Name: shunt surgery, right heart catheterization, and vascular morphometry in a rat model for flow-induced pulmonary arterial hypertension
Uploaded: 2017-02-11T13:00:00
Description: Watch this Scientific Journal Video about Shunt Surgery, Right Heart Catheterization, and Vascular Morphometry in a Rat Model for Flow-induced Pulmonary Arterial Hypertension at JoVE.com

This study was supported by the Netherlands Cardiovascular Research Initiative, the Dutch Heart Foundation, the Dutch Federation of University Medical Centers, the Netherlands Organization for Health Research and Development, and the Royal Netherlands Academy of Sciences (CVON nr. 2012-08, PHAEDRA, The Sebald fund, Stichting Hartekind).

動物を対象とする手順は、動物実験のためにオランダ中央委員会と大学医療センターフローニンゲンでの動物保護委員会（NL）によって承認されています。 180〜300 gの量の両方のWistar及びルイスラットを使用しました。 1.住宅と順化<ol><li>ケージ当たり5のグループ内の中央動物施設、家のラットに到着した後。 7日間の順応期間中は、人間の取り扱いにラットを慣らす?...

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この方法は、流れ誘発性PAHおよびPAHと、このモデルを特徴付ける原則血行動態及び組織病理学的エンドポイントを評価するためのテクニックを作成するために、MCTで前処置したラットにおける大動脈大静脈シャントの外科的手順を説明します。 プロトコルおよびトラブルシューティングの中で重要なステップ手術と術後。大動脈大静脈シャント手術中?...

この方法の目的は、ラットにおける重度、流れによって誘発される肺動脈高血圧症の再現可能なモデルを作成するために、その原理血行動態及び組織病理学的エンドポイントを測定することです。 肺動脈性肺高血圧症（PAH）、右心室不全や死に至る肺血管抵抗の漸進的増加を含む臨床症候群です。肺高血圧疾患（PH）の上位疾患スペクトラムの中で、PAHは治癒1せずに残っている最も深刻なフォームと1です。 PAHにおける基礎となる動脈症は、血管内腔を閉塞血管リモデリングの典型的な形が特徴です。通常の非muscularized血管のMuscularizationと内側容器層の肥大はPAHの初期疾患現象とみなされ、また、PH 2の他の形態で見られる、と3可逆的であると考えられています。 PAH Aとしてdvances、内膜層は、最終的に特徴的な新生内膜病変2を形成し 、改造を開始します。新生内膜型肺血管リモデリングは、PAHに排他的であり、現在4不可逆的であるとみなされています。 PAHは稀な疾患であるため、その病理生物学的理解の進歩と新しい治療法の開発は、動物モデルに大きく依存してきました。ラットにおけるmonocrotalin（MCT）モデルがされている単純な単一のヒットモデルであり、まだ、頻繁に使用されます。 MCTは、肺動脈と地域炎症5への損傷を引き起こす毒素です。 4週6から60ミリグラム/キロのMCTは、3後の平均肺動脈圧（MPAP）の増加、肺血管抵抗（PVR）、および右心室肥大（RVH）につながります。組織形態は、新生内膜病変5せずに孤立した内側の肥大によって特徴付けられます。 MCTそれは一般的に、後者のように示されるが、ラットモデルは、このように、適度なPHの形ではなく、PAHを表します。 先天性左から右へのシャント（PAH-CHD）に関連付けられているPAHの小児では、肺の血流が新生内膜病変7、8、9の開発に不可欠なトリガとしてみなされている増加しました。ラットでは、腹部大動脈及び大静脈、第1990 10に記載された技術との間のシャントを作成することによって誘導することができ、肺の血流を増加させました。増加した肺の流れを作成するための選択肢が一方的摘術により、または肺動脈吻合部11に鎖骨によるものです。これらのモデルの概念の欠点は、または起因する肺血管系の医原性損傷の肺切除によって誘発され、残りの肺および適応経路活性化の潜在的な代償成長から成り肺動脈吻合に、両方の増加した肺の血流の影響を混乱させる。 大動脈大静脈シャントが作成され、MCT投与ラットにおける第二のヒットとして誘発される肺の血流を増加させると、特徴的な新生内膜病変が発生し、増加した後にPAHの重症型と関連した右心室不全（RVF）は、3週間の開発します12を流れます。このモデルにおけるPAHの血行力学的進行は心エコー検査および右心カテーテル法によってインビボで評価することができます。血管の組織形態学、血管壁の厚さ、動脈閉塞の程度、および右心室不全のためのパラメータは、PAHのex vivo特徴付けの柱を形成します。 この方法は、大動脈大静脈シャント（AC-シャント）手術、右心カテーテル法、および血管組織形態の定性的および定量的評価のための詳細なプロトコルを記述しています。

