Ödülü Numaraları R01 HL064761, R25 HL059868 1SC1 HL099139 ve Sağlık P20 MD000175 formu Ulusal Sağlık Enstitüleri tarafından açıklanan proje desteklenmiştir. Içeriği sadece yazarların sorumluluğundadır ve mutlaka Ulusal Sağlık Enstitüleri resmi görüşlerini temsil etmemektedir.

1. Sıçan mezenterik arter izolasyonu <ol><li> Kapalı bir odasında izofluran ile hayvan anestezisi ve karın alkol ile silin. </li><li> Mezenterik yatakta ortaya çıkarmak için bir orta hat laparotomi gerçekleştirin. </li><li> Makaslar kullanılarak, superior mezenterik arter damarsal beslenme ile bağırsak yaklaşık 85 cm kaldırın. Pilorusta yakın bağırsak bölümünde ve bağırsak coecal kavşak yakınında distal ucu proksimal ucunu kesin. Bağırsak segmentleri açık göğüs kalp ponksiyon izofluran ve ötenazi ile derin anestezi sıçanların izole edilmiştir. </li><li> PSS içeren kaplı petri eksize bölümüne yerleştirilir ve oda sıcaklığında mezenterik arter diseksiyon. </li><li> Sağ tarafta bağırsak proksimal ucu aşağı pin ve intestineina saat yönünün tersine (bağırsak uzak tarafında sol besleme damarsal ieproximal sonuna) geri kalan pin. </li><li> Proksimal segmentinin bir parça ile birlikte şube II ve III segmentleri teşrih. </li><li> Ven inceleyin ve arter izole (V-şekilli şube noktası ile) ve yağ ve bağ dokusu ortadan kaldırarak temiz. Forseps ile yavaşça çekerek ve bağ dokusu zarından kesme arter ile doğrudan temastan kaçının. </li></ol> 2. Geminin montajı <ol><li> Ince bir forseps ile 40 mikron tungsten paslanmaz çelik tel, iki kısa kesimleri (≈ 4 cm) kesin ve endotele zarar vermemeye özen her arter lümen içine yerleştirin. Gerekirse lümen açmak için tel ucu kullanın. Kan damar dışına akışı iyi bir işaret lümen açıldı. </li><li> 115 NaCl, 4.7 KCl, 1.4 MgSO 4 .7 H 2 O, 5 NaHCO 3, 1.2 K 2 HPO 4, 1.1 Na 2 HPO 4, 1.0 CaCl 2, 20 Hepes ve: mM; fizyolojik tuz solüsyonu (PSS myograph odasının doldurun 37 5 glikoz, askorbik asit (100 mcM) ile pH 7.4) ° C ve forseps kullanarak dikkatli bir şekilde oda sıcaklığında odasına petri dişli damar segmenti transfer ve myograph odasına eksize proksimal damar segmentinin transferi ve çekme geminin içine beslemek için tel boyunca sonunda. Gemi uzanan kaçının. </li><li> Tel yakın sabitleme vidası mikrometre bağlı sağ çene altında saat yönünün tersine yakın ucunu sabitleyin. Forseps ile telin serbest ucunu yakalayın ve saat yönünde, sağ çene uzak sabitleme vidası altında güvenli. Tel boyunca geminin segment çene kendisi ile herhangi bir temas yapmadan çene arasındaki boşluğu yer olduğundan emin olun. </li><li> Dışında çene Vida ve gemi ile ikinci tel paralel hizalamak ve lümen uzak ucuna takın. Yavaşça bir kılavuz olarak önceden monte edilmiş tel kullanarak bir hareket damar segmentinin lümen yoluyla tel besleme. Gemi manevrası sırasında germe önlemek için yaklaşık 1 cm tel tutun ve endotel dokunmaktan kaçının. </li><li> Çene birlikte Vida ve ilki altındaki ikinci montaj tel hamle sağ çene güvenli olmasını sağlamak. </li><li> Yakın sabitleme vidası transdüser bağlı sol çene altında bir saat yönünde ikinci tel sona yaklaştığını sabitleyin. </li><li> Güvenli ve sabitleme vidası sol çene altında tel ucunda tel germek için sıkın. </li><li> Bir kez montajı tamamlandığında, &quot;Montaj Menü&quot; motor sıfırlamak ve geminin normalleşme başlatın. </li></ol> 3. Normalleştirme