这项研究得到了德国研究基金会 (DFG, AR 1092/2-1) 的资助。

所有的方法都是按照国家卫生研究院的指导, 对实验动物的护理和使用以及德国的动物福利法进行的, 并得到地方当局 (Regierungspräsidium Tübingen) 的批准。
1. 阿霉素注射液球窦诱导实验性肾病综合征
<ol><li>通过标记活塞的停止位置, 准备一个装有30G 套管的0.5 毫升注射器。</li>
	<li>计算男性阿霉素的注射量 (7.25 µL/克体重 (体重) 等于14.5 µg/克阿霉素) 和雌性小鼠 (6.9 µL/克体重等于13.8 µg/克阿霉素) 根据从早上的体重。 注: 给定剂量适用于肾病综合征的诱导。剂量的细微变化导致了一个关键的不同病程的肾病从轻度慢性肾脏疾病, 只有小的变化, 肾小球滤过率 (12.6 µg/克阿霉素) 急性肾衰竭 (15.4 µg/g 生物武器阿霉素)7, 15。</li>
	<li>在37摄氏度温暖的房间里加热计算出的阿霉素溶液 (2 µg/µL) 的数量。 注意: 阿霉素是有害的, 如果吞咽和可能导致癌症。如果与阿霉素合作, 避免皮肤接触, 一定要戴上手套。</li>
	<li>预先填充与阿霉素溶液的注射器, 直到标记点, 并设置平衡为零。用注射量填充注射器, 通过称量注射器检查体积。</li>
	<li>Narcotize 用5体积% 异氟醚与2升/分钟的气化器和氧气流进行深度的小鼠。<ol>
<li>注射前不要用眼膏。操作者需要清楚地看到轨道腔, 以执行安全和成功的球注射。</li>
		<li>在开始注射前, 通过踏板反射评估麻醉的水平并调整麻醉分娩。</li>
	</ol>
</li>
	<li>将鼠标放在右侧侧卧床, 其背部面向操作员的身体, 其头部面向操作员的注射手。 注: 该程序被描述为一个右手的人, 为左手的人执行注射与左手, 它是更容易做的位置和注射反向右向左。</li>
	<li>通过对皮肤, 背部和腹侧的眼睛, 小心地将鼠标左眼球从眼窝上伸出。</li>
	<li>穿刺左球窦从内眼角度 (内侧眼睑合生面)。避免与 mouse´s 眼球接触。</li>
	<li>稍微倾斜注射器和注入整个体积一去。确保注入的体积没有任何阻力, 没有任何渗出迹象, 如 exopthalmus 或渗漏从注射现场。</li>
	<li>由于阿霉素是一种剧毒物质, 每天至少检查一次, 使用一个评分表根据莫顿和格里菲斯et等。16 , 并特别注意注入站点。如果任何迹象如 exopthalmus, 眼睑关闭或任何坏死迹象如肿胀或皮肤病变发生注射后或在接下来的日子里, 老鼠应该被安乐死。</li>
</ol>2. 含有抑肽酶的缓释微丸的植入
<ol><li>用每天需要的剂量和提前释放时间来订购颗粒。在抑肽酶的情况下, 选择每天1毫克的剂量, 在10天内释放。 注: 在本研究中, 抑肽酶颗粒含有10毫克的剂量。<ol>
<li>在干燥的条件下储存小球, 避免接触湿气。</li>
	</ol>
</li>
	<li>为手术准备必需的物品, 包括一把剪刀, 一双皮肤准备剪刀, 一把手术刀, 手术镊子, 两对组织镊子, 一针支架, 和15厘米 monofile 缝合不可吸收。<ol>
<li>使用热灭菌器对仪器进行消毒, 温度为240摄氏度5分钟。在使用仪器之前, 等待5分钟直到达到室温。<ol>
<li>避免任何接触的消毒仪器提示菌表面。</li>
			<li>用热器具避免皮肤灼伤。</li>
		</ol>
</li>
	</ol>
</li>
	<li>Narcotize 小鼠与异氟醚5体积% 后跟1.5 体积% 使用蒸发器和氧气流量2升/分。</li>
