, farklı CLA için zamanlayıcı kullanınmp kez fare zorlanma ve (bkz. Tablo 1) gerekli yaralanma şiddetine bağlı olarak. </li><li> Cilt (Şekil 2H) ile sol böbrek Kapak. </li><li> Tuzlu batırılmış gazlı bez ile cilt kesisi Kapak. </li><li> Böbrek exteriorize, düzgün esmer görünüm onaylamak ve belirtilen zaman sonra kelepçe serbest bırakın ve bu esmer görünüm (Şekil 2I / G) böbrek boyunca düzgün tersine onaylayın. Böbrek hızlı ve düzgün yeniden serpmek (pembe kadar) fare daha fazla analiz dışında tutulmalıdır yoksa. Yavaşça retroperitoneal uzaya geri böbrek itin. </li><li> Emilebilir sütür kullanarak kas tabakası kapatın. </li><li> Monofilament naylon emilmeyen sütür kullanarak cilt katmanı kapatın. </li><li> Buprenorfin analjezi yönetmek. </li><li> Onlar uyanmak kadar bir ısıtma yastığı üzerine uzakta cerrahi alanında fareler aktarın. </li><li> Hayvan odasına fareler dönün. </li><li> Yakından fareler izlemek ve ek doz vermekBuprenorfinin olarak ağrı veya rahatsızlık her 8-12 saat yukarıda belirtilen. Analjezik yönetiminde çıkan klinik belirtiler depresyon veya diğer davranış değişiklikleri, anormal görünüm veya duruş gibi pilo-ereksiyon olarak, kambur duruş, bakım ve beslenme eksikliği, ya da hareketsizlik içerir. </li></ol>

<ol>
	<li>Singh, A. P., Junemann, A., Muthuraman, A., Jaggi, A. S., Singh, N., Grover, K., Dhawan, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+of+acute+renal+failure.">Animal models of acute renal failure.</a> Pharmacological reports: PR. 64, 31-44 (2012).</li><li>Heyman, S. N., Rosenberger, C., Rosen, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+ischemia-reperfusion:+Biases+and+myths-the+proximal+vs.+Distal+hypoxic+tubular+injury+debate+revisited.">Experimental ischemia-reperfusion: Biases and myths-the proximal vs. Distal hypoxic tubular injury debate revisited.</a> Kidney Int. 77, 9-16 (2010).</li><li>Lieberthal, W., Nigam, S. 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+renal+failure.+II.+Experimental+models+of+acute+renal+failure:+Imperfect+but+indispensable.">Acute renal failure. II. Experimental models of acute renal failure: Imperfect but indispensable.</a> Am. J. Physiol. Renal Physiol. 278, F1-F12 (2000).</li><li>Dunn, S. R., Qi, Z., Bottinger, E. P., Breyer, M. D., Sharma, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Utility+of+endogenous+creatinine+clearance+as+a+measure+of+renal+function+in+mice.">Utility of endogenous creatinine clearance as a measure of renal function in mice.</a> Kidney Int. 65, 1959-1967 (2004).</li><li>Yuen, P. S., Dunn, S. R., Miyaji, T., Yasuda, H., Sharma, K., Star, R. A. <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+method+for+HPLC+determination+of+creatinine+in+mouse+serum.">A simplified method for HPLC determination of creatinine in mouse serum.</a> Am. J. Physiol. Renal Physiol. 286, F1116-F1119 (2004).</li><li>Hetu, P. O., Gingras, M. E., Vinet, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+validation+of+a+rapid+liquid+chromatography+isotope+dilution+tandem+mass+spectrometry+(LC-IDMS/MS)+method+for+serum+creatinine.">Development and validation of a rapid liquid chromatography isotope dilution tandem mass spectrometry (LC-IDMS/MS) method for serum creatinine.