Otomatik Çift Tel Myograph System-510A &quot;Normalizasyon&quot; menüsünden değerlendirilir otomatik bir normalleştirme işlevi vardır ve gemi az 30 dakika için dengeleme sonra üretici firmanın protokolüne göre standart bir prosedür ile normalleştirilmiş bir iç çevresi gergin olması sağlar 37 ° C Üstel sonra iç çevresi basınç verilerine takılmıştır. Bu prosedür, arter, 1 100 mmHg bir transmural basınç altında rahat, in vivo olarak vardı ve var olacağını lümen çapı (d 100) tanımlar sıçan mezenterik arter için normalleşme parametreleri aşağıdaki gibidir: <ol><li> Hedef transmural basıncı = 13,3 kPa (100 mm Hg). </li><li> Zaman = 60 sn; normalleşme adımları her Süresi. </li><li> IC 1 / IC 100 = 0.9 (IC 1 = normalize iç çevresi, IC 100 = Hedef basınca tekabül eden iç çevresi . </li><li> Vizör kalibrasyon 2 * Δ (mm / oküler bölme); 2 delta programlama nedenlerden dolayı. </li></ol> Hairlines monte gemi segm uzak ve yakın uçları bittiğinde mikroskop göz parça okuma kullanarak monte mezenterik arter uzunluğunu ölçünent. Kısaca, diseksiyon mikroskobu kalibre edilmiş bir göz göz parça monte damar segmentinin uzunluğu ile maksimum büyütme ölçülür. Segmentin en uç (1) üzerinden saç hattı ile göz parça okuma ve oküler bölünmeler segmentinin yakın sonunda (2) ölçülür ve kaydedilir. Bu değerler, mikrometre okumaları, gemi germe önce ve sonra kaydedilir ve program için menüler girilen monte eğrisi ve iç çapı 100 mm Hg hedef transmural basınç karşılık gelen hesaplamak Çalışmalarda, damar lümen çapı d 1 = 0.9 etkin bir güç geliştirme maksimal xd 100,. Sıçan mezenterik arterlerde ≥ 10 mN Aktif kuvvet gelişimi devam etmek için deneyler için optimum olarak kabul edilir. Alt aktif güçle Dokular atıldı. 4. Tepkilerin ölçülmesi Normalleştirme sonra, gemilerin mekanik ve fonksiyonel özellikleri tekrarlanabilir kasılmaları elde etmek için 5 mcM fenilefrin (PE) tekrarlanan uygulamaları (normalde 2 ya da 3) ile zorlu damarları içeren bir &quot;standart start&quot; yaparak yeniden aktive edilir. Laboratuvarımızda tipik bir &quot;standart start&quot; uyarılara ve yıkanmaya dönemleri aşağıdaki gibi bir dizi oluşur: <ol><li> Myograph odası% 95 hava ve% 5 CO 2 ile kaplayın ve havalandırma değiştirin. </li><li> Taze PSS (100 mcM askorbik asit içeren) ile oda doldurun </li><li> Sözleşme 5 dakika için 5 mcM PE ile gemi ve PSS tamponu ile 4 kez yıkayın. </li><li> Sonra son yıkama, PSS ile dolum haznesi ve adımların, (ii) tekrar önce 3 dakika bekleyin ve (iii). </li></ol> Standart başladıktan sonra, arter deney için hazır. Sözleşmeli damarları gevşeten CaCl 2 artan konsantrasyonlarda (0.5 - 4 mM) kümülatif olarak değerlendirilir. inhibitörleri kullanılır. damarları deneyleri sırasında 20 dakika ve şu an için myograph odasında bileşikleri ile önceden inkübe edilir. 5. Veri Analizi SigmaPlot 11.0 grafik programı (Systat Yazılım, Richmond, CA) ile gemiler ve kuvvet iz, doğrudan metin veri dönüştürülür &quot;Normalizasyon&quot; veri analiz ve Şekil 1&#39;de gösterildiği gibi çizilir. Iz hesaplanan konsantrasyon-yanıt veriler, programın Farmakoloji menüsünde dört parametreli lojistik fonksiyonu monte deneysel veriler EC 50 değerleri ile analiz edildi. Gruplar arasında ve grup içi karşılaştırmada ise Tek Yönlü Varyans Analizi (ANOVA) ve p &lt;0.05 anlamlı olarak kabul edilir farklılıklar tarafından gerçekleştirildi. 