	<li>将鼠标放在一层纱布覆盖的升温装置上, 避免对鼠标造成热损伤。表面温度约为37摄氏度。</li>
	<li>用药膏保护眼睛。</li>
	<li>将中间背部的头发移开大约0.5 厘米2的区域, 使用剪刀或理发器, 并用适合皮肤消毒的消毒剂对无毛皮肤进行消毒。去除尽可能少的头发, 所以它保持更困难的鼠标到达伤口和线程结束后, 保持体温。</li>
	<li>用手术刀在颅尾骶部切开无毛的皮肤, 长度约为5毫米. 用钝制剂制备皮下结缔组织约1厘米深的左侧侧囊。<ol>
<li>通过踏板反射评估麻醉的水平, 并在必要的手术前调整麻醉分娩。</li>
	</ol>
</li>
	<li>用无菌组织镊子将一个10天的释放小球插入准备好的左外侧囊中。把小球放在袋子底部的平面位置。<ol>
<li>避免任何接触的颗粒到流体和湿度, 直到放置在准备袋。</li>
	</ol>
</li>
	<li>用2-3 缝线缝合皮肤。只留下非常短的螺纹末端使它更加困难为老鼠切开缝合通过啃。</li>
	<li>单独放置小鼠以减少术后窘迫, 防止伤口裂开。保持鼠标的视线, 直到它恢复了足够的意识后麻醉。 注意: 没有必要的术后疼痛管理, 因为这种干预是良好的耐受性, 没有任何迹象的不适或疼痛。另外, 我们建议外用止痛药, 如在关闭前一滴布比卡因, 这将提供多达8小时的镇痛。</li>
</ol>3. 评估模型归纳、肾病综合征和福祉的迹象
<ol><li>每天从注射日起, 通过按摩膀胱来收集晨尿 (上午08:00) 到反应杯 (1.5 毫升)。<ol>
<li>采用布拉德福德蛋白测定法测定蛋白尿, 并规范化为肌酐。 注: 对于肾病综合征的诱导, 蛋白尿应达到120毫克/毫克肌酐的阈值在7天和10后的阿霉素注射液。</li>
	</ol>
</li>
	<li>寻找腹水的发展。检查腹部周长或悬垂侧面的增加。</li>
	<li>每天早上 (上午08:00) 称鼠标、食物颗粒和饮水瓶。</li>
	<li>根据莫顿和格里菲思et, 每天至少检查一次, 使用评分表。16
</li>
</ol>4. 显色法测定尿丝氨酸蛋白酶
<ol><li>通过将15毫升无菌磷酸盐缓冲盐水 (PBS) 加入25毫克的 lyophilised 显色基质 S-2251, 制备基板工作溶液, 产生浓度为2毫米。在 PBS 中溶解抑肽酶以获得2毫克/毫升浓度的抑肽酶溶液。<ol>
<li>存储准备好的解决方案在-20 摄氏度。这大大减慢了 S-2251 的自动降解速度。不要使用带有强烈黄色阴影的旧解决方案。</li>
	</ol>
</li>
	<li>使用新鲜准备或解冻的基板工作解决方案, 并在37摄氏度热室加热。</li>
	<li>在室温下解冻尿样。<ol>
<li>避免重复解冻和冷冻, 防止蛋白酶的降解。将尿样贮存在摄氏-20 摄氏度。</li>
	</ol>
</li>
	<li>使用 96 microwell 板, 适用于光度测量。在两个井中添加3µL 的未稀释尿液, 并添加50µL 的基板工作溶液, 3 µL 的 PBS 或抑肽酶溶液的水井。用钢板封口机盖上盘子。</li>
	<li>在37摄氏度热室中孵化覆盖的 microwell 板60分钟。</li>
	<li>使用微板块读取器测量每一个井的光学密度, 在 450 nm。</li>
	<li>取无抑肽酶的 OD 与抑肽酶作为尿抑肽酶敏感的丝氨酸蛋白酶活性的差异。 注: 对具有和不带抑肽酶的配对样品的测量增加了测定的特异性, 因为基体也可以被其他不起病理生理作用的蛋白酶所切割。</li>
</ol>

