Name: a rat orthotopic renal transplantation model for renal allograft rejection
Uploaded: 2022-02-02T15:00:00
Description: Watch this Scientific Journal Video about A Rat Orthotopic Renal Transplantation Model for Renal Allograft Rejection at JoVE.com

这项工作得到了中国国家自然科学基金（81870304）对李俊的支持，以及Else Kröner-Fresenius-Stiftung（Nr. 2017_A28）对Marcus Groettrup的支持。

商业上获得的近交8-10周龄雄性F344和Lewis大鼠（200g至250g）。同种异体左肾移植在雄性F344和Lewis大鼠之间进行。F344大鼠被用作供体和同源受体，Lewis大鼠用作同种异体受体。所有动物处理程序均按照NIH发布的《实验动物护理和使用指南》进行，所有动物实验方案均获得重庆大学肿瘤医院动物护理使用委员会的批准。手术期间使用的所有用品，包括手术器械和溶液，都是无菌的。该协议的原理图如图 ...

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大鼠肾移植是一项具有挑战性的工作，需要高水平的显微手术技术，并且操作技术已经过多次优化。从一开始，Gonzalez等人将供体肾脏植入受体的颈部，并将供体输尿管吻合到皮肤9。然而，由于泌尿管感染和供体输尿管狭窄的高发生率，该手术在短时间内被放弃。随后，通过供体肾动脉和肾静脉与受体主动脉和下腔静脉15的吻合，或通过供体肾动脉和肾静脉与?...