6. Temsilcisi Sonuçlar: <table border="1"><tbody><tr><th> Parametreler </th><th> Wistar </th><th> Dahl Tuz Duyarlı </th><th> Dahl Tuz-Dirençli </th></tr><tr><td> L 100 (mikron) </td><td> 83,26 ± 2,33 </td><td> 91,52 ± 4,67 </td><td> 117,45 ± 8.43 * </td></tr><tr><td> X 1 </td><td> 604,21 ± 41,97 </td><td> 752,24 ± 85,25 </td><td> 745,84 ± 110,09 </td></tr><tr><td> r 2 </td><td> 0.99 ± 0.003 </td><td> 0.97 ± 0.008 </td><td> 0.97 ± 0.007 </td></tr></tbody></table> * Wistar ve DR L 100 değerleri arasındaki fark istatistiksel olarak anlamlı. Shapiro-Wilk Normallik Testi (p = 0,588) geçti; Eşit Varyans Testi (p = 0,237) geçti. 100 L = monte segmenti hedef transmural basınç ilgili İç çapı. X 1 = normalize iç çevresi (yani hangi deneyler yapıldı mikrometre okuma değeri) monte gemi germek için gerekli mikro pozisyoner ayarı. r 2 üstel eğriyi = oturması için regresyon katsayısı (X i, Y i). Not: Ayrıntılı prosedür 510A Myograph Sistemi 6 kılavuzda açıklanmıştır. Tablo 1: Örnek okuyun-Out &quot;Normalizasyon Menüsü&quot; nde gösterilen Basic programı Wistar, DS ve DR mezenterik arter segmentleri için normalleşme parametreleri. Değerler 5 hayvanların (± SEM) anlamına gelir. Wistar ve DS için L 100 değerleri arasındaki farklar istatistiksel olarak anlamlı (p &lt;0.05). <table border="1"><tbody><tr><th> Wistar </th><th> DS </th><th> DR </th></tr><tr><td> 14.2 ± 0.6 </td><td><strong&gt; 25.8 ± 2.7 * </td><td> 20.9 ± 1.3 * </td></tr></tbody></table> Tablo 2, her bir gemi için 5 mcM PE uygulamaları aşağıdaki, Wistar, DS ve DR sıçan izole mezenterik arterler, geliştirilen gerilimler ( MN). Bildirilen değerlerin 6-8 hayvanların (± SEM) anlamına gelir. * Wistar denetimleri önemli derecede farklılık (p &lt;0.05). <img alt="Şekil 1" src="/files/ftp_upload/3119/3119fig1.jpg" /> Şekil 1. Ca 2 + PE-sözleşmeli bir Wistar sıçan mezenterik arter e-kaynaklı rahatlama fizyolojik tuz çözeltisi içeren 1 mM Ca 2 + tel myograph monte edilmiş . Arter 30 dakika süreyle 37 ° dengelenmiş ° C sürekli havalandırma ve yanıtları Ca 2 + e belirlenir. PE Ca 2 kümülatif eklemeler 5 mcM ek dayanan bir temsilci force + gösterilmiştir. <img alt="Şekil 2" src="/files/ftp_upload/3119/3119fig2.jpg" /> Şekil 2 Ca 2 + PE-sözleşmeli DS sıçan mezenterik arter e-kaynaklı rahatlama fizyolojik tuz çözeltisi içeren 1 mM Ca 2 + tel myograph monte edilmiş . Arter 30 dakika süreyle 37 ° dengelenmiş ° C sürekli havalandırma ve yanıtları Ca 2 + e belirlenir. PE Ca 2 kümülatif eklemeler 5 mcM ek dayanan bir temsilci force + gösterilmiştir. Ca 2 + rahatlama ciddi Wistar ve DR sıçan göre bu doku içinde tatlıya bağlandı. <img alt="Şekil 3" src="/files/ftp_upload/3119/3119fig3.jpg" /> Şekil 3. Ca 2 + PE-DR sıçan sözleşmeli mezenterik arter e-kaynaklı rahatlama fizyolojik tuz çözeltisi içeren 1 mM Ca 2 + tel myograph monte edilmiş . Arter dengelenmiş az 30 dakika süreyle 37 ° C sürekli havalandırma ve yanıtları Ca 2 + 5 mcM PE Ca 2 kümülatif eklenen ek iz e determined.A temsilcisi force + gösterilmiştir. <img alt="Şekil 4" src="/files/ftp_upload/3119/3119fig4.jpg" /> Şekil 4. A. [Ca 2 +] E-yanıt eğrileri B. DR ve DS sıçanların izole mezenter eğlencelerle dört parametre lojistik curve.C.CaR ifade veri sıkıştırılmak suretiyle belirlenir dinlenmek için EC 50 değerlerini gösteren Çubuk grafik.

