Name: establishment and evaluation of a porcine vein graft disease model
Uploaded: 2022-07-25T14:00:00
Description: Watch this Scientific Journal Video about Establishment and Evaluation of a Porcine Vein Graft Disease Model at JoVE.com

作者感谢广东省实验动物监测研究所的技术支持、动物护理和样本采集。他们还感谢深圳市迈瑞生物医疗电子有限公司在超声检测方面的技术支持。这项工作得到了中国广东省科技计划和暨南大学中央大学基础科研业务费用项目（2017A020215076、2008A08003和21621409）的支持。

实验动物的护理和使用程序经广东省实验动物监测研究所机构动物护理和使用委员会批准。所有实验均按照《实验动物护理和使用指南》（2011年第8版，美国国家研究委员会）进行。手术过程如图 1所示。
1.动物术前准备
<ol><li>将10头体重30-35公斤的3个月大的雄性迷你猪随机分为假猪组（n = 5）和VGD组（n = 5）。</li>
	<li>使用体重指数（BMI）?...

<ol>
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在这项研究中，我们详细描述了在开发CABG诱导的VGD模型时动物选择，器械准备，外科手术和术后评估的方案。我们在CABG手术前后对静脉移植物进行了超声检查，并在手术后30天对移植物进行了组织学检查。在冠状动脉旁路移植术之前，乳内静脉的血流是正常的，而在乳内静脉的移植物中观察到逆行血流。与假手术组相比，手术组动物的肝肾功能受到一定程度的损害。考虑到冠状动脉移植疾病的发...

