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>
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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><tbody><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 &amp;mu;g/g every 8-12 hr. Make up a 1:5 dilution in sterile normal saline preoperatively and give 2.5-5 &amp;mu;l/g of the diluted solution.</td></tr><tr><td>Ophthalmic ointment</td><td>Dechra</td><td>NDC 17033-211-38</td><td>&amp;nbsp;</td></tr><tr><td>Betadine swab sticks</td><td>Purdue Products L.P.</td><td>NDC 67618-153-01</td><td>&amp;nbsp;</td></tr><tr><td>Nolvasan Surgical Scrub</td><td>Pfizer Animal Care</td><td>300253</td><td>&amp;nbsp;</td></tr><tr><td colspan="4">&lt;strong&gt;Table 2. Drugs and solutions for I/R surgery.&lt;/strong&gt;</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 &amp;deg;C 1 hr prior surgery</td></tr><tr><td>15&quot; W x 24&quot; 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>&amp;nbsp;</td></tr><tr><td>Tape</td><td>Durapore</td><td>1538-0</td><td>&amp;nbsp;</td></tr><tr><td>1 ml Syringe</td><td>EXELint</td><td>26044</td><td>&amp;nbsp;</td></tr><tr><td>Sterile cotton tipped applicators</td><td>Kendall</td><td>8884541300</td><td>&amp;nbsp;</td></tr><tr><td>Absorbent BenchPad</td><td>VWR</td><td>56617-014</td><td>&amp;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&amp;rsquo;&amp;rsquo; x 2&amp;rsquo;&amp;rsquo; Gauze Pads</td><td>Medi-First</td><td>60673</td><td>&amp;nbsp;</td></tr><tr><td>Sterile gloves</td><td>Cardinal Health</td><td>2D7251</td><td>&amp;nbsp;</td></tr><tr><td colspan="4">&lt;strong&gt;Table 3. Equipment and instruments for I/R surgery.&lt;/strong&gt;</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

39.5K 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

Renal Ischaemia Reperfusion Injury: A Mouse Model of Injury and Regeneration

Renal ischaemia reperfusion injury (IRI) is a common cause of acute kidney injury (AKI) in patients and occlusion of renal blood flow is unavoidable during renal transplantation. Experimental models that accurately and reproducibly recapitulate renal IRI are crucial in dissecting the pathophysiology of AKI and the development of novel therapeutic agents. Presented here is a mouse model of renal IRI that results in reproducible AKI. This is achieved by a midline laparotomy approach for the surgery with one incision allowing both a right nephrectomy that provides control tissue and clamping of the left renal pedicle to induce ischaemia of the left kidney. By careful monitoring of the clamp position and body temperature during the period of ischaemia this model achieves reproducible functional and structural injury. Mice sacrificed 24 hr&#160;following surgery demonstrate loss of renal function with elevation of the serum or plasma creatinine level as well as structural kidney damage with acute tubular necrosis evident. Renal function improves and the acute tissue injury resolves during the course of 7 days following renal IRI such that this model may be used to study renal regeneration. This model of renal IRI has been utilized to study the molecular and cellular pathophysiology of AKI as well as analysis of the subsequent renal regeneration.

The mouse model of renal ischaemia reperfusion injury described here comprises of a right nephrectomy that provides control tissue and clamping of the left renal pedicle to induce ischaemia that results in acute kidney injury. This model uses a midline laparotomy approach with all steps performed via one incision.

Renal İskemi Reperfüzyon Hasarı: Yaralanma Fare Modeli ve Rejenerasyon

Renal ischaemia reperfusion injury (IRI) is a common cause of acute kidney injury (AKI) in patients and occlusion of renal blood flow is unavoidable ...

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 ...

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 ...

The Mouse Isolated Perfused Kidney Technique

The mouse isolated perfused kidney (MIPK) is a technique for keeping a mouse kidney under ex vivo conditions perfused and functional for 1 hr. This is a prerequisite for studying the physiology of the isolated organ and for many innovative applications that may be possible in the future, including perfusion decellularization for kidney bioengineering or the administration of anti-rejection or genome-editing drugs in high doses to prime the kidney for transplantation. During the time of the perfusion, the kidney can be manipulated, renal function can be assessed, and various pharmaceuticals administered. After the procedure, the kidney can be transplanted or processed for molecular biology, biochemical analysis, or microscopy.
This paper describes the perfusate and the surgical technique needed for the ex vivo perfusion of mouse kidneys. Details of the perfusion apparatus are given and data are presented showing the viability of the kidney&#39;s preparation: renal blood flow, vascular resistance, and urine data as functional, transmission electron micrographs of different nephron segments as morphological readouts, and western blots of transport proteins of different nephron segments as molecular readout.

The mouse isolated perfused kidney (MIPK) is a technique for keeping a mouse kidney under ex vivo conditions perfused and functional for 1 hr. The buffers and surgical technique are described in detail.