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作者没有什么可透露的。

在这里, 我们证明阿霉素注射液通过球窦注射诱发实验性肾病综合征的 129S1/SvImJ 小鼠蛋白尿, 钠保留, hypoalbumenia 和高脂血症。但是, 在使用此模型时, 必须考虑到两个关键问题。首先, 模型的诱导是严格的剂量依赖性和偏差的阿霉素剂量为0.3 µg/克影响的响应小鼠15。当注射低剂量, 如14µg/克或以下, 阿霉素只导致非肾病蛋白尿, 不与显性钠保留, 腹水和高脂血症。相反, 注射更高剂?...

肾病综合征以重度蛋白尿、hypoalbuminemia、水肿、高脂血症为特征, 可视为蛋白尿肾病最极端的表现。在啮齿类动物中, 实验性肾病综合征可由单一注射 anthracyclines 或嘌呤霉素引起足细胞损伤, 类似于人的最小变化疾病和局灶节段性肾小球硬化 (FSGS) 1。在它的第一个描述以后在1955年由胡里奥·弗兰克et al。 2, 大鼠嘌呤霉素核苷肾病 (PAN) 已成为研究肾病综合征病理生理学的标准模型, 在许多研究中,3, 4, 5, 6.在小鼠中, 相应的模型可以由蒽阿霉素7诱导。然而, 有一个强大的应变依赖性, 是由至少两个遗传基因位点8决定的。此外, 肾病综合征的蛋白尿反应和病程有差异 9, 10.使用 129S1/SvImJ 小鼠和快速静脉注射阿霉素通过球窦, 蛋白尿反应达到的价值足以诱发肾病综合征的典型特点, 特别是体积保留特点腹水和几乎无钠尿7。实验性肾病综合征的钠保留被认为是在远端小管的上皮钠通道 (ENaC) 活化的结果, aberrantly 过滤丝氨酸蛋白酶, 如纤溶酶导致其γ亚单位的蛋白质降解4 ,11,12。最近, 这一概念被证明在肾病小鼠, 这是防止蛋白水解 ENaC 活化和钠保留的治疗与丝氨酸蛋白酶抑制剂抑肽酶, 同样有效的 ENaC 阻滞剂阿米洛利13。为了确保持续交付到远端小管, 抑肽酶是通过皮下植入缓释丸。未来的研究需要确定的丝氨酸蛋白酶, 负责蛋白水解 ENaC 活化的肾病综合征, 这被认为是平行的人的情况。为此, 阿霉素诱发的肾病综合征是一个有价值的模型, 可用于野生型小鼠或扩展到基因工程小鼠。其优点包括药物成本低、管理复杂性低、重现性好14。
本文通过快速静脉注射阿霉素对球窦的诱导和抑制尿丝氨酸蛋白酶的缓释微丸的植入, 证明了实验性肾病综合征用显色法测定的活动。