尽管近年来冠心病死亡率显著下降，但美国每年有一半的中年人出现缺血性心脏相关症状，三分之一的老年人死于冠心病1。冠状动脉旁路移植术（CABG）是改善心肌缺血的有效手术方式，更重要的是，它是治疗多血管冠状动脉疾病不可替代的手术方式2。然而，随着时间的推移，血管移植物会出现炎症、内膜增生和进行性动脉粥样硬化，已知会导致静脉移植失败或静脉移植疾病 （VGD）3。在冠状动脉旁路移植术后的患者中，如果发生再狭窄，在某些情况下只能更换患病的血管2。老年患者和增加的合并症使得重做冠状动脉旁路移植术非常具有挑战性。延缓或控制与移植血管相关的病理问题是一个亟待解决的问题。需要CABG-VGD的大型动物模型来研究疾病机制和制定治疗策略。研究人员已经成功地在小鼠4，大鼠5，兔子6和猪7等小型和大型动物中建立了动物VGD模型。与小动物相比，猪等大型动物具有与人类相似的解剖结构和生理特征，寿命更长8，9。因此，大型动物更适合于探索静脉移植疾病的长期病理变化和药物或器械的临床前测试。我们和我们的合作团队已经成功地应用手术技术建立了猪心力衰竭模型，并描述了该模型中的心脏病理变化10。
CABG手术在临床实践中已经标准化，但当它应用于VGD动物模型的建立时，物种之间的差异，动物设备和设施的获取，动物外科手术以及动物饲养和护理是研究人员面临的巨大挑战。与临床实践一样，用于建立VGD动物模型的CABG手术方法包括中线胸骨切开术11 和左外侧开胸术12。中线胸骨切开术更常用13，14。然而，这种方法对人类和动物都有很高的风险。在Thankam等人报告的研究中，用于建模的六头猪中有两头在手术中死亡15。高模型死亡率会增加研究成本并影响结果的准确性。早些时候的一项研究表明，在猪中，左胸壁切口可以建立CABG诱导的VGD11。在这里，本研究旨在描述一种循序渐进的方案，为小型猪中CABG诱导的VGD模型建立可重复的手术，并评估该模型的表型。实验方案由心脏外科和麻醉团队共同设计。手术前根据实验室中其他迷你猪的尸体确定左第三肋间隙的手术入路，并根据中心16使用的方法进行麻醉方法。通过血液生化检查、超声检查和组织学检查来评估动物模型。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Aortic Punch</td>
			<td>Medtronic Inc. , America</td>
			<td>3.0mm, 3.5mm, 4.0mm</td>
			<td>Used for proximal coronary bridge anastomosis</td>
		</tr>
		<tr>
			<td>Automatic biochemical analyzer</td>
			<td>IDEXX Laboratories, Inc. America</td>
			<td>Catalyst One</td>
			<td></td>
		</tr>
		<tr>
			<td>Cardiac coronary artery bypass grafting instrument kit</td>
			<td>LANDANGER, France</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cardiogram monitor</td>
			<td>Shenzhen Mindray Bio-Medical Electronics Co, Ltd</td>
			<td>MEC-1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Coronary Shunt</td>
			<td>AXIUS&#160;</td>
			<td>OF-1500, OF-2500, OF-3000</td>
			<td>The product temporarily blocks the coronary artery during arteriotomy to reduce the amount of bleeding in the surgical field and provide blood flow to the distal end during anastomosis.&#160; The Axius shunt plug is not an implant and should be removed prior to completion of the anastomosis.&#160;&#160;</td>
		</tr>
		<tr>
			<td>Defibrillator</td>
			<td>MEDIANA</td>
			<td>Mediana D500</td>
			<td></td>
		</tr>
		<tr>
			<td>Diazepam</td>
			<td>Nanguo pharmaceutical Co. LTD, Guangdong, China</td>
			<td>H37023039&#160;</td>
			<td>Narcotic inducer</td>
		</tr>
		<tr>
			<td>Disposable manual electric knife</td>
			<td>Covidien, America</td>
			<td>E2516H</td>
			<td></td>
		</tr>
		<tr>
			<td>Electric negative pressure suction machine</td>
			<td>Shanghai Baojia Medical Instrument Co, Ltd</td>
			<td>YX932D</td>
			<td></td>
		</tr>
		<tr>
			<td>Esmolol</td>
			<td>Guangzhou Wanzheng Pharmaceutical Co. LTD</td>
			<td>H20055990</td>
			<td>Emergency drugs</td>
		</tr>
		<tr>
			<td>Ice machine&#160;</td>
			<td>Local suppliers, Guangzhou, China</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Lidocaine&#160;</td>
			<td>Chengdu First Pharmaceutical Co. LTD</td>
			<td>H51021662</td>
			<td>Emergency drugs</td>
		</tr>
		<tr>
			<td>Luxtec headlight system</td>
			<td>Luxtec, America</td>
			<td>AX-1375-BIF</td>
			<td>Used for lighting fine parts during operation</td>
		</tr>
		<tr>
			<td>Medical operation magnifier (glasses)</td>
			<td>Germany Lista co, LTD</td>
			<td>SuperVu Galilean 3.5&#215;</td>
			<td>Used for fine site operation during operation</td>
		</tr>
		<tr>
			<td>Multi-function high-frequency electrotome</td>
			<td>Shanghai Hutong Electronics Co, Ltd</td>
			<td>GD350-B</td>
			<td></td>
		</tr>
		<tr>
			<td>Nitrogen canister</td>
			<td>Local suppliers, Guangzhou, China</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Nonabsorbable surgical suture (polypropylene suture)</td>
			<td>Johnson &#38; Johnson, America</td>
			<td>6-0, 7-0</td>
			<td>Used to suture blood vessels.</td>
		</tr>
		<tr>
			<td>Nonabsorbable suture (cotton thread)</td>
			<td>Covidien, America</td>
			<td>1-0</td>
			<td>Used for skin and muscle tissue tugging</td>
		</tr>
		<tr>
			<td>Open heart surgery instrument kit</td>
			<td>Shanghai Medical Instrument (Group) Co., LTD</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Propofol injection</td>
			<td>Xi &#39;an Libang Pharmaceutical Co. LTD</td>
			<td>H19990282</td>
			<td>Anesthetic sedative</td>
		</tr>
		<tr>
			<td>Refrigerator</td>
			<td>Local suppliers, Guangzhou, China</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Respiratory anesthesia machine for animal</td>
			<td>Shenzhen Reward Life Technology Co, Ltd, China</td>
			<td>R620-S1</td>
			<td></td>
		</tr>
		<tr>
			<td>Semi-occlusion clamp</td>
			<td>Xinhua Surgical Instrument Co., Ltd.</td>
			<td>ZL1701RB</td>
			<td>Temporarily cut off the aortic flow</td>
		</tr>
		<tr>
			<td>vecuronium bromide</td>
			<td>Richter, Hungary&#160;</td>
			<td>JX20090127</td>
			<td>Muscle relaxant</td>
		</tr>
		<tr>
			<td>Veterinary ultrasound system&#160;</td>
			<td>Royal Philips, Netherlands</td>
			<td>CX50</td>
			<td></td>
		</tr>
		<tr>
			<td>Zoletil</td>
			<td>Virbac, France</td>
			<td>Zoletil 50&#160;</td>
			<td>Animal narcotic</td>
		</tr>
	</tbody>
</table>