<table border="0" cellpadding="0" cellspacing="0" width="835">
	<tbody>
		<tr height="25">
			<td height="25" style="height:25px;width:307px;">
			Shunt Surgery
			</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td style="width:196px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Sterile surgical gloves</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Duratears Eye ointment</td>
			<td style="width:151px;">Alcon</td>
			<td style="width:183px;">10380</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Chloride-Hexidine</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Cotton swabs</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Histoacryllic tissue glue</td>
			<td style="width:151px;">B. Braun Medical</td>
			<td>1050052</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Silkam 5-0 sutures black non-resorbable</td>
			<td style="width:151px;">B. Braun Medical</td>
			<td>F1134027</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Safil 4-0 sutures violet resorbable</td>
			<td style="width:151px;">B. Braun Medical</td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">18 G needle&#160;</td>
			<td style="width:151px;">Luer</td>
			<td>NN1838R BD</td>
			<td>tip bent in 45 degrees orifice to the outside</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Gauzes 10x10 cm</td>
			<td>Paul Hartmann</td>
			<td>407825</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Temgesic Buprenorphine</td>
			<td style="width:151px;">RB Pharmaceuticals</td>
			<td style="width:183px;">5429</td>
			<td>subcutaneous injection</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Sodium Chloride 0.9 %</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Ventilation mask Rat</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Scalple blade</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Biemer clamp 18 mm, 5 mm opening&#160;</td>
			<td><a href="https://www.google.nl/url?sa=i&amp;rct=j&amp;q=&amp;esrc=s&amp;source=images&amp;cd=&amp;cad=rja&amp;uact=8&amp;ved=0ahUKEwitqICpkITNAhWiL8AKHe6iCC8QjhwIBQ&amp;url=http%3A%2F%2Fwww.agnthos.se%2Fquote%2Fquote%2Fcaturl%2Fid%2F242%2FBiemer&amp;psig=AFQjCNEnfRXuMQ6Doi4_-2KMYu3qeRovMA&amp;ust=1464777707579746">AgnTho</a></td>
			<td>64-562</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Heat mat</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Kocher Clamp</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Shaving machine</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Microscope</td>
			<td style="width:151px;">Leica</td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:307px;">
			Right Heart Catheterization
			</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Name</td>
			<td style="width:151px;">Company</td>
			<td style="width:183px;">Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Sterile surgical gloves</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Eye ointment</td>
			<td style="width:151px;">Duratears</td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Chloride-Hexidine</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Cotton swabs</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Gauzes 10x10 cm</td>
			<td>Paul Hartmann</td>
			<td>407825</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Silkam 5-0 sutures black non-resorbable</td>
			<td style="width:151px;">B. Braun Medical</td>
			<td>F1134027</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Needle 20 G</td>
			<td style="width:151px;">Luer</td>
			<td style="width:183px;"></td>
			<td>Tip slightly bent to the inside</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Cannula 20 G</td>
			<td style="width:151px;">Luer</td>
			<td style="width:183px;"></td>
			<td>to introduce catheter, tip pre-formed in 20 degrees</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Silastic Catheter 15 cm long</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td>0.5 mm ball 2 mm from tip</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Pressure transducer</td>
			<td style="width:151px;">Ailtech</td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Bedside monitor Cardiocap/5</td>
			<td style="width:151px;">Datex-Ohmeda</td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Shaving machine</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">10mL Syringe</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Sodium Chloride 0.9 %</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td>for flushing</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:307px;">
			Vascular Morphology
			</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Name</td>
			<td style="width:151px;">Company</td>
			<td style="width:183px;">Catalog Number</td>
			<td>Comments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">50ml Syringe</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">4 % Formaldehyde</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">18 G cannula with tube</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Verhoef staining kit</td>
			<td style="width:151px;">Sigma-Aldrich</td>
			<td><a href="http://www.sigmaaldrich.com/catalog/product/sigma/ht254?lang=en&amp;region=NL">HT254</a></td>
			<td>http://www.sigmaaldrich.com/catalog/product/sigma/ht254?lang=en&#38;region=US</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Digital slide scanner</td>
			<td style="width:151px;">Hamamatsu</td>
			<td style="width:183px;">C9600</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Image-J</td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Elastic (Connective Tissue Stain)&#160;</td>
			<td style="width:151px;">Abcam</td>
			<td style="width:183px;">ab150667</td>
			<td>http://www.abcam.com/elastic-connective-tissue-stain-ab150667.html</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;"></td>
			<td style="width:151px;"></td>
			<td style="width:183px;"></td>
			<td>http://www.abcam.com/ps/products/150/ab150667/documents/ab150667-Elastic%20Stain%20Kit%20(website).pdf</td>
		</tr>
	</tbody>
</table>

shunt surgery, right heart catheterization, and vascular morphometry in a rat model for flow-induced pulmonary arterial hypertension