<table><tbody><tr><td>&lt;strong&gt;supplies&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>BD Micro FINE + U-40 (0.30 mm x 8 mm)</td><td>BD Deutschland GmbH, Heidelberg, Germany</td><td>PZN: 07468060</td><td>syring</td></tr><tr><td>ETHILON*II (5/0, 16 mm, 3/8c, 45 cm)</td><td>Johnson&amp;amp;Johnson Medical GmbH, Ethicon Deutschland, Norderstedt, Germany)</td><td>EH7823H</td><td>suture</td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;reagents&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>aprotinin (6000 KIU/mg)</td><td>LOXO, Heidelberg, Germany</td><td>CAS 9087-70-1</td><td></td></tr><tr><td>Bepanthen, Augen- und Nasensalbe</td><td>Bayer Vital GmbH, Leverkusen, Germany</td><td>PZN: 1578681</td><td>ointment</td></tr><tr><td>Chromogenix S-2251</td><td>HAEMOCHROM DIAGNOSTICA GmbH, Essen, Germany</td><td>82 0332 39</td><td>chromogenic substrate</td></tr><tr><td>doxorubicin (2.0 &amp;micro;g/&amp;micro;L)</td><td>Cell Pharm, Bad Vilbel, Germany</td><td>CAS 25316-40-9</td><td>doxorubicin</td></tr><tr><td>isofluran CP (1 mL/mL)</td><td>CP-Pharma, Burgdorf, Germany</td><td>CAS 26675-46-7</td><td>isofluran</td></tr><tr><td>pellets with matrix-driven sustained release (custom-made)</td><td>Innovative Research of America, Sarasota, FL</td><td>X-999</td><td>pellet</td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Catalog Number&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;equipment&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>ELx808 Absorbance Mikroplate Reader</td><td>BioTek, Bad Friedrichshall, Germany</td><td>ELx808</td><td>microplate reader</td></tr><tr><td>MICROSTERIL - 436</td><td>B.M.S., Trescore Balneario, Italy</td><td>GAL/436</td><td>sterilizer</td></tr><tr><td>Hybridization oven/shaker</td><td>GE Healthcare UK Limited, Amersham LIFE SCIENCE, Little Chalfont, UK</td><td>RPN 2511</td><td>heat chamber</td></tr><tr><td>Thermo MAT Pro 10 W, 15x25 cm, Lucky Reptile</td><td>Import Export Peter Hoch GmbH, Waldkirch, Germany</td><td>61201</td><td>mouse warming device</td></tr></tbody></table>

induction of nephrotic syndrome in mice by retrobulbar injection of doxorubicin and prevention of volume retention by sustained release aprotinin

肾病综合征是蛋白尿肾病最极端的表现, 其特点是重蛋白尿, hypoalbuminemia, 水肿, 由于钠保留和高脂血症。为了研究这一综合症的病理生理学, 在注射了阿霉素等有毒物质造成足细胞损伤的基础上开发了啮齿动物模型。在小鼠中, 只有很少的菌株容易受到这种模型的影响。在 wildtype 129S1/SvImJ 小鼠中, 阿霉素通过快速静脉注射到球窦, 可诱发实验性肾病综合征, 包括钠保留和水肿在内的所有人类疾病症状。蛋白尿后, 小鼠的尿丝氨酸蛋白酶活性增加, 导致上皮钠通道 (ENaC) 的活化和钠的保留。缓释抑肽酶对尿丝氨酸蛋白酶的药理抑制作用废除 ENaC 活化, 防止钠的滞留。该模型是研究 proteasuria 的病理生理学的理想方法, i. e., 活性丝氨酸蛋白酶的排泄, 导致 ENaC 活化由其γ亚基的水解。这可以看作是蛋白尿肾脏疾病中 ENaC 活化和钠保留的主要机制。

在异氟醚麻醉诱导后, 阿霉素通过左球窦快速注射。整个体积的7.25 µL/克体重注射没有任何阻力和渗出证明正确的静脉插管位置。老鼠从麻醉迅速地恢复了, 并且没有损伤在那天和尔后。特别是, 左眼没有损伤的迹象。在阿霉素注射液后, 由于阿霉素的一般毒性作用和此后恢复 (图 1a), 食物和液体摄入量在头3天下降。在注射后的头3天内, 由于不振 (<stron...