establishment and evaluation of a porcine vein graft disease model

静脉移植疾病 （VGD） 是冠状动脉旁路移植术 （CABG） 失败的主要原因。需要CABG-VGD的大型动物模型来研究疾病机制和制定治疗策略。 
为了进行手术，我们通过第三肋间隙进入心腔，仔细解剖乳腺内静脉并将其浸入生理盐水中。然后治疗右主冠状动脉缺血。切开目标血管，放置分流栓，并吻合移植静脉远端。升主动脉部分阻塞，穿孔后吻合移植静脉近端。检查移植静脉是否通畅，并结扎右冠状动脉近端。 
在迷你猪中进行冠状动脉旁路移植术，以收获左内乳静脉以用作血管移植物。血清生化检查用于评估手术后动物的生理状态。超声检查显示移植血管近端、中端和远端无梗阻。在手术模型中，在CABG手术后的组织学检查中观察到移植物中的湍流血流，并且在移植物中观察到与内膜增生相关的静脉移植物狭窄。本研究为建立可重复的冠状动脉旁路移植术诱导的VGD模型提供了详细的外科手术。

BMI 和血清生化指标 假和VGD组之间的BMI没有显着差异（假与VGD，22.05公斤/厘米2 ±0.46公斤/厘米2 对21.14公斤/厘米2 ±0.39公斤/厘米2， p = 0.46）。血清生化结果列于 表1。4个生化指标显示组间差异有统计学意义，包括天冬氨酸氨基转移酶（AST，假与VGD，25.25 IU/L±1.88 IU/L±31.5 IU/L2.58 IU/L），血清胆红素（假胆红素与VGD，2.5μmol/L±0.47μmol/L?...

Watch this Scientific Journal Video about Establishment and Evaluation of a Porcine Vein Graft Disease Model at JoVE.com

Establishment and Evaluation of a Porcine Vein Graft Disease Model

在该方案中，通过左胸壁上的小切口进行新型猪静脉旁路移植术，而无需体外循环。进行了术后病理学研究，显示内膜增厚。

猪静脉移植病模型的建立与评价

Venous graft disease (VGD) is the leading cause of coronary artery bypass graft (CABG) failure. Large animal models of CABG-VGD are needed for the ...