Fare İzole perfüze Böbrek Tekniği

The mouse isolated perfused kidney (MIPK) is a technique for keeping a mouse kidney under ex vivo conditions perfused and functional for 1 hr. This is a ...

Biyoloji

Technical Refinement of a Bilateral Renal Ischemia-Reperfusion Mouse Model for Acute Kidney Injury Research

Cardiac arrest poses a large public health burden. Acute kidney injury (AKI) is an adverse marker in survivors of cardiac arrest following the return of spontaneous circulation (ROSC) after successful cardiopulmonary resuscitation. Conversely, recovery of kidney function from AKI is a predictor of favorable neurological outcomes and hospital discharge. However, an effective intervention to prevent kidney damage caused by cardiac arrest after ROSC is lacking, suggesting that additional therapeutic strategies are required. Renal hypoperfusion and reperfusion are two pathophysiological mechanisms that cause AKI after cardiac arrest. Animal models of ischemia-reperfusion-induced AKI (IR-AKI) of both kidneys are comparable with patients with AKI following ROSC in a clinical setting. However, IR-AKI of both kidneys is technically challenging to analyze because the model is associated with high mortality and wide variation in kidney damage, which may affect the analysis. Lightweight mice were chosen, placed under general anesthesia with isoflurane, subjected to surgery with a dorsolateral approach, and their body temperature maintained during operation, thereby reducing tissue damage and establishing a reproducible acute renal IR-AKI research protocol.

This study established a protocol focusing on the technical refinement of a mouse model of bilateral renal ischemia-reperfusion for acute kidney injury research.

Akut Böbrek Hasarı Araştırmaları için Bilateral Renal İskemi-Reperfüzyon Fare Modelinin Teknik İyileştirilmesi

Cardiac arrest poses a large public health burden. Acute kidney injury (AKI) is an adverse marker in survivors of cardiac arrest following the return of ...

Developing Murine Acute Kidney Injury Model Using Bilateral Ischemia-Reperfusion

Bilateral Renal Ischemia-Reperfusion Model for Acute Kidney Injury in Mice

Acute kidney injury (AKI) is defined as a rapid decline in renal function, in which persistent kidney dysfunction gradually progresses to chronic kidney disease (CKD) due to the irreversible loss of nephrons and their maladaptive repair. In recent years, the incidence of AKI has been increasing concerning diverse etiologies, including volume depletion, sepsis, nephrotoxicity, muscle injury, and major trauma, in which ischemia-reperfusion injury (IRI)&#160;accounts for most episodes. Development of the IRI model in mice is induced by surgical clamping of the renal pedicles, which provides powerful and controllable tools for preclinical models of AKI. Importantly, the IRI model is deployed at different stages of the AKI development, especially in the processes of AKI to CKD. Despite the IRI model being widely practiced in many laboratories, a series of variables still influence the results of this model. Here, we describe the procedure of IRI model development to provide a repeatable and reliable method for researchers to explore the underlying pathogenesis in the development of AKI and the progression of AKI to CKD.

The ischemia-reperfusion injury (IRI) model can be used at different stages of acute kidney injury (AKI) development, especially during AKI to chronic kidney disease (CKD) progression. Here, we describe the procedure for IRI model development in mice via a trans-abdominal approach, clamping renal pedicles via a vascular clip to induce ischemic injury.

Acute kidney injury (AKI) is defined as a rapid decline in renal function, in which persistent kidney dysfunction gradually progresses to chronic kidney ...

Unilateral Ureteral Obstruction Model for Investigating Kidney Interstitial Fibrosis

Kidney fibrosis is the final pathological outcome of progressive chronic kidney disease (CKD). The unilateral ureteral obstruction (UUO) model is widely used to elucidate the molecular and cellular mechanisms underlying kidney interstitial fibrosis and to identify potential therapeutic targets. This model is established in mice through surgical ligation of a unilateral ureter, a procedure that is relatively straightforward and easy to perform. However, the UUO mouse model is known to exhibit significant variability and inconsistency, influenced by factors such as mouse strain, age, sex, anesthesia type, duration of surgery, body temperature during the procedure, the operator's surgical skills, feeding conditions, and the overall health status of the mice. Variations in surgical techniques, suture placement, and the duration of obstruction contribute to the variability in outcomes. Additionally, inconsistent sampling of obstructed kidneys further increases variability in the assessment of kidney fibrosis. This study outlines the process of developing the UUO mouse model and evaluating interstitial fibrosis, discusses the technical challenges contributing to the model's unpredictability, and proposes potential solutions. These insights aim to establish a more standardized and universally applicable approach for investigating kidney fibrosis.

This protocol describes a method for inducing unilateral ureteral obstruction (UUO) in mice to study the progression of tissue fibrosis in obstructive nephropathy. It includes surgical procedures, post-operative care, and methods for fibrosis assessment.

Kidney fibrosis is the final pathological outcome of progressive chronic kidney disease (CKD). The unilateral ureteral obstruction (UUO) model is widely ...