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Çıkar çatışması ilan etti.

Hipertansiyon, kalp-damar, beyin ve böbrek morbidite / mortalitenin önde gelen nedenidir. Hipertansiyon oluşumu nüfusun yüksek olduğu ve tuz duyarlı hipertansiyon özellikle yaşlanan nüfus, yüksek ve siyahlar beyazlara nazaran daha yaygındır. Bunun nedeni, onların böbrekler 9 sodyum korumak için siyahların eğilimi olduğuna inanılıyor . Tuz duyarlılık, böbrek hastalığı için önemli bir faktör ve endotel disfonksiyonu ile ilişkili, fakat mekanizması tam olarak anlaşılmış değil...

<table border="1"><tbody><tr><th> Ekipman / reaktif Adı </th><th> Şirket </th><th> Katalog numarası </th><th> Yorumlar (isteğe bağlı) </th></tr><tr><td> Otomatik Çift Tel Myograph System-510A DMT-USA, Inc. </td><td> Atlanta, GA. </td><td> 100151 </td><td></td></tr><tr><td> PowerLab/4SP Veri Toplama Sistemi </td><td> ADInstruments, Colorado Springs, CO </td><td> ML750 </td><td> 4-16 giriş kanalları ile yeni modeller mevcuttur. </td></tr><tr><td> Dell Dimension XPS Gen 4 Bilgisayar </td><td> Vadi </td><td></td><td></td></tr><tr><td> STEMI SV II (Apo) Oküler ile Diseksiyon Mikroskop </td><td> Carl Zeiss Uluslararası. </td><td></td><td></td></tr><tr><td> Wistar, Dahl tuz duyarlı, Dahl tuz dirençli sıçanların </td><td> IN Harlan Sprague Dawley, Indianapolis,. </td><td></td><td></td></tr><tr><td> Kemirgen chow </td><td> Harland Tekland, Madison, WI. </td><td></td><td></td></tr><tr><td> Fenilefrin </td><td> Sigma-Aldrich, St Louis, MO. </td><td></td><td></td></tr><tr><td> Diğer kimyasallar </td><td></td><td></td><td> Kullanılan tüm kimyasallar, piyasada bulunan en saf notları. </td></tr></tbody></table>

mesenteric artery contraction and relaxation studies using automated wire myography

Proksimal direnç damarları, mezenterik arterler gibi periferik direnç önemli ölçüde katkıda bulunur. 100-400 mm arasında çapa işlevi öncelikle vücudun genel gereksinimlerine göre çeşitli organlara kan akışını yönlendirerek bu küçük gemiler. Sıçan mezenterik arter, 100 mm daha büyük bir çapa sahip. Kasılmaların kâğıda çizilmesi tekniği, ilk olarak Mulvay ve Halpern 1 açıklanan Bevan ve Osher 2 tarafından önerilen yöntem dayanmaktadır . Teknik kas hazırlanmasında önemli bir kısalma önlenir izometrik koşullar altında, küçük gemiler hakkında bilgi sağlar. Farklı agonistleri gemilerin yürürlüğe üretim ve duyarlılık aktif gerilim uzunlukta ilişkisi göre, streç ölçüde bağımlı olduğu için, montaj teller uygun önlemek için izometrik koşullar altında daralma çalışmalar yapmak şarttır. Paslanmaz çelik teller kaydedilen yanıtlar 3 etkileyen ikincisi, oksidasyonu nedeniyle tungsten teller tercih edilir. Tekniği, damar düz kas hücre reseptörleri normal fonksiyonu için kanıt elde etmek için monte gemilerin agonist bağlı kasılmalar karşılaştırılmasına olanak sağlar . Biz phenylyephrine ile sözleşmeli izole mezenterik arterler ekstrasellüler kalsiyum (Ca 2 + e) kümülatif konsantrasyonlarda eklenmesi üzerine dinlenmek çeşitli çalışmalarda gösterilmiştir. Bulgular bize G-protein-birleştiğinde Ca 2 +-algılayan reseptör (SYR) ifade eden bu perivasküler duyusal sinirler, sonucuna götürdü, bu vasorelaxation yanıt arabuluculuk . Otomatik bir tel kasılmaların kâğıda çizilmesi yöntemi kullanarak, Wistar, Dahl tuz-duyarlı (DS) ve tuza dayanıklı Dahl (DR) mezenterik arterler sıçan Ca 2 + e farklı yanıt verdiğini göstermektedir. Wistar sıçanlarda Dokular + hassasiyet DR ve DS olanlara kıyasla daha yüksek Ca 2 gösterdi. DS sıçan mezenterik arterler azalma araba ifade azalır Ca 2 + e-indüklenen izole, ön sözleşmeli arterlerin rahatlama ile ilişkilidir. Veri, araba, mezenterik arterler artmış adrenerjik ton altında rahatlamak için gerekli olduğunu öne sürdüğü gibi hipertansiyon oluşur ve çok daha şiddetli DS Dahl hayvanlarda araba sinyal yolağı içsel bir kusur, göstermektedir. Yöntem, mezenterik direnç arter ve farklı agonist ve / veya antagonistleri arasındaki benzer küçük kan damarlarını ve karşılaştırmalar vasküler reaktivite ex vivo belirlenmesinde yararlı yan yana 6,7,8 kolay ve sürekli olarak değerlendirilir olabilir .