</a> Clin. Biochem. 43, 1158-1162 (2010).</li><li>Keppler, A., Gretz, N., Schmidt, R., Kloetzer, H. M., Groene, H. J., Lelongt, B., Meyer, M., Sadick, M., Pill, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Plasma+creatinine+determination+in+mice+and+rats:+An+enzymatic+method+compares+favorably+with+a+high-performance+liquid+chromatography+assay.">Plasma creatinine determination in mice and rats: An enzymatic method compares favorably with a high-performance liquid chromatography assay.</a> Kidney Int. 71, 74-78 (2007).</li><li>Yang, L., Besschetnova, T. Y., Brooks, C. R., Shah, J. V., Bonventre, J. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epithelial+cell+cycle+arrest+in+G2/M+mediates+kidney+fibrosis+after+injury.">Epithelial cell cycle arrest in G2/M mediates kidney fibrosis after injury.</a> Nat. Med. 16, 535-543 (2010).</li><li>Shanley, P. F., Rosen, M. D., Brezis, M., Silva, P., Epstein, F. H., Rosen, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Topography+of+focal+proximal+tubular+necrosis+after+ischemia+with+reflow+in+the+rat+kidney.">Topography of focal proximal tubular necrosis after ischemia with reflow in the rat kidney.</a> Am. J. Pathol. 122, 462-468 (1986).</li><li>Chawla, L. S., Amdur, R. L., Amodeo, S., Kimmel, P. L., Palant, C. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+severity+of+acute+kidney+injury+predicts+progression+to+chronic+kidney+disease.">The severity of acute kidney injury predicts progression to chronic kidney disease.</a> Kidney Int. 79, 1361-1369 (2012).</li><li>Lo, L. J., Go, A. S., Chertow, G. M., McCulloch, C. E., Fan, D., Ordonez, J. D., Hsu, C. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dialysis-requiring+acute+renal+failure+increases+the+risk+of+progressive+chronic+kidney+disease.">Dialysis-requiring acute renal failure increases the risk of progressive chronic kidney disease.</a> Kidney Int. 76, 893-899 (2009).</li><li>Burne, M. J., Haq, M., Matsuse, H., Mohapatra, S., Rabb, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+susceptibility+to+renal+ischemia+reperfusion+injury+revealed+in+a+murine+model.">Genetic susceptibility to renal ischemia reperfusion injury revealed in a murine model.</a> Transplantation. 69, 1023-1025 (2000).</li><li>Muller, V., Losonczy, G., Heemann, U., Vannay, A., Fekete, A., Reusz, G., Tulassay, T., Szabo, A. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sexual+dimorphism+in+renal+ischemia-reperfusion+injury+in+rats:+Possible+role+of+endothelin.">Sexual dimorphism in renal ischemia-reperfusion injury in rats: Possible role of endothelin.</a> Kidney Int. 62, 1364-1371 (2002).</li><li>Schmitt, R., Marlier, A., Cantley, L. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Zag+expression+during+aging+suppresses+proliferation+after+kidney+injury.">Zag expression during aging suppresses proliferation after kidney injury.</a> J. Am. Soc. Nephrol. 19, 2375-2383 (2008).</li><li>Oxburgh, L., de Caestecker, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ischemia-reperfusion+injury+of+the+mouse+kidney.">Ischemia-reperfusion injury of the mouse kidney.</a> Methods Mol. Biol. 886, 363-379 (2012).</li></ol>