肾移植已成为终末期肾功能衰竭患者最有效的治疗方法。然而，T细胞介导的急性排斥反应和同种抗体介导的体液免疫排斥反应导致肾同种异体移植物的病理性损伤，并限制了肾移植后患者的短期和长期生存1，2，3。不幸的是，仍然缺乏防止肾脏同种异体移植物排斥反应的有效药物，因为肾同种异体移植物的免疫和病理排斥反应的确切机制尚不清楚。因此，阐明肾同种异体移植物的免疫和病理排斥机制的临床前研究有助于寻找新的靶点并开发相关的有效药物，以防止肾脏同种异体移植物的排斥反应并最终延长患者的生存期。
最近在原位肾移植的大鼠模型研究中提出了肾同种异体移植排斥反应的许多潜在免疫学和病理生理学机制4，5，6，7，8。这些发现提出了几种新的靶点和相关干扰方法，作为抑制肾脏同种异体排斥反应的有希望的治疗方法，例如补体调节因子和抗CD59抗体6，免疫蛋白酶体和环氧酮抑制剂7，8。因此，大鼠原位肾移植是研究肾移植后肾同种体移植物免疫排斥和病理损伤机制的理想临床前模型。
大鼠肾移植已逐渐从供体肾脏9 的异位植入转向使用血管的端到端吻合术或使用袖带方法的输尿管的端到端吻合术的原位肾植入10，11，12。本研究描述了大鼠间对受者肾动脉和肾静脉进行端到端吻合术的大鼠正位肾移植的详细方案，以及输尿管-膀胱吻合术的端到端“隧道”方法，避免了下半身缺血和下腔静脉血栓形成，减少了术后漏尿和输尿管扭曲。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>&#160;10-0 Polyamide Monofilament suture</td>
			<td>B.Braun Medical Inc.</td>
			<td>G0090781</td>
			<td></td>
		</tr>
		<tr>
			<td>&#160;4-0 Polyamide Monofilament suture</td>
			<td>B.Braun Medical Inc.</td>
			<td>C1048451</td>
			<td></td>
		</tr>
		<tr>
			<td>&#160;8-0 Polyamide Monofilament suture</td>
			<td>B.Braun Medical Inc.</td>
			<td>C2090880</td>
			<td></td>
		</tr>
		<tr>
			<td>Buprenorphine</td>
			<td>US Biological life Sciences</td>
			<td>352004</td>
			<td></td>
		</tr>
		<tr>
			<td>Electrocoagulator</td>
			<td>Electrocoagulator</td>
			<td>ZJ1099</td>
			<td></td>
		</tr>
		<tr>
			<td>F344 and Lewis rats</td>
			<td>Center of Experimental Animals (Tongji Medical College, Huazhong University of Science and Technology, China)</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr>
			<td>Gauze</td>
			<td>Henan piaoan group Co., LTD</td>
			<td>10210402</td>
			<td></td>
		</tr>
		<tr>
			<td>Heating pad</td>
			<td>Guangzhou Dewei Biological Technology Co., LTD</td>
			<td>DK0032</td>
			<td></td>
		</tr>
		<tr>
			<td>Heparin</td>
			<td>North China Pharmaceutical Co., LTD</td>
			<td>2101131-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Injection syringe (1 ml and 10 ml)</td>
			<td>Shandong weigao group medical polymer Co., LTD</td>
			<td>20211001</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>RWD Life Science Co., LTD</td>
			<td>21070201</td>
			<td></td>
		</tr>
		<tr>
			<td>Penicillin G Sodium</td>
			<td>Wuhan HongDe Yuexin pharmatech co.,Ltd</td>
			<td>69-57-8</td>
			<td></td>
		</tr>
		<tr>
			<td>Scalp needle (24 G)</td>
			<td>Hongyu Medical Group</td>
			<td>20183150210</td>
			<td></td>
		</tr>
		<tr>
			<td>Shaver</td>
			<td>Beyotime</td>
			<td>FS600</td>
			<td></td>
		</tr>
		<tr>
			<td>Small animal anesthesia machine</td>
			<td>RWD Life Science</td>
			<td>R500</td>
			<td></td>
		</tr>
		<tr>
			<td>Small Animal Surgery Kit</td>
			<td>Beyotime</td>
			<td>FS500</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium chloride injection</td>
			<td>Southwest pharmaceutical Co., LTD</td>
			<td>H50021610</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical operation microscope</td>
			<td>Tiannuoxiang Scientific Instrument Co. , Ltd, Beijing, China</td>
			<td>SZX-6745</td>
			<td></td>
		</tr>
		<tr>
			<td>Swab</td>
			<td>Yubei Medical Materials Co., LTD</td>
			<td>21080274</td>
			<td></td>
		</tr>
		<tr>
			<td>Tape</td>
			<td>Minnesota Mining Manufacturing Medical Equipment (Shanghai) Co., LTD</td>
			<td>1911N68</td>
			<td></td>
		</tr>
		<tr>
			<td>UW solution</td>
			<td>Bristol-Myers Squibb Company</td>
			<td>17HB0002</td>
			<td></td>
		</tr>
	</tbody>
</table>

a rat orthotopic renal transplantation model for renal allograft rejection

肾同种异体移植排斥反应限制了肾移植后患者的长期生存。大鼠原位肾移植是临床前研究中研究肾同种异体移植排斥机制的基本模型，有助于开发提高肾同种异体移植物长期生存率的新方法。大鼠原位肾移植中的供体肾植入通常通过对受者的主动脉和下腔静脉进行端到端吻合。在该模型中，使用端到端吻合口将供体的肾脏植入接受者的肾动脉和肾静脉。供体的输尿管以端到端的“隧道”方法吻合受者的膀胱。该模型有助于更好地愈合输尿管 - 膀胱吻合口，并通过避免干扰血液供应和下半身静脉反流来提高接受者的生存率。该模型可用于研究急性和慢性免疫和肾脏同种异体移植物的病理排斥的机制。在这里，该研究描述了大鼠之间这种原位肾移植的详细方案。

在该大鼠原位肾移植模型中，受体大鼠在手术后正常移动。为了观察肾同种异体移植的慢性排斥反应，移植后将受体大鼠饲养10周，此时受者大鼠的总存活率约为90%。死亡的主要原因是术后出血和漏尿。其他主要并发症包括手术期间出血、肾血管血栓形成和肾积水，其发病率分别约为 15%、25% 和 20%。手术过程中的大部分出血可以停止，并且不影响受体大鼠的存活。应从后续检查中排除血管栓塞和?...