Watch this Scientific Journal Video about Mesenteric Artery Contraction and Relaxation Studies Using Automated Wire Myography at JoVE.com

Mesenteric Artery Contraction and Relaxation Studies Using Automated Wire Myography

Izole mezenterik arterler kuvvet ölçümleri için otomatik bir kasılmaların kâğıda çizilmesi yöntemi açıklanmıştır. Fenilefrin ve ekstrasellüler kalsiyum yanıtlarını belirlemek için 510A Mulvany Halpern Otomatik Çift Tel Myograph kullanır. 300 mikron, bağımsız yöntemi 60 çaplarının küçük damarlarda agonistleri izometrik yanıtları tutarlı tespitine imkan sağlamaktadır.

Otomatik Tel kasılmaların kâğıda çizilmesi Yüklü daralma ve Rahatlama Çalışmaları

Proximal resistance vessels, such as the mesenteric arteries, contribute substantially to the peripheral resistance. These small vessels of between ...

mesenteric-artery-contraction-relaxation-studies-using-automated-wire

Research

JoVE Journal

Medicine

28.4K Görüntüleme.  North Carolina Central University, Durham. Izole mezenterik arterler kuvvet ölçümleri için otomatik bir kasılmaların kâğıda çizilmesi yöntemi açıklanmıştır. Fenilefrin ve ekstrasellüler kalsiyum yanıtlarını belirlemek için 510A Mulvany Halpern Otomatik Çift Tel Myograph kullanır. 300 mikron, bağımsız yöntemi 60 çaplarının küçük damarlarda agonistleri izometrik yanıtları tutarlı tespitine imkan sağlamaktadır.

Video: Mesenteric Artery Contraction and Relaxation Studies Using Automated Wire Myography

Assessing Myogenic Response and Vasoactivity In Resistance Mesenteric Arteries Using Pressure Myography

Small resistance arteries constrict and dilate respectively in response to increased or decreased intraluminal pressure; this phenomenon known as myogenic response is a key regulator of local blood flow. In isobaric conditions small resistance arteries develop sustained constriction known as myogenic tone (MT), which is a major determinant of systemic vascular resistance (SVR). Hence, ex vivo&#160;pressurized preparations of small resistance arteries are major tools to study microvascular function in near-physiological states. To achieve this, a freshly isolated intact segment of a small resistance artery (diameter ~260 &#956;m) is mounted onto two small glass cannulas and pressurized. These arterial preparations retain most in vivo characteristics and permit assessment of vascular tone in real-time. Here we provide a detailed protocol for assessing vasoactivity in pressurized small resistance mesenteric arteries from rats; these arteries develop sustained vasoconstriction - approximately 25% of maximal diameter - when pressurized at 70 mmHg. These arterial preparations may be used to study the effect of investigational compounds on relationship between intra-arterial pressure and vasoactivity and determine changes in microvascular function in animal models of various diseases.

Pressure myography is used to assess vasoactivity of small arteries that develop sustained constriction when pressurized. This manuscript provides a detailed protocol to assess in isolated segments of small mesenteric arteries from rats, vasoactivity and the effect of intraluminal pressure on vascular diameter.