Yazarlar ifşa hiçbir çıkar çatışması var.

Biz orta ve şiddetli renal hasar etkilerini incelemek için IR-AKI iki model açıklar. Bu modeller bize düşük mortalite ile tutarlı ve öngörülebilir hasara yol sağlar. Bizim protokol geleneksel olarak bu model ile ilgili zorluklar ve tuzaklar birçok özetliyor. Ayrıca, renal pedikül sıkma uzunluğuna bağlı olarak, model eksik kurtarma ve kalıcı böbrek fibrozis ile büyük ölçüde geri dönüşümlü hafif ve orta derecede AKI, ya da daha şiddetli AKI neden olduğunu göstermiştir. Ciddi IR-AKI mod...

Akut böbrek hasarı (ABH) deneysel modellerinde çeşitli insan durumu (yeni, kapsamlı bir yorum 1 için referans bakınız) çeşitliliği ve karmaşıklığı maç için geliştirilmiştir. Bu modellerin her biri kendi güçlü ve zayıf yönleri vardır, ve her çeşitli verimliliği ile ilgili insan koşulları taklit ederken, hiçbiri tam olarak insan meslektaşları patofizyolojisi modeli. Iskemi reperfüzyon (IR)-kaynaklı AKI kemirgenlerde akut iskemi kaynaklı böbrek hasarının bir model olarak geliştirilmiştir. Bu modelde görülen renal tübüler hasar şiddeti nadiren sınırlamalara rağmen, renal hipoperfüzyon yaralanmaları olan hastalarda 2 görülmektedir ve büyük ölçüde çünkü bu model nispeten tekrarlanabilir doğa, geniş kullanım sağlamıştır ve bu beklenirken, devam edecek ortak temel AKI mekanizmaları, onarım, ve tedavi 3 birçok içine önemli bilgiler sağlamak için. IR cerrahi fam gerektirir, Şekil 1 &#39;de, basitleştirilmiş bir formda gösterilen adres fare böbrek anatomisi ile iliarity. Böbrek iskemi-reperfüzyon (IR) hasarı cerrahi ventral (laparatomi) veya dorsal (retroperitoneal) yaklaşımları ile yapılabilir. Daha az travmatik olduğundan daha hızlı iyileşme süreleri ve sağkalım (ilk işlem öğrenme özellikle) sağlayan, bir dorsal yaklaşım kullanın. Böbrek IR yaralanma tek taraflı veya iki taraflı yapılabilir. Bununla birlikte, etkili bir kelepçe basınçları (kelepçe çene arasına perihiler yağ araya girmesi sonucu olabilir) ve / veya yanlar arasında renal iskemi yanıtları arasında önemli farklılıklar daha değişken sonuçlara yol açar. Bu aşılamaz bir sorun olmasa da, bu model için önemli bir konu olan, deneyler arasında değişkenlik artırabilir. Tek taraflı IR kontra-yan nefrektomi ile yapılabilir. Bu da pediküller arasında sıkma değişkenlik azalttığı ve aynı zamanda seçim bizim yöntemdirllows tek başına tek taraflı IR ile etkilenmez böbrek fonksiyon, değerlendirmek. Farelerde böbrek fonksiyonu değerlendirmek için en pratik vs uygun yöntem ne olduğu hakkında tartışmalar vardır. Kan üre azotu (BUN) böbrek fonksiyon bir ölçü sağlar ve IR yaralanma da dahil olmak üzere AKI bazı modellerde, yararlı bir &quot;ilk bakış&quot; belirtecidir. Gecikmiş kurtarma gün sonrası cerrahi bir dizi için sıvı içirin azaltır Ancak, BUN seviyeleri, özellikle ventral yaklaşım IR yaralanma sonrası, etkilenebilir farelerin birim durumu etkilenebilir. Serum kreatinin az hidrasyon durumu etkilenir ama açıkça kas kütlesi etkilenir. Serum kreatinin ölçümleri ile ilgili zorluklardan biri pikrik asit bazlı teknikler kullanılarak fare serumu olmayan kromojenler kreatinin tespiti ile ilgili sorunlar olmuştur. Alternatif olarak, merkezleri bir dizi bu eser 4 etkilenmez fare kreatinin ölçmek için bir HPLC-tabanlı bir yöntem geliştirdik. Bununla birlikte, UNLike BUN ve sadece serum 5-10 ul gerektirir pikrik asit, kreatinin deneyleri, HPLC tahlilleri ~ 25 ul analiz başına, serum, iki kopya halinde gerçekleştirilir halinde analiz başına tam kan ~ 100 ul gerektirir. gerektirir Bu fare çalışmalar için sınırlayıcı olabilir. Bazı merkezlerde küçük örnek hacmi 5, 6 analizi izin daha duyarlı HPLC ve kütle spektrometresi tabanlı yöntemler geliştirdik. Ancak, bu teknolojilerin yaygın olarak mevcut değildir. Bir alternatif, enzimatik kaskad tahlili (serum sadece 5-10 ul gerektirir), fare ve sıçan serum numuneleri ile değerlendirilir ve yakından serum kreatinin pikrik asit testleri her zaman aşırı tahmin kreatinin değeri ise 7 HPLC ölçümleri paralel olduğu gösterilmiştir. Bu tahlilde yaygın AKI literatürde kullanılan olmasa da, tahlil, kullanımı basit, ticari olarak mevcuttur, ve farelerde IR indüklenen AKI Bu modelle güvenilir sonuçlar veren.