Watch this Scientific Journal Video about A Rat Orthotopic Renal Transplantation Model for Renal Allograft Rejection at JoVE.com

A Rat Orthotopic Renal Transplantation Model for Renal Allograft Rejection

大鼠原位肾移植模型有助于研究肾同种异体移植排斥的机制。目前的模型使用肾脏植入的端到端吻合术和输尿管 - 膀胱吻合术的端到端“隧道”方法，在不干扰血液供应和下半身静脉反流的情况下增加了受者的生存率。

一种用于肾同种异体排斥的大鼠原位肾移植模型

Renal allograft rejection limits the long-term survival of patients after renal transplantation. Rat orthotopic renal transplantation is an essential ...

大鼠原位肾移植是研究临床前研究中肾同种异体移植排斥机制的重要模型，有助于开发提高肾同种异体移植物长期生存率的新方法。该手术中肾血管的端到端吻合术避免了下半身缺血和下腔静脉血栓形成，而在端到端隧道方法中输尿管吻合术可减少术后尿漏和输尿管扭曲。该方法为研究肾移植后肾同种异体移植物免疫排斥和损伤的机制提供了见解。首先使用手术剪刀沿着从耻骨联合到尾下腓的中线切开腹部。然后使用牵开器拉开切口。从切口右侧取出肠道，并用湿润的纱布包裹它们，以防止它们变干。然后暴露左肾。使用微镊子和棉签在20倍放大镜下解离左肾动脉和静脉。如有必要，对肾上腺静脉和生殖器静脉进行缝合和横切。在微镊子的帮助下解离输尿管，保留外周结缔组织，以确保输尿管的血液供应。使用血管钳或 4-0 聚酰胺单丝缝合线，分离并夹住或连接左肾动脉上方约 5 毫米的主动脉。然后将左肾静脉远端横切至左生殖器静脉和肾上腺静脉的结合处。使用24号手术刀针头，用冰冷的UW溶液从左肾动脉下方的主动脉冲洗肾脏，并补充每毫升100单位肝素，直到血液褪色。切除主动脉旁边的左肾动脉，然后切除膀胱旁边的输尿管，并在微镊子和剪刀的帮助下解离肾脏。将供体左肾保存在冰冷的UW溶液中。在20倍放大下，使用棉签和微镊子解离受体大鼠的左肾动脉和肾静脉。如有必要，调整肾上腺静脉和生殖器静脉。使用微血管钳夹夹左肾动脉和肾静脉，然后在距离微血管钳夹三毫米的地方横切肾动脉和肾静脉。解离并连接左侧输尿管，在肾脏下方约两到三厘米处，8-0聚酰胺单丝缝合并在连接处将其横切。通过在微镊子和剪刀的帮助下解离肾脏来切除接受者天然左肾。将供体肾脏植入受体大鼠的左肾窝中，并在植入的供体肾脏周围放置冰块。切换到45倍放大，然后使用10-0聚酰胺单丝缝合线以端到端模式将供体肾动脉和肾静脉吻合到受者肾动脉和肾静脉。要使用中断缝线吻合肾动脉，请将停留缝合线放在吻合口的12点和6点钟位置。用两到三针在两条缝线之间的吻合口的一侧等距缝合。翻转留置缝合线，同样用两到三针缝合两个留置缝线之间的吻合口的另一侧缝合。要使用连续缝线吻合肾静脉，请将停留缝合线放在吻合口的6点和12点钟位置。使用连续缝线从12点钟位置缝合吻合口的一侧，缝制四到五针。翻转留置缝合线，同样，从六点钟位置缝合吻合口的另一侧。通过释放肾静脉的非侵入性微血管钳来重新灌注供体肾脏，识别出血部位并在必要时进行额外的缝合。接下来，释放肾动脉的非侵入性微血管钳，识别出血部位，并在必要时进行额外的缝合。在20倍放大下，用4-0聚酰胺单丝缝合线作为牵引线连接供体输尿管的末端，以将末端拖过受者膀胱的隧道。通过将供体输尿管的外膜与受者膀胱的肌肉层缝合在四个等距位置，使用8-0将供体输尿管与受者膀胱固定在一起聚酰胺单丝缝合线。之后，用受者膀胱外的缝合线切除供体输尿管的末端，并允许供体输尿管收缩回受者膀胱。将肠道放回腹腔，并在肌肉层中连续缝合缝合腹部切口，然后用4-0聚酰胺单丝缝合线关闭皮肤层。移植后十周，苏木精和曙红染色以及周期性移酸染色显示肾小球硬化症，间质纤维化，肾同种异体移植物中的肾小管萎缩和间质性动脉硬化症，与等移植肾形成鲜明对比。银染色显示同种异体移植肾与异位移植物肾相比，同种异体肾小球基底膜增厚，表明大鼠原位肾移植模型的成功。由于肾脏的血管孔很薄，为了避免错误的缝合，记得在高倍率场上进行吻合。

