Name: a mouse 5/6th nephrectomy model that induces experimental uremic cardiomyopathy
Uploaded: 2017-11-07T12:30:00.000+00:00
Duration: 7 min 52 s
Description: Watch this Scientific Journal Video about A Mouse 5/6th Nephrectomy Model That Induces Experimental Uremic Cardiomyopathy at JoVE.com

这项工作得到了 nih R15 1R15DK106666-01A