<table border="1">
	<tbody>
		<tr>
			<td>Drug or solution</td>
			<td>Company</td>
			<td>Catalogue number</td>
			<td>Comments</td>
		</tr>
		<tr>
			<td>Ketamine (anesthetic)</td>
			<td>Webster Veterinary</td>
			<td>07-881-9413 (100 mg/ml)</td>
			<td>Final concentration 30 mg/ml Ketamine with 3 mg/ml Xylazine administered at 3-4 ml/kg (100 mg/kg Ketamine and 10 mg/kg Xylazine)</td>
		</tr>
		<tr>
			<td>Xylazine (analgesic)</td>
			<td>Webster Veterinary</td>
			<td>07-808-1939 (100 mg/ml)</td>
			<td>As above</td>
		</tr>
		<tr>
			<td>Antisedan (anti-Xylazine)</td>
			<td>Webster Veterinary</td>
			<td>07-867-7097 (5 mg/ml)</td>
			<td>Optional: to reverse effects of Xylazine in order to increase the rate of recovery after surgery Use 0.5-0.7 mg/kg</td>
		</tr>
		<tr>
			<td>Buprenorphine (analgesic)</td>
			<td>Bedford Laboratories</td>
			<td>NDC 55390-100-10 (0.3 mg/ml)</td>
			<td>Dose 0.05-0.1 &mu;g/g every 8-12 hr. Make up a 1:5 dilution in sterile normal saline preoperatively and give 2.5-5 &mu;l/g of the diluted solution.</td>
		</tr>
		<tr>
			<td>Ophthalmic ointment</td>
			<td>Dechra</td>
			<td>NDC 17033-211-38</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Betadine swab sticks</td>
			<td>Purdue Products L.P.</td>
			<td>NDC 67618-153-01</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Nolvasan Surgical Scrub</td>
			<td>Pfizer Animal Care</td>
			<td>300253</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Table 2. Drugs and solutions for I/R surgery.</td>
		</tr>
		<tr>
			<td>Iris Scissors</td>
			<td>Integra Miltex</td>
			<td>VWR 21909-404</td>
			<td>For skin and muscle dissection</td>
		</tr>
		<tr>
			<td>Curved iris forceps</td>
			<td>FST</td>
			<td>11065-07</td>
			<td>To hold skin and muscle, to remove peri-nephric fat</td>
		</tr>
		<tr>
			<td>Blunt forceps</td>
			<td>ROBOZ</td>
			<td>RS-5228</td>
			<td>To manipulate kidneys</td>
		</tr>
		<tr>
			<td>Vascular clamp (795 g pressure)</td>
			<td>ROBOZ</td>
			<td>RS-5459</td>
			<td>Good for C57BL/6 and BALB/c mice strains</td>
		</tr>
		<tr>
			<td>Schwartz clip applying forceps</td>
			<td>ROBOZ</td>
			<td>RS-5450</td>
			<td>For RS-5459 clamps</td>
		</tr>
		<tr>
			<td>Vascular clamp (75-85 g pressure)</td>
			<td>FST</td>
			<td>18055-02</td>
			<td>Optimal for CD1 mice strain, but also can be used for C57BL/6 and BALB/c</td>
		</tr>
		<tr>
			<td>Micro-Serrefine Clamp Applicator-14 cm with Lock</td>
			<td>FST</td>
			<td>18056-14</td>
			<td>For FST 18055-02 clamps</td>
		</tr>
		<tr>
			<td>Gayman T/Pump (water-bath heated platform)</td>
			<td>Braintree Scientific</td>
			<td>TP-650</td>
			<td>The water heater is set at 38 &deg;C 1 hr prior surgery</td>
		</tr>
		<tr>
			<td>15" W x 24" Heated platform</td>
			<td>Braintree Scientific</td>
			<td>HHP-2</td>
			<td>Connect to Gayman heated water pump</td>
		</tr>
		<tr>
			<td>Heating pad</td>
			<td>Braintree Scientific</td>
			<td>Model 39DP</td>
			<td>Uses microwave for heating. This is useful for warming during recovery</td>
		</tr>
		<tr>
			<td>Absorbable suture (Vicryl 5-0)</td>
			<td>ETHICON</td>
			<td>VCP834G</td>
			<td>For fascia and muscle</td>
		</tr>
		<tr>
			<td>Non-absorbable suture: monofilament nylon MONOMID 6/0 660B)</td>
			<td>CP Medical</td>
			<td>CP-B660B-03</td>
			<td>For skin</td>
		</tr>
		<tr>
			<td>Halsey needle holder: Tungsten Carbide/13 cm</td>
			<td>FST</td>
			<td>12501-13</td>
			<td>To hold suture needles</td>
		</tr>
		<tr>
			<td>Timer</td>
			<td>FST</td>
			<td>06-662-3</td>
			<td>To quantify ischemia time</td>
		</tr>
		<tr>
			<td>Hot bead sterilizer</td>
			<td>FST</td>
			<td>18000-45</td>
			<td>To sterilize instruments between mice if you are performing multiple surgeries (it is not sufficient to soak instruments in ethanol)</td>
		</tr>
		<tr>
			<td>Electric razor</td>
			<td>Braintree Scientific</td>
			<td>CLP-22965</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Tape</td>
			<td>Durapore</td>
			<td>1538-0</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>1 ml Syringe</td>
			<td>EXELint</td>
			<td>26044</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Sterile cotton tipped applicators</td>
			<td>Kendall</td>
			<td>8884541300</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Absorbent BenchPad</td>
			<td>VWR</td>
			<td>56617-014</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Surgical drapes</td>
			<td>VWR</td>
			<td>21902-985</td>
			<td>Need to be autoclaved</td>
		</tr>
		<tr>
			<td>2'' x 2'' Gauze Pads</td>
			<td>Medi-First</td>
			<td>60673</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Sterile gloves</td>
			<td>Cardinal Health</td>
			<td>2D7251</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Table 3. Equipment and instruments for I/R surgery.</td>
		</tr>
	</tbody>
</table>

ischemia-reperfusion model of acute kidney injury and post injury fibrosis in mice