Research

JoVE Journal

Medicine

Orthotopic Aortic Transplantation: A Rat Model to Study the Development of Chronic Vasculopathy

主动脉原位移植大鼠模型，研究慢性血管病变的发展

Research models of chronic rejection are essential to investigate pathobiological and pathophysiological processes during the development of transplant ...

科研

医学

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

大鼠肾移植后急性排斥反应的非侵入性的成像使用<sup&gt; 18</sup&gt; F-FDG PET

The number of patients with end-stage renal  disease, and the number of kidney allograft recipients continuously increases.  Episodes of acute cellular ...

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

部分原位肝移植大鼠模型的肝动脉重建的外科手术

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

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

肾缺血再灌注损伤：损伤的小鼠模型和再生

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

Mouse Kidney Transplantation: Models of Allograft Rejection

鼠肾移植：移植排斥反应模型

Rejection of the transplanted kidney in humans is still a major cause of morbidity and mortality. The mouse model of renal transplantation closely ...

Murine Kidney Transplant Technique

小鼠肾脏移植技术

The first mouse kidney transplant technique was published in 19731 by the Russell laboratory. Although it took some years for other labs to become ...

Heterotopic Renal Autotransplantation in a Porcine Model: A Step-by-Step Protocol

异位自体肾移植的猪模型：有步骤，分步协议

Kidney transplantation is the treatment of choice for patients suffering from end-stage renal disease. It offers better life expectancy and higher quality ...

A Rat Model of Orthotopic Liver Transplantation Using a Novel Magnetic Anastomosis Technique for Suprahepatic Vena Cava Reconstruction

应用新型磁吻合技术 Suprahepatic 腔静脉重建大鼠原位肝移植模型的研究

The rat model of orthotopic liver transplantation (OLT) is essential for transplant research. It is a very sophisticated animal model and requires a steep ...

An Efficient Sieving Method to Isolate Intact Glomeruli from Adult Rat Kidney

一种从成年大鼠肾脏中分离完整型肾小球的有效筛分方法

Preservation of glomerular structure and function is pivotal in the prevention of glomerulonephritis, a category of kidney disease characterized by ...

Incorporation of a Survivable Liver Biopsy Procedure in Mice to Assess Non-alcoholic Steatohepatitis (NASH) Resolution

Incorporation of a Survivable Liver Biopsy Procedure in Mice to Assess Non-alcoholic Steatohepatitis NASH Resolution

在小鼠中纳入可存活的肝脏活检程序评估非酒精性脂肪性肝炎 (NASH) 的解决方法

Clinical trials assessing therapies for the treatment of non-alcoholic steatohepatitis (NASH) involve a baseline and end of study liver biopsy, and ...