Basınç myography kullanma Direnç Mezenterik Arter ise andMyogenic Tepkisi ve vazoaktiviteye Değerlendirilmesi

Small resistance arteries constrict and dilate respectively in response to increased or decreased intraluminal pressure; this phenomenon known as myogenic ...

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Isolating and Using Sections of Bovine Mesenteric Artery and Vein as a Bioassay to Test for Vasoactivity in the Small Intestine

Mammalian gastrointestinal systems are constantly exposed to compounds (desirable and undesirable) that can have an effect on blood flow to and from that system. Changes in blood flow to the small intestine can result in effects on the absorptive functions of the organ. Particular interest in toxins liberated from feedstuffs through fermentative and digestive processes has developed in ruminants as an area where productive efficiencies could be improved. The video associated with this article describes an in vitro bioassay developed to screen compounds for vasoactivity in isolated cross-sections of bovine mesenteric artery and vein using a multimyograph. Once the blood vessels are mounted and equilibrated in the myograph, the bioassay itself can be used: as a screening tool to evaluate the contractile response or vasoactivity of compounds of interest; determine the presence of receptor types by pharmacologically targeting receptors with specific agonists; determine the role of a receptor with the presence of one or more antagonists; or determine potential interactions of compounds of interest with antagonists. Through all of this, data are collected real-time, tissue collected from a single animal can be exposed to a large number of different experimental treatments (an in vitro advantage), and represents vasculature on either side of the capillary bed to provide an accurate picture of what could be happening in the afferent and efferent blood supply supporting the small intestine.

The small intestine is frequently exposed to toxins that can influence blood flow and negatively impact nutrient absorption. Using a multimyograph and mesenteric artery and vein isolates, compounds or toxins of interest can be screened for vasoactivity.

Bir Biyoassay olarak Sığır Mezenterik Arter ve Ven Bölüm yalıtma ve kullanma İnce Barsak vazoaktiviteye için Test

Mammalian gastrointestinal systems are constantly exposed to compounds (desirable and undesirable) that can have an effect on blood flow to and from that ...

Murine Model of Femoral Artery Wire Injury with Implantation of a Perivascular Drug Delivery Patch

Percutaneous interventions including balloon angioplasty and stenting have been used to restore blood flow in vessels with occlusive vascular disease. While these therapies lead to the rapid restoration of blood flow, these technologies remain limited by restenosis in the case of bare metal stents and angioplasty, or reduced healing and possibly enhanced risk of thrombosis in the case of drug eluting stents. A key pathophysiological mechanism in the formation of restenosis is intimal hyperplasia caused by the activation of vascular smooth muscle cells and inflammation due to arterial stretch and injury. Surgeries that induce arterial injury in genetically modified mice are useful for the mechanistic study of the vascular response to injury but are often technically challenging to perform in mouse models due to the their small size and lack of appropriate sized devices. We describe two approaches for a surgical technique that induces endothelial denudation and arterial stretch in the femoral artery of mice to produce robust neointimal hyperplasia. The first approach creates an arteriotomy in the muscular branch of the femoral artery to obtain vascular access. Following wire injury this arterial branch is ligated to close the arteriotomy. A second approach creates an arteriotomy in the main femoral artery that is later closed through localized cautery. This method allows for vascular access through a larger vessel and, consequently, provides a less technically demanding procedure that can be used in smaller mice. Following either method of arterial injury, a degradable drug delivery patch can be placed over or around the injured artery to deliver therapeutic agents.

We describe a surgical technique that produces wire injury in the femoral artery of mice to induce neointimal hyperplasia to serve as a model testing system for the perivascular delivery of therapeutic compounds for the inhibition of restenosis.

Bir Perivasküler Drug Delivery Patch İmplantasyonunda ile Femoral Arter Tel Yaralanma sıçan Modeli

Percutaneous interventions including balloon angioplasty and stenting have been used to restore blood flow in vessels with occlusive vascular disease. ...