Watch this Scientific Journal Video about Induction of Nephrotic Syndrome in Mice by Retrobulbar Injection of Doxorubicin and Prevention of Volume Retention by Sustained Release Aprotinin at JoVE.com

Induction of Nephrotic Syndrome in Mice by Retrobulbar Injection of Doxorubicin and Prevention of Volume Retention by Sustained Release Aprotinin

本文介绍了阿霉素快速球注射 129S1/SvImJ 小鼠实验性肾病综合征的诱导作用。我们还治疗肾病小鼠的缓释丸含有抑肽酶, 以抑制尿丝氨酸蛋白酶活性和防止钠保留。

球注射阿霉素对小鼠肾病综合征的诱导作用及缓释抑肽酶预防容积保留率的研究

Nephrotic syndrome is the most extreme manifestation of proteinuric kidney disease and characterized by heavy proteinuria, hypoalbuminemia, and edema due ...

induction-nephrotic-syndrome-mice-retrobulbar-injection-doxorubicin

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

8.3K 次观看.  University Hospital Tübingen. 本文介绍了阿霉素快速球注射 129S1/SvImJ 小鼠实验性肾病综合征的诱导作用。我们还治疗肾病小鼠的缓释丸含有抑肽酶, 以抑制尿丝氨酸蛋白酶活性和防止钠保留。

视频: Induction of Nephrotic Syndrome in Mice by Retrobulbar Injection of Doxorubicin and Prevention of Volume Retention by Sustained Release Aprotinin

使用球后注射技术对小鼠进行有效的全身递送

Systemic Treatment for Postnatal, Juvenile, and Runted Adult Mice by Retrobulbar Sinus Injection

While tail vein injections are frequently used as a systemic route of delivery in adult mice, retrobulbar injections are an alternative method for systemic delivery with fewer limitations. First, tail vein injections (TVIs) are limited to adult mice where the size of the tail vein is suitable for access. Being restricted to treating adult mice can be problematic when dealing with mouse models that do not survive to adulthood. Second, TVIs are not feasible for mouse models with growth retardation phenotypes where the mice never achieve the size of adult wildtype mice. Therefore, retrobulbar injections can be successfully used to treat both young and small adult mice. Lastly, retrobulbar injections are performed under anesthesia, which is less stressful on the mice than TVIs that are commonly performed without anesthesia. This article presents a protocol and detailed instructions for retrobulbar injections that can be used for systemic delivery to small and young mice.

作者聚焦:通过在小鼠模型中进行球后注射克服有机酸血症基因治疗的挑战

This article provides a protocol and an accompanying video for the retrobulbar sinus injection of up to a total volume of 150 &#181;L for postnatal, juvenile, and runted adult mice. This procedure is particularly well suited for the injection of small mice (15 g) when tail vein injection is not feasible.

球后窦注射对出生后、幼年和发育成年小鼠的全身治疗

While tail vein injections are frequently used as a systemic route of delivery in adult mice, retrobulbar injections are an alternative method for ...

科研

遗传学

Subcutaneous Angiotensin II Infusion using Osmotic Pumps Induces Aortic Aneurysms in Mice

Osmotic pumps continuously deliver compounds at a constant rate into small animals. This article introduces a standard protocol used to induce aortic aneurysms via subcutaneous infusion of angiotensin II (AngII) from implanted osmotic pumps. This protocol includes calculation of AngII amount and dissolution, osmotic pump filling, implantation of osmotic pumps subcutaneously, observation after pump implantation, and harvest of aortas to visualize aortic aneurysms in mice. Subcutaneous infusion of AngII through osmotic pumps following this protocol is a reliable and reproducible technique to induce both abdominal and thoracic aortic aneurysms in mice. Infusion durations range from a few days to several months based on the purpose of the study. AngII 1,000 ng/kg/min is sufficient to provide maximal effects on abdominal aortic aneurysmal formation in male hypercholesterolemic mouse models such as apolipoprotein E deficient or low-density lipoprotein receptor deficient mice. Incidence of abdominal aortic aneurysms induced by AngII infusion via osmotic pumps is 5 - 10 times lower in female hypercholesterolemic mice and also lower in both genders of normocholesterolemic mice. In contrast, AngII-induced thoracic aortic aneurysms in mice are not hypercholesterolemia or gender-dependent. Importantly, multiple features of this mouse model recapitulate those of human aortic aneurysms.