该模型为建立冠状动脉移植疾病的动物模型提供了标准方案。该方法建立的动物模型程序是标准的，结果稳定。首先，将三个月大的雄性迷你猪体重30至35公斤分为假和静脉移植疾病组，每组五只动物。麻醉动物后，通过将留置静脉导管插入边缘耳静脉来建立耳通路。现在，将猪以仰卧位转移到手术台上。使用 7.0 至 7.5 法式规格插管进行气管插管，并将其连接到麻醉呼吸回路。用绷带固定四肢，并用无菌窗帘抬高头部。静脉注射维库溴铵以松弛肌肉后，使用心电图监测心率、血氧水平和体温。剃须并消毒手术区域后，在其周围放置无菌手术单。如文所述切开并露出乳内静脉后，将乳内静脉和左侧乳内动脉定位在胸骨左侧，并用血管钳对乳内静脉进行钝性解剖。通过用电刀电凝左内乳静脉分支进行止血。如果止血不完全，使用棉线，在收获时结扎并标记静脉的两端。一旦静脉被移除，将肝素，生理盐水注射到静脉中进行预处理。然后，将静脉放入生理盐水中，保留备用，并同样切开假组的乳腺内静脉。然后，打开心包，闭合胸壁，使用乳内静脉进行病理控制，无需冠状动脉旁路移植。用电刀在心包上做一个大约七厘米的切口，露出右冠状动脉干。暂停心包，并用1-0手术缝合线缝合同侧的皮肤。现在，将右冠状动脉干与周围组织分开。接下来，使用线钩绕过主动脉附近孤立的右冠状动脉近端下方的阻断带。然后，通过收紧和放松阻断带，用两个分钟缺血和五个分钟再灌注的三个周期治疗心肌。此外，在缺血再灌注预处理期间用心电图监测心脏的电活动。接下来，在收紧束带后切开覆盖血管的心外膜，以阻断右冠状动脉血流。暴露冠状动脉壁，用手术刀片的尖端纵向切开血管前壁的中心。切开管腔并扩大切口并放置冠状动脉分流后，用线圈将分流的一端通过撕裂插入远端冠状动脉。现在，将冠状动脉中的血液分流到空心冠状动脉分流器中，以确保手术区域畅通，并使用6-0聚丙烯缝合线在乳腺内静脉和主动脉壁之间进行端到端连续缝合。在升主动脉的中间，用半闭塞夹住升主动脉的左前外侧壁。在切开外膜的主动脉壁上用手术刀片做一个小切口后，通过这个切口将冲头头端的滑动轴的头端插入主动脉腔。然后，向外收缩滑动轴，圆刀切断一块动脉壁。一旦分流器被拉出，用 6-0 聚丙烯缝合线在乳内静脉和右冠状动脉之间进行端到端连续缝合。打开半闭塞夹后，使用超声记录右冠状动脉干近端吻合部位的旁路流量。如前所述，为动物准备手术。并用电刀在手术后 30 天用电刀做一个 10 厘米的胸骨正中切口以分割胸骨并收获静脉。分离时，避开主要血管、心脏，将移植的血管逐层分离。迅速切断连接心脏的大血管，将心脏和升主动脉放在冰片上。一旦移植血管桥、连接的主动脉和右冠状动脉被移除，用生理盐水在 4 摄氏度下冲洗所有样本。取整个移植容器约三到四厘米并将其分成四到五个相等的部分后，将其转移到冷冻管中，在液氮中快速快速冷冻。为了进行分析，将冰冷的盐水冲洗的移植物固定在4%多聚甲醛溶液中。组织固定12小时后，将切片染色在50毫升苏木精水溶液中三分钟。然后，用50毫升0.5%盐酸-乙醇和0.2%氨水洗涤各10秒钟，将切片分开。用流水冲洗切片一小时后，将它们浸泡在蒸馏水中三分钟来清洁它们。在70%和90%乙醇中脱水10分钟后，将它们放入50毫升0.5%酒精曙红染色溶液中2至3分钟。一旦染色部分在纯乙醇中脱水，将样品浸泡在纯二甲苯中10分钟，使其透明。最后，用中性胶水滴下透明部分，然后用盖玻片覆盖它们。在光学显微镜下以40倍放大倍率观察病理切片。超声检查显示，移植血管近端、血管腔、远端整体血流方向正常。然而，移植血管的近端和远端显示出一些逆行的血液流动。乳内静脉的组织学分析显示，假组的被膜内膜、被膜中层和静脉移植物的静脉壁正常。然而，冠状动脉移植后30天，血管内膜增厚，管腔变窄。假移植物和静脉移植物病组中，天冬氨酸氨基转移酶、血清胆红素、总胆红素、肌酐4项生化指标变化有统计学意义。外科医生应完全暴露乳腺内部静脉，并确保它没有附着在周围组织上。该技术为研究动物模型中的静脉移植疾病提供了标准化的建模过程。