Iskemi-reperfüzyon oluşturulan akut böbrek hasarı (IR-AKI) yaygın olarak farelerde AKI bir model olarak kullanılan, ancak sonuçlar genellikle analiz yanıltabilecek yüksek, sık sık rapor edilmemiş ölüm oranları ile oldukça değişkendir edilir. Bilateral renal pedikül sıkma yaygın IR-AKI ikna etmek için kullanılır, ama etkili kelepçe basınçları ve / veya böbrek arasında iskemi böbrek yanıtları arasındaki farklılıkları daha sık değişken sonuçlara yol olduğunu. Buna ek olarak, daha kısa kelepçe kat daha fazla değişken boru hasara yol bilinmektedir, ve daha uzun kelepçe kez ikili yaralanma geçiren fareler daha tutarlı boru hasarına yol ise, genellikle şiddetli böbrek yetmezliği nedeniyle yaralanma sonrası ilk 3 gün içinde ölür. Sonrası yaralanma hayatta kalma geliştirmek ve daha tutarlı ve öngörülebilir sonuçlar elde etmek için, biz taraf nefrektomi ardından tek taraflı iskemi-reperfüzyon hasarının iki model geliştirdik. Her iki ameliyat f sonuçlanan cerrahi stresi azaltarak, bir dorsal yaklaşım kullanılarak yapılmaktadıryaygın olarak fare IR-AKI ameliyatlar için kullanılan rom ventral laparotomi,. Orta derecede yaralanmaya BALB / c farelerde indüksiyonu için tek taraflı 26 dakika boyunca renal pedikül sıkma geçmesi ve aynı zamanda eş zamanlı kontralateral nefrektomi. Bu yaklaşımı kullanarak, farelerin% 50-60 yaralanma sonrası orta AKI 24 saat geliştirmek ama farelerin% 90-100 hayatta. Daha şiddetli AKI ikna etmek için, BALB / c farelerde 8 gün yaralanma sonrası kontralateral nefrektomi ardından 30 dakika için terminal böbrek pedikül tabi. Bu 90-100% hayatta kalma ile yaralanma sonrası böbrek iyileşme fonksiyonel değerlendirme sağlar. Erken sonrası yaralanma tübüler hasar yanı sıra sonrası yaralanma fibrozis bu modeli kullanarak son derece tutarlıdır.

Aynı anda karşı taraf nefrektomi ile Orta IR bağlı AKI. Tek taraflı IR ölçüde sonuçlarında değişkenlik azaltır, ancak bu işlem hayatta kalmak için fareler için gerekli kısa kelepçe kez, biz hala farelerin sadece% 50-60 beklenen böbrek yetmezliği 24 saat geliştirdiği bulundu sonrası travma (Şekil 3A). Çalışmaların zaman böbrek fonksiyonu değerlendirmek ve böbrek yetmezliği gelişmez fareler atmak için izin, yaralanma sonrası en az 24 saat başlatılabil...

Watch this Scientific Journal Video about Ischemia-reperfusion Model of Acute Kidney Injury and Post Injury Fibrosis in Mice at JoVE.com

Ischemia-reperfusion Model of Acute Kidney Injury and Post Injury Fibrosis in Mice

Biz fareler tabi olan orta ve ağır iskemi-reperfüzyon bağlı böbrek hasarı modelleri tanımlamak tek taraflı renal pedikül sırasıyla, eş zamanlı veya gecikmeli taraf nefrektomi takip sıkma. Bu modeller sürekli böbrek fonksiyon bozukluğu ve post-yaralanma fibrozis yol, ancak yaralanma şiddeti ve yaşam fare arka plan, yaş ve cerrahi ekipman bağlıdır.

Farelerde Akut Böbrek Hasarı ve Post Yaralanma Fibrozis iskemi-reperfüzyon Model

Ischemia-reperfusion induced acute kidney injury (IR-AKI) is widely used as a model of AKI in mice, but results are often quite variable with high, often ...

ischemia-reperfusion-model-acute-kidney-injury-post-injury-fibrosis

Research

JoVE Journal

Medicine

39.2K Görüntüleme.  Vanderbilt University Medical Center. Biz fareler tabi olan orta ve ağır iskemi-reperfüzyon bağlı böbrek hasarı modelleri tanımlamak tek taraflı renal pedikül sırasıyla, eş zamanlı veya gecikmeli taraf nefrektomi takip sıkma. Bu modeller sürekli böbrek fonksiyon bozukluğu ve post-yaralanma fibrozis yol, ancak yaralanma şiddeti ve yaşam fare arka plan, yaş ve cerrahi ekipman bağlıdır.