A Murine Model of Arterial Restenosis: Technical Aspects of Femoral Wire Injury

Cardiovascular disease caused by atherosclerosis is the leading cause of death in the developed world. Narrowing of the vessel lumen, due to atherosclerotic plaque development or the rupturing of established plaques, interrupts normal blood flow leading to various morbidities such as myocardial infarction and stroke. In the clinic endovascular procedures such as angioplasty are commonly performed to reopen the lumen. However, these treatments inevitably damage the vessel wall as well as the vascular endothelium, triggering an excessive healing response and the development of a neointimal plaque that extends into the lumen causing vessel restenosis (re-narrowing). Restenosis remains a major cause of failure of endovascular treatments for atherosclerosis. Thus, preclinical animal models of restenosis are vitally important for investigating the pathophysiological mechanisms as well as translational approaches to vascular interventions. Among several murine experimental models, femoral artery wire injury is widely accepted as the most suitable for studies of post-angioplasty restenosis because it closely resembles the angioplasty procedure that injures both endothelium and vessel wall. However, many researchers have difficulty utilizing this model due to its high degree of technical difficulty. This is primarily because a metal wire needs to be inserted into the femoral artery, which is approximately three times thinner than the wire, to generate sufficient injury to induce prominent neointima. Here, we describe the essential surgical details to effectively overcome the major technical difficulties of this model. By following the presented procedures, performing the mouse femoral artery wire injury becomes easier. Once familiarized, the whole procedure can be completed within 20 min.

The mouse femoral artery wire injury model of restenosis is technically challenging. In this protocol we show the key technical details essential for successfully performing wire injury to induce consistent&#160;neointima for studies of restenosis.

Arter Restenozun bir fare modeli: Femur Tel Yaralanma Teknik Yönleri

Cardiovascular disease caused by atherosclerosis is the leading cause of death in the developed world. Narrowing of the vessel lumen, due to ...

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. ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

İzometrik Contractility ölçüm tel Myography kullanarak fare mezenterik arter

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

Model of Ischemic Heart Disease and Video-Based Comparison of Cardiomyocyte Contraction Using hiPSC-Derived Cardiomyocytes

Ischemic heart disease is a significant cause of death worldwide. It has therefore been the subject of a tremendous amount of research, often with small-animal models such as rodents. However, the physiology of the human heart differs significantly from that of the rodent heart, underscoring the need for clinically relevant models to study heart disease. Here, we present a protocol to model ischemic heart disease using cardiomyocytes differentiated from human induced pluripotent stem cells (hiPS-CMs) and to quantify the damage and functional impairment of the ischemic cardiomyocytes. Exposure to 2% oxygen without glucose and serum increases the percentage of injured cells, which is indicated by staining of the nucleus with propidium iodide, and decreases cellular viability. These conditions also decrease the contractility of hiPS-CMs as confirmed by displacement vector field analysis of microscopic video images. This protocol may furthermore provide a convenient method for personalized drug screening by facilitating the use of hiPS cells from individual patients. Therefore, this model of ischemic heart disease, based on iPS-CMs of human origin, can provide a useful platform for drug screening and further research on ischemic heart disease.

We present a model of ischemic heart disease using cardiomyocytes derived from human induced pluripotent stem cells, together with a method for quantitative evaluation of tissue damage caused by ischemia. This model can provide a useful platform for drug screening and further research on ischemic heart disease.

HiPSC Türetilmiş Kardiyomiyosit Kullanarak İskemik Kalp Hastalığı Modeli ve Kardiyomiyosit Kontraksiyonunun Video Tabanlı Karşılaştırılması

Ischemic heart disease is a significant cause of death worldwide. It has therefore been the subject of a tremendous amount of research, often with ...

Mechanical Control of Relaxation Using Intact Cardiac Trabeculae

Diastolic dysfunction is a common phenotype across cardiovascular disease presentations. In addition to elevated cardiac stiffness (elevated left ventricular end-diastolic pressure), impaired cardiac relaxation is a key diagnostic indicator of diastolic dysfunction. While relaxation requires the removal of cytosolic calcium and deactivation of sarcomeric thin filaments, targeting such mechanisms has yet to provide effective treatments. Mechanical mechanisms, such as blood pressure (i.e., afterload), have been theorized to modify relaxation. Recently, we showed that modifying the strain rate of a stretch, not afterload, was both necessary and sufficient to modify the subsequent relaxation rate of myocardial tissue. The strain rate dependence of relaxation, called the mechanical control of relaxation (MCR), can be assessed using intact cardiac trabeculae. This protocol describes the preparation of a small animal model, experimental system and chamber, isolation of the heart and subsequent isolation of a trabecula, preparation of the experimental chamber, and experimental and analysis protocols. Evidence for lengthening strains in the intact heart suggests that MCR might provide new arenas for better characterization of pharmacological treatments, along with a method to assess myofilament kinetics in intact muscles. Therefore, studying the MCR may elucidate a path to novel approaches and new frontiers in the treatment of heart failure.