Subcutaneous implantation of osmotic pumps provides a convenient approach for prolonged and consistent delivery of compounds. This approach has been used extensively to study both abdominal and thoracic aortic aneurysms in mice.

使用渗透皮下血管紧张素Ⅱ输液泵诱导主动脉瘤小鼠

Osmotic pumps continuously deliver compounds at a constant rate into small animals.  This article introduces a standard protocol used to induce aortic ...

医学

Assessment of Kidney Function in Mouse Models of Glomerular Disease

The use of murine models to mimic human kidney disease is becoming increasingly common. Our research is focused on the assessment of glomerular function in diabetic nephropathy and podocyte-specific VEGF-A knock-out mice; therefore, this protocol describes the full kidney work-up used in our lab to assess these mouse models of glomerular disease, enabling a vast amount of information regarding kidney and glomerular function to be obtained from a single mouse. In comparison to alternative methods presented in the literature to assess glomerular function, the use of the method outlined in this paper enables the glomerular phenotype to be fully evaluated from multiple aspects. By using this method, the researcher can determine the kidney phenotype of the model and assess the mechanism as to why the phenotype develops. This vital information on the mechanism of disease is required when examining potential therapeutic avenues in these models. The methods allow for detailed functional assessment of the glomerular filtration barrier through measurement of the urinary albumin creatinine ratio and individual glomerular water permeability, as well as both structural and ultra-structural examination using the Periodic Acid Schiff stain and electron microscopy. Furthermore, analysis of the genes dysregulated at the mRNA and protein level enables mechanistic analysis of glomerular function. This protocol outlines the generic but adaptable methods that can be applied to all mouse models of glomerular disease.

This protocol describes a full kidney work-up that should be carried out in mouse models of glomerular disease. The methods allow for detailed functional, structural, and mechanistic analysis of glomerular function, which can be applied to all mouse models of glomerular disease.

小鼠肾小球疾病模型肾脏功能的评价

The use of murine models to mimic human kidney disease is becoming increasingly common. Our research is focused on the assessment of glomerular function ...

Delivery of Exogenous Artificially Synthesized miRNA Mimic to the Kidney Using Polyethylenimine Nanoparticles in Several Kidney Disease Mouse Models

microRNAs&#160;(miRNAs), small noncoding RNAs (21-25 bases) that are not translated into proteins, inhibit lots of target messenger RNAs (mRNAs) by destabilizing and inhibiting their translation in various kidney diseases. Therefore, alternation of miRNA expression by exogenous artificially synthesized miRNA mimics is a potentially useful treatment option for inhibiting the development of many kidney diseases. However, because serum RNAase immediately degrades systematically administered exogenous miRNA mimics in vivo, delivery of miRNA to the kidney remains a challenge. Therefore, vectors that can protect exogenous miRNA mimics from degradation by RNAase and significantly deliver them to the kidney are necessary. Many studies have used viral vectors to deliver exogenous miRNA mimics or inhibitors to the kidney. However, viral vectors may cause an interferon response and/or genetic instability. Therefore, the development of viral vectors is also a hurdle for the clinical use of exogenous miRNA mimics or inhibitors. To overcome these concerns regarding viral vectors, we developed a nonviral vector method to deliver miRNA mimics to the kidney using tail vein injection of polyethylenimine nanoparticles (PEI-NPs), which led to significant overexpression of target miRNAs in several mouse models of kidney disease.

Here, we deliver exogenous artificially synthesized miRNA mimics to the kidney via tail vein injection of a nonviral vector and polyethylenimine nanoparticles in several kidney disease mouse models. This led to significant overexpression of target miRNA in the kidney, resulting in inhibited progression of kidney disease in several mouse models.

在几种肾脏疾病小鼠模型中使用聚乙烯亚胺纳米颗粒将外源人工合成的miRNA模拟物递送到肾脏

microRNAs&#160;(miRNAs), small noncoding RNAs (21-25 bases) that are not translated into proteins, inhibit lots of target messenger RNAs (mRNAs) by ...