In this protocol, PAH is induced by combining a 60 mg/kg monocrotalin (MCT) injection with increased pulmonary blood flow through an aorto-caval shunt (MCT+Flow). The shunt is created by inserting an 18-G needle from the abdominal aorta into the adjacent caval vein. Increased pulmonary flow has been demonstrated as an essential trigger for a severe form of PAH with distinct phases of disease progression, characterized by early medial hypertrophy followed by neointimal lesions and the progressive occlusion of the small pulmonary vessels. To measure the right heart and pulmonary hemodynamics in this model, right heart catheterization is performed by inserting a rigid cannula containing a flexible ball-tip catheter via the right jugular vein into the right ventricle. The catheter is then advanced into the main and the more distal pulmonary arteries. The histopathology of the pulmonary vasculature is assessed qualitatively, by scoring the pre- and intra-acinar vessels on the degree of muscularization and the presence of a neointima, and quantitatively, by measuring the wall thickness, the wall-lumen ratios, and the occlusion score.

代表的な結果を図4に示されています。コントロール（N = 3）、MF8（N = 5）、MF14（N = 5）、MF28（N = 5）、およびMF-RVF（：示された結果は、以下の群内のLewisラットにおけるMCT + FLOWの特性を示しますN = 10）。統計分析は、ボンフェローニ補正を用いて、一方向ANOVAを用いて行きました。 60ミリグラム/キログラムMCT?...

Watch this Scientific Journal Video about Shunt Surgery, Right Heart Catheterization, and Vascular Morphometry in a Rat Model for Flow-induced Pulmonary Arterial Hypertension at JoVE.com

Shunt Surgery, Right Heart Catheterization, and Vascular Morphometry in a Rat Model for Flow-induced Pulmonary Arterial Hypertension

This protocol describes a surgical procedure to create a model for flow-induced pulmonary arterial hypertension (PAH) in rats and the procedures to analyze the principle hemodynamic and histological end-points in this model.

流れ誘導性の肺動脈性肺高血圧症のためのラットモデルにおけるシャント手術、右心カテーテル、および血管形態計測

In this protocol, PAH is induced by combining a 60 mg/kg monocrotalin (MCT) injection with increased pulmonary blood flow through an aorto-caval shunt ...

Research

JoVE Journal

Medicine

Femoral Arterial and Venous Catheterization for Blood Sampling, Drug Administration and Conscious Blood Pressure and Heart Rate Measurements

血液サンプリング、医薬品の管理と意識血圧および心拍数測定のための大腿動脈および静脈カテーテル

In multiple fields of study, access to the circulatory system in laboratory studies is necessary. Pharmacological studies in rats using chronically ...

Surgical Procedures for a Rat Model of Partial Orthotopic Liver Transplantation with Hepatic Arterial Reconstruction

肝動脈再建を用いた部分同所性肝移植のラットモデルのための外科的処置

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as ...

Echocardiographic Assessment of the Right Heart in Mice

マウスの右心の心エコー評価

Transgenic and toxic models of pulmonary arterial hypertension (PAH) are widely used to study the pathophysiology of PAH and to investigate potential ...

Chronic Thromboembolic Pulmonary Hypertension and Assessment of Right Ventricular Function in the Piglet

慢性血栓塞栓性肺高血圧症とピグレットにおける右室機能の評価

An original piglet model of Chronic Thromboembolic Pulmonary Hypertension (CTEPH) associated with chronic Right Ventricular (RV) dysfunction is described. ...

Hemodynamic Characterization of Rodent Models of Pulmonary Arterial Hypertension

肺動脈性肺高血圧症の齧歯類モデルの血行動態特性評価

Pulmonary arterial hypertension (PAH) is a rare disease of the pulmonary vasculature characterized by endothelial cell apoptosis, smooth muscle ...

Induction and Phenotyping of Acute Right Heart Failure in a Large Animal Model of Chronic Thromboembolic Pulmonary Hypertension

慢性血栓塞栓性肺高血圧症の大型動物モデルにおける急性右心不全の誘導とフェノタイピング

The development of acute right heart failure (ARHF) in the context of chronic pulmonary hypertension (PH) is associated with poor short-term outcomes. The ...

Left Atrial Stenosis Induced Pulmonary Venous Arterialization and Group 2 Pulmonary Hypertension in Rat

左心房狭窄による肺静脈動脈化術とグループ 2 肺高血圧症ラット

The mechanism of mitral stenosis-induced pulmonary venous arterialization and group 2 pulmonary hypertension (PH) is unclear. There is no rodent model of ...

Invasive Hemodynamic Assessment for the Right Ventricular System and Hypoxia-Induced Pulmonary Arterial Hypertension in Mice

マウスにおける右心室系と低酸素誘発性肺動脈高血圧に対する侵襲的血行力学的評価

Pulmonary arterial hypertension (PAH) is a chronic and severe cardiopulmonary disorder. Mice are a popular animal model used to mimic this disease. ...

A Rat Carotid Artery Pressure-Controlled Segmental Balloon Injury with Periadventitial Therapeutic Application

過期治療アプリケーションを用いてラット頸動脈圧制御区分バルーン損傷

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