Research

JoVE Journal

Medicine

A Multicenter MRI Protocol for the Evaluation and Quantification of Deep Vein Thrombosis

深静脉血栓形成的评价和定量的多中心MRI协议

We evaluated a magnetic resonance venography (MRV) approach with gadofosveset to quantify total thrombus volume changes as the principal criterion for ...

科研

医学

Establishment and Characterization of UTI and CAUTI in a Mouse Model

建立和UTI和CAUTI表征在小鼠模型

Urinary tract infections (UTI) are highly prevalent, a significant cause of morbidity and are increasingly resistant to treatment with antibiotics. ...

Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency

静脉介入型号：研究旁路移植通畅一个合适的模型

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

用线 Myography 测定小鼠肠系膜动脉的等轴收缩力

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

Establishment of a Mouse Severe Acute Pancreatitis Model using Retrograde Injection of Sodium Taurocholate into the Biliopancreatic Duct

建立小鼠重症急性胰腺炎模型，将牛磺胆酸钠逆行注射到胆胰管中

The prevalence of acute pancreatitis (AP), especially severe acute pancreatitis (SAP), is increasing in younger age groups annually. However, there is a ...

Repetitive Blood Sampling from the Subclavian Vein of Conscious Rat

Rats are widely used in pharmacokinetics (PK) and toxicokinetic (TK) studies that need to collect a certain amount of blood at specific time points to ...

Modified Arteriovenous Fistula Surgery to Establish the RV Volume Overload Mouse Model

Establishment and Confirmation of a Postnatal Right Ventricular Volume Overload Mouse Model

Establishment and Confirmation of a Postnatal Right Ventricular Volume Overload Mouse Model (Video) | JoVE 

出生后右心室容积超负荷小鼠模型的建立与确认

Right ventricular (RV) volume overload (VO) is common in children with congenital heart disease. In view of distinct developmental stages,the RV ...

Comparative Study of Blood Collection Techniques in Rats

Modified Tail Vein and Penile Vein Puncture for Blood Sampling in the Rat Model

Context Dependent Blood Collection Methods in Rats (Video) | JoVE

改良尾静脉和静脉穿刺用于大鼠模型中的血液采样

Blood samples are required in most experimental animal designs to assess various hematological parameters. This paper presents two procedures for blood ...

Adjunctive Exosome Therapy: Enhancing Myocardial Recovery Post-CABG Surgery

Surgical Porcine Model of Chronic Myocardial Ischemia Treated by Exosome-laden Collagen Patch and Off-pump Coronary Artery Bypass Graft

Author Spotlight: Enhancing Coronary Artery Revascularization

Chronic myocardial ischemia resulting from progressive coronary artery stenosis leads to hibernating myocardium (HIB), defined as myocardium that adapts ...

Development of an Effective Mice Model for Studying Oronasal Fistulas

Establishment of an Oronasal Fistula Mice Model

Investigating Wound Healing in Mice Models of Oronasal Fistulas (Video) | JoVE

口鼻瘘小鼠模型的建立

This study presents a method utilizing heated ophthalmologic cautery to develop a viable model for investigating oronasal fistulas. C57BL/6 mice were used ...