Improved Renal Denervation Mitigated Hypertension Induced by Angiotensin II Infusion

The benefits of renal sympathetic denervation (RDN) on blood pressure have been proved in a large number of clinical trials in recent years. However, the regulatory mechanism of RDN on hypertension remains elusive. Thus, it&#39;s essential to establish a simpler RDN model in mice. In this study, osmotic mini pumps filled with Angiotensin II were implanted in 14-week-old C57BL/6 mice. One week after the implantation of the mini-osmotic pump, a modified RDN procedure was performed on bilateral renal arteries of the mice using phenol. Age-sex-matched mice were given saline and served as sham group. Blood pressure was measured at baseline and every week subsequently for 21 days. Then, renal artery, abdominal aorta and heart were collected for histological examination using H&#38;E and Masson staining. In this study, we present a simple, practical, repeatable, and standardized RDN model, which can control hypertension and alleviate cardiac hypertrophy. The technique can denervate peripheral renal sympathetic nerves without renal artery damage. Compared to previous models, the modified RDN facilitates the study of the pathobiology and pathophysiology of hypertension.

Here, we present a protocol for renal sympathetic denervation (RDN) in mice with hypertension induced by Angiotensin II infusion. The procedure is repeatable, convenient and allows to study the regulatory mechanisms of RDN on hypertension and cardiac hypertrophy.

改善肾去神经支配缓解血管紧张素II.诱导的高血压

The benefits of renal sympathetic denervation (RDN) on blood pressure have been proved in a large number of clinical trials in recent years. However, the ...

A Murine Model of Hemodialysis Access-Related Hand Dysfunction

Chronic kidney disease is a major public health problem, and the prevalence of end-stage renal disease (ESRD) requiring chronic renal replacement therapies such as hemodialysis continues to increase. Autogenous arteriovenous fistula (AVF) placement remains a primary vascular access option for ESRD patients. Unfortunately, approximately half of the hemodialysis patients experience dialysis access-related hand dysfunction (ARHD), ranging from subtle paresthesia to digital gangrene. Notably, the underlying biologic drivers responsible for ARHD are poorly understood, and no adequate animal model exists to elucidate the mechanisms and/or develop novel therapeutics for the prevention/treatment of ARHD. Herein, we describe a new mouse model in which an AVF is created between the left common iliac artery and vein, thereby facilitating the assessment of limb pathophysiology. The microsurgery includes vessel isolation, longitudinal venotomy, creation of arteriovenous anastomosis, and venous reconstruction. Sham surgeries include all the critical steps except for AVF creation. Iliac AVF placement results in clinically relevant alterations in central hemodynamics, peripheral ischemia, and impairments in hindlimb neuromotor performance. This novel preclinical AVF model provides a useful platform that recapitulates common neuromotor perturbations reported by hemodialysis patients, allowing researchers to investigate the mechanisms of ARHD pathophysiology and test potential therapeutics.

This protocol details the surgical steps of murine common iliac arteriovenous fistula creation. We developed this model to study hemodialysis access-related limb pathophysiology.

血液透析通路相关手功能障碍的小鼠模型

Chronic kidney disease is a major public health problem, and the prevalence of end-stage renal disease (ESRD) requiring chronic renal replacement ...