Rapid myocardial and cardiac relaxation is essential for normal physiology. Mechanical relaxation mechanisms are now known to be dependent on strain rate. This protocol provides an overview of the acquisition and analysis of experiments to further study the mechanical control of relaxation.

Sağlam Kardiyak Trabeküller Kullanılarak Gevşemenin Mekanik Kontrolü

Diastolic dysfunction is a common phenotype across cardiovascular disease presentations. In addition to elevated cardiac stiffness (elevated left ...

Multimodality Diagnosis of Mesenteric Ischemia

Early diagnosis of mesenteric ischemia remains challenging because mesenteric ischemia presents with no key symptoms or physical findings, and no laboratory data specifically indicates intestinal tissue ischemic status before necrosis develops. While computed tomography is the standard for diagnostic imaging, there are several limitations: (1) repeated assessments are associated with increased radiation exposure and risk of renal damage; (2) the computed tomography findings can be misleading because necrosis occasionally occurs despite opacified mesenteric arteries; and (3) computed tomography is not necessarily available within the golden time of salvaging the intestines for those patients in the operating room or at a place far from the hospital. This article describes a challenge to overcome such limitations using ultrasonography and near-infrared light, including clinical studies. The former is capable of providing not only morphologic and kinetic information of the intestines but also perfusion of the mesenteric vessels in real-time without transferring the patient or exposing them to radiation. Transesophageal echocardiography enables precise assessment of mesenteric perfusion in the OR, ER, or ICU. Representative findings of mesenteric ischemia in seven aortic dissection cases are presented. Near-infrared imaging with indocyanine green helps visualize the perfusion of vessels and intestinal tissues although this application requires laparotomy. Findings in two cases (aortic aneurysm) are shown. Near-infrared spectroscopy demonstrates oxygen debt in the intestinal tissue as digital data and can be a candidate for early detection of mesenteric ischemia without laparotomy. The accuracy of these assessments has been confirmed by intraoperative inspections and postoperative course (prognosis).

This article presents a multimodal approach that aims to overcome the limitations of traditional methods in detecting mesenteric ischemia and preventing bowel necrosis. The presented technique offers a promising solution by combining state-of-the-art ultrasonography with cutting-edge near-infrared light technologies.

Mezenterik İskeminin Multimodalite Tanısı

Early diagnosis of mesenteric ischemia remains challenging because mesenteric ischemia presents with no key symptoms or physical findings, and no ...

Rodent Model of Intestinal Ischemia-Reperfusion Injury via Occlusion of the Superior Mesenteric Artery

Intestinal ischemia-reperfusion injury (IRI) is associated with a myriad of conditions in both veterinary and human medicine. Intestinal IRI conditions, such as gastric dilatation volvulus (GDV), mesenteric torsion, and colic, are observed in animals such as dogs and horses. An initial interruption of blood flow causes tissues to become ischemic. Although necessary to salvage viable tissue, subsequent reperfusion can induce further injury. The main mechanism responsible for IRI is free radical formation upon reperfusion and reintroduction of oxygen into damaged tissue, but there are many other components involved. The resulting local and systemic effects often impart a poor prognosis. 
Intestinal IRI has been the subject of extensive research over the past 50 years. An in vivo rodent model in which the base of the superior mesenteric artery (SMA) is temporarily ligated is currently the most common method used to study intestinal IRI. Here, we describe a model of intestinal IRI utilizing isoflurane anesthesia in 21% O2 medical air that yields reproducible injury, as demonstrated by consistent histopathology of the small intestines. Tissue injury was also assessed in the colon, liver, and kidneys.

We describe how to generate a widely used surgical model of intestinal ischemia-reperfusion injury (IRI) in rodents. The procedure involves occlusion of the superior mesenteric artery followed by the restoration of blood flow. This model is useful for studies investigating occlusive causes of intestinal IRI in both veterinary and human medicine.

Superior mezenterik arterin tıkanması yoluyla intestinal iskemi-reperfüzyon hasarının kemirgen modeli

Intestinal ischemia-reperfusion injury (IRI) is associated with a myriad of conditions in both veterinary and human medicine. Intestinal IRI conditions, ...