Video: Ischemia-reperfusion Model of Acute Kidney Injury and Post Injury Fibrosis in Mice

Normothermic Cardiac Arrest and Cardiopulmonary Resuscitation: A Mouse Model of Ischemia-Reperfusion Injury

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
frequently caused by whole-body hypoperfusion.5,6 Successful reproduction of whole-body hypoperfusion in rodent models has been fraught with 
difficulty.7-9,9,10 Models which employ focal ischemia have repeatedly demonstrated results which do not translate to the clinical setting, and larger 
animal models which allow for whole body hypoperfusion lack access to the full toolset of genetic manipulation possible in the mouse.11,12 However, in recent 
years a mouse model of cardiac arrest and cardiopulmonary resuscitation has emerged which can be adapted to model AKI.13 This model reliably reproduces 
physiologic, functional, anatomic, and histologic outcomes seen in clinical AKI, is rapidly repeatable, and offers all of the significant advantages of a murine surgical 
model, including access to genetic manipulative techniques, low cost relative to large animals, and ease of use. Our group has developed extensive experience 
with use of this model to assess a number of organ-specific outcomes in AKI.14,15

A powerful model for perioperative and critical care related acute kidney injury is presented. Using whole body hypoperfusion induced by cardiac arrest it is possible to nearly replicate the histologic and functional changes of clinical AKI.

Normotermik Kardiyak Arrest ve Kardiyopulmoner Resusitasyon: İskemi-Reperfüzyon Hasarı Fare Modeli

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
frequently caused ...

Tıp

Murine Model of Intestinal Ischemia-reperfusion Injury

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, hypotension, necrotizing enterocolitis, bowel transplantation, trauma and chronic inflammation. Intestinal ischemia-reperfusion (IR) injury is a consequence of acute mesenteric ischemia, caused by inadequate blood flow through the mesenteric vessels, resulting in intestinal damage. Reperfusion following ischemia can further exacerbate damage of the intestine. The mechanisms of IR injury are complex and poorly understood. Therefore, experimental small animal models are critical for understanding the pathophysiology of IR injury and the development of novel therapies.
Here we describe a mouse model of acute intestinal IR injury that provides reproducible injury of the small intestine without mortality. This is achieved by inducing ischemia in the region of the distal ileum by temporally occluding the peripheral and terminal collateral branches of the superior mesenteric artery for 60 min using microvascular clips. Reperfusion for 1 hr, or 2 hr after injury results in reproducible injury of the intestine examined by histological analysis. Proper position of the microvascular clips is critical for the procedure. Therefore the video clip provides a detailed visual step-by-step description of this technique. This model of intestinal IR injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

Here we describe the detailed procedure of intestinal ischemia-reperfusion in mice which results in reproducible injury without mortality to encourage the standardization of this technique across the field. This model of intestinal ischemia-reperfusion injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

Bağırsak İskemi-reperfüzyon Hasarı Murin Modeli

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, ...

A Mouse Model of Retinal Ischemia-Reperfusion Injury Through Elevation of Intraocular Pressure

Retinal ischemia-reperfusion (I/R) is a pathophysiological process contributing to cellular damage in multiple ocular conditions, including glaucoma, diabetic retinopathy, and retinal vascular occlusions. Rodent models of I/R injury are providing significant insights into mechanisms and treatment strategies for human I/R injury, especially with regard to neurodegenerative damage in the retinal neurovascular unit. Presented here is a protocol for inducing retinal I/R injury in mice through elevation of intraocular pressure (IOP). In this protocol, the ocular anterior chamber is cannulated with a needle, through which flows the drip of an elevated saline reservoir. Using this drip to raise IOP above systolic arterial blood pressure, a practitioner temporarily halts inner retinal blood flow (ischemia). When circulation is reinstated (reperfusion) by removal of the cannula, severe cellular damage ensues, resulting ultimately in retinal neurodegeneration. Recent studies demonstrate inflammation, vascular permeability, and capillary degeneration as additional elements of this model. Compared to alternative retinal I/R methodologies, such as retinal arterial ligation, retinal I/R injury by elevated IOP offers advantages in its&#160;anatomical specificity, experimental tractability, and technical accessibility, presenting itself as a valuable tool for examining neuronal pathogenesis and therapy in the retinal neurovascular unit.

This article describes a procedure for inducing retinal ischemia-reperfusion injury by elevated intraocular pressure in mice. Retinal ischemia-reperfusion injury by elevated intraocular pressure serves to model human pathologies characterized by compromised oxygen and nutrient delivery in the retina, enabling researchers to examine potential cellular mechanisms and treatments for human diseases of the retinal neurovascular unit.

Göz İçi Basıncı Yükselmesi ile Retina İskemi-Reperfüzyon Hasarı Fare Modeli

Retinal ischemia-reperfusion (I/R) is a pathophysiological process contributing to cellular damage in multiple ocular conditions, including glaucoma, ...

Evaluation of Coronary Flow Reserve After Myocardial Ischemia Reperfusion in Rats

Coronary artery disease is the leading cause of death worldwide. After an acute myocardial infarction, early and successful myocardial intervention via recanalization of the coronary artery is the most effective strategy for reducing the size of ischemic myocardium. The coronary microvasculature cannot be visualized and imaged&#160;in vivo, but there are several invasive and noninvasive techniques that can be used to assess parameters which depend directly on coronary microvascular function. The endothelial function after ischemia reperfusion can be assessed also at the level of the coronary circulation via the coronary flow reserve (CFR).&#160;In this study, peak velocity of left anterior descending (LAD) coronary arteries was measured in rats in vivo via Transthoracic Doppler Echocardiography during resting and stress challenge (induced by Dobutamine). A normal heart can increase its coronary blood flow up to four times above the resting values during stress induction. Following ischemia reperfusion, we found a significantly diminished CFR, which can be used as a marker of coronary microvascular dysfunction. CFR has opened a window on the importance of microvascular dysfunction and has been shown to predict cardiovascular risk independent of whether the severe obstructive disease is present.

Coronary flow reserve (CFR), is defined as the ratio of maximal coronary blood flow to the resting coronary blood flow. We present a protocol for evaluating CFR in rats via ultrasound, which offers the opportunity to predict cardiovascular risk factors in the absence of obstructive coronary disease.

Farelerde miyokardiyal Iskemi reperfüzyon sonrası koroner akış rezervi değerlendirilmesi

Coronary artery disease is the leading cause of death worldwide. After an acute myocardial infarction, early and successful myocardial intervention via ...

A Pre-clinical Rat Model for the Study of Ischemia-reperfusion Injury in Reconstructive Microsurgery

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many areas of biomedical research due to its cost-effectiveness and its translation to humans. This protocol describes a method to create a preclinical free skin flap model in rats with ischemia-reperfusion injury. The described 3 cm x 6 cm rat free skin flap model is easily obtained after the placement of several vascular ligatures and the section of the vascular pedicle. Then, 8 h after the ischemic insult and completion of the microsurgical anastomosis, the free skin flap develops the tissue damage. These ischemia-reperfusion injury-related damages can be studied in this model, making it a suitable model for evaluating therapeutic agents to address this pathophysiological process. Furthermore, two main monitoring techniques are described in the protocol for the assessment of this animal model: transit-time ultrasound technology and laser speckle contrast analysis.

Here, we describe a pre-clinical animal model for studying the pathophysiology of ischemia-reperfusion injury in reconstructive microsurgery. This free skin flap model based on the superficial caudal epigastric vessels in the rat may also allow for the evaluation of different therapies and compounds to counteract ischemia-reperfusion injury-related damage.

Rekonstrüktif Mikrocerrahide İskemi-Reperfüzyon Hasarı Nın İnceleştirilmesinde Klinik Öncesi Sıçan Modeli

Ischemia-reperfusion injury is the main cause of flap failure in reconstructive microsurgery. The rat is the preferred preclinical animal model in many ...

Delayed Intramyocardial Delivery of Stem Cells after Ischemia Reperfusion Injury in a Murine Model

There is significant interest in the use of stem cells (SCs) for the recovery of cardiac function in individuals with myocardial injuries. Most commonly, cardiac stem cell therapy is studied by delivering SCs concurrently with the induction of myocardial injury. However, this approach presents two significant limitations: the early hostile pro-inflammatory ischemic environment may affect the survival of transplanted SCs, and it does not represent the subacute infarction scenario where SCs will likely be used. Here we describe a two-part series of surgical procedures for the induction of ischemia-reperfusion injury and delivery of mesenchymal stem cells (MSCs). This method of stem cell administration may allow for the longer viability and retention around damaged tissue by circumventing the initial immune response. A model of ischemia reperfusion injury was induced in mice accompanied by the delivery of mesenchymal stem cells (3.0 x 105), stably expressing the reporter gene firefly luciferase under the constitutively expressed CMV promoter, intramyocardially 7 days later. The animals were imaged via ultrasound and bioluminescent imaging for confirmation of injury and injection of cells, respectively. Importantly, there was no added complication rate when performing this two-procedure approach for SC delivery. This method of stem cell administration, collectively with the utilization of state-of-the-art reporter genes, may allow for the in vivo study of viability and retention of transplanted SCs in a situation of chronic ischemia commonly seen clinically, while also circumventing the initial pro-inflammatory response. In summary, we established a protocol for the delayed delivery of stem cells into the myocardium, which can be used as a potential new approach in promoting regeneration of the damaged tissue.

Stems cells are continuously investigated as potential treatments for individuals with myocardial damage, however, their decreased viability and retention within injured tissue can impact their long-term efficacy. In this manuscript we describe an alternative method for stem cell delivery in a murine model of ischemia reperfusion injury.

Bir Murine Modelinde İskemi Reperfüzyon Yaralanması Sonrası Kök Hücrelerin Gecikmiş İntramiyokardiyal Teslimi

There is significant interest in the use of stem cells (SCs) for the recovery of cardiac function in individuals with myocardial injuries. Most commonly, ...

Acute Kidney Injury Model Induced by Cisplatin in Adult Zebrafish

Cisplatin is commonly used as chemotherapy. Although it has positive effects in cancer-treated individuals, cisplatin can easily accumulate in the kidney due to its low molecular weight. Such accumulation causes the death of tubular cells and can induce the development of Acute Kidney Injury (AKI), which is characterized by a quick decrease in kidney function, tissue damage, and immune cells infiltration. If administered in specific doses cisplatin can be a useful tool as an AKI inducer in animal models. The zebrafish has appeared as an interesting model to study renal function, kidney regeneration, and injury, as renal structures conserve functional similarities with mammals. Adult zebrafish injected with cisplatin shows decreased survival, kidney cell death, and increased inflammation markers after 24 h post-injection (hpi). In this model, immune cells infiltration and cell death can be assessed by flow cytometry and TUNEL assay. This protocol describes the procedures to induce AKI in adult zebrafish by intraperitoneal cisplatin injection and subsequently demonstrates how to collect the renal tissue for flow cytometry processing and cell death TUNEL assay. These techniques will be useful to understand the effects of cisplatin as a nephrotoxic agent and will contribute to the expansion of AKI models in adult zebrafish. This model can also be used to study kidney regeneration, in the search for compounds that treat or prevent kidney damage and to study inflammation in AKI. Moreover, the methods used in this protocol will improve the characterization of tissue damage and inflammation, which are therapeutic targets in kidney-associated comorbidities.

This protocol describes the procedures to induce Acute Kidney Injury (AKI) in adult zebrafish using cisplatin as a nephrotoxic agent. We detailed the steps to evaluate the reproducibility of the technique and two techniques to analyze inflammation and cell death in the renal tissue, flow cytometry and TUNEL, respectively.

Yetişkin Zebra Balıklarında Cisplatin tarafından İndüklenen Akut Böbrek Yaralanması Modeli

Cisplatin is commonly used as chemotherapy. Although it has positive effects in cancer-treated individuals, cisplatin can easily accumulate in the kidney ...

Organ Ischemia-Reperfusion Injury by Simulating Hemodynamic Changes in Rat Liver Transplant Model

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, including ischemia-reperfusion injury (IRI) of extrahepatic organs. This model requires numerous experiments and devices. The duration of anhepatic phase is closely related to the time to develop IRI after transplantation. In this experiment, we used hemodynamic changes to induce extrahepatic organ damage in rats and determined the maximum tolerance time. The time until the most severe organ injury varied for different organs. This method can easily be replicated and can also be used to study IRI of the extrahepatic organs after liver transplantation.

This paper provides a detailed description of how to build an animal model of the anhepatic phase (liver ischemia) in rats to facilitate basic research into ischemia-reperfusion injury after liver transplantation.

Sıçan Karaciğer Nakli Modelinde Hemodinamik Değişiklikleri Simüle Ederek Organ İskemi-Reperfüzyon Yaralanması

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, ...

An Effective Mouse Model of Unilateral Renal Ischemia-Reperfusion Injury

Ischemia-reperfusion injury (IRI) is the leading cause of acute renal failure and is a significant contributor to delayed graft function. Animal models are the only available resources that mimic the complexities of the IRI-associated damage encountered in vivo. This paper describes an effective mouse model of unilateral renal IRI that delivers highly reproducible data. Ischemia is induced by occluding the right renal pedicle for 30 min followed by reperfusion. In addition to the surgical procedure, a sequential overview of the expected physiological and histopathological changes following renal IRI will be provided by comparing data from seven different reperfusion times (4 h, 8 h, 16 h, 1 day, 2 days, 4 days, and 7 days). Critical data for planning experiments ahead, such as mean surgical time, average anesthetic consumption, and body weight changes over time, will be shared. This work will help researchers implement a reliable renal IRI model and select the appropriate reperfusion time that aligns with their intended investigative goals.

Renal ischemia-reperfusion injury is associated with high morbidity and mortality in hospitalized patients. Here, we present a simple and effective mouse model of unilateral renal ischemia-reperfusion injury and provide a sequential overview of representative pathological changes observed in the kidney.

Tek Taraflı İskemi-Reperfüzyon Hasarının Etkili Bir Fare Modeli

Ischemia-reperfusion injury (IRI) is the leading cause of acute renal failure and is a significant contributor to delayed graft function. Animal models ...

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

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

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

Farelerde Miyokard Enfarktüsü ve Miyokard İskemi-Reperfüzyon Hasarının İndüksiyonu

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