Drug Treatment by Central Venous Catheter in a Mouse Model of Angiotensin II Induced Abdominal Aortic Aneurysm and Monitoring by 3D Ultrasound

Since pharmaceutical treatment options are lacking in the clinical management of abdominal aortic aneurysm (AAA), animal models, in particular mouse models, are applied to advance the understanding of the disease pathogenesis and to identify potential therapeutic targets. Testing novel drug candidates to block AAA growth in these models generally requires repeated drug administration during the time course of the experiment. Here, we describe a compiled protocol for AAA induction, insertion of an intravenous catheter to facilitate prolonged therapy, and serial AAA monitoring by 3D ultrasound. Aneurysms are induced in apolipoprotein E (ApoE) deficient mice by angiotensin II release over 28 days from osmotic mini-pumps implanted subcutaneously into the mouse back. Subsequently, the surgical procedure for external jugular vein catheterization is conducted to allow for daily intravenous drug treatment or repeated blood sampling via a subcutaneous vascular access button. Despite the two dorsal implants, the monitoring of AAA development is readily facilitated by sequential semi-automated 3D ultrasound analysis, which yields comprehensive information on the expansion of aortic diameter and volume and on aneurysm morphology, as illustrated by experimental examples.

This protocol describes the consecutive implantation of an osmotic pump to induce abdominal aortic aneurysm by angiotensin II release in apolipoprotein E (ApoE) deficient mice and of a vascular access port with a jugular vein catheter for repeated drug treatment. Monitoring of aneurysm development by 3D ultrasound is effectively conducted despite dorsal implants.

血管紧张素II诱导的腹主动脉瘤小鼠模型中中心静脉导管的药物治疗及3D超声监测

Since pharmaceutical treatment options are lacking in the clinical management of abdominal aortic aneurysm (AAA), animal models, in particular mouse ...

A Mouse 5/6th Nephrectomy Model That Induces Experimental Uremic Cardiomyopathy

Chronic kidney disease (CKD) is a great risk factor for cardiovascular disease events and mortality, and progressively develops to the clinical phenotype called "uremic cardiomyopathy". We describe here an experimental CKD mouse model, named 5/6th partial nephrectomy (PNx) with pole ligation, which developed uremic cardiomyopathy at four weeks post-surgery. This PNx model was performed by a two-step surgery. In step-one surgery, both poles of the left kidney were ligated. In step-two surgery, which was performed 7 days after the step-one surgery, the right kidney was removed. For the sham surgery, the same surgery procedures were performed but without pole ligation of the left kidney or removal of the right kidney. The surgical procedures are easier and less time-consuming, compared to other methods. However, the remnant functional renal mass is not as easily controlled as the renal artery ligation. Four weeks after surgery, in comparison with the sham-operated mice, the PNx mice developed impaired renal function, anemia, cardiac hypertrophy, cardiac fibrosis, and decreased heart systolic and diastolic function.

A Mouse 5/6th Nephrectomy Model That Induces Experimental Uremic Cardiomyopathy

This manuscript provides a detailed two-step surgical procedure to perform mouse 5/6th partial nephrectomy (PNx) with pole ligation. Four weeks after surgery, in comparison with sham-operated mice, the PNx mice developed impaired renal function, anemia, cardiac hypertrophy, cardiac fibrosis, and decreased heart systolic and diastolic function.

一种诱导实验性尿毒症心肌病的小鼠 5/6th肾切除模型

Chronic kidney disease (CKD) is a great risk factor for cardiovascular disease events and mortality, and progressively develops to the clinical phenotype ...

球注射阿霉素对小鼠肾病综合征的诱导作用及缓释抑肽酶预防容积保留率的研究

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球后窦注射对出生后、幼年和发育成年小鼠的全身治疗

球后窦注射对出生后、幼年和发育成年小鼠的全身治疗

球后窦注射对出生后、幼年和发育成年小鼠的全身治疗

使用渗透皮下血管紧张素Ⅱ输液泵诱导主动脉瘤小鼠

小鼠肾小球疾病模型肾脏功能的评价

在几种肾脏疾病小鼠模型中使用聚乙烯亚胺纳米颗粒将外源人工合成的miRNA模拟物递送到肾脏

改善肾去神经支配缓解血管紧张素II.诱导的高血压

血液透析通路相关手功能障碍的小鼠模型

血管紧张素II诱导的腹主动脉瘤小鼠模型中中心静脉导管的药物治疗及3D超声监测

一种诱导实验性尿毒症心肌病的小鼠 5/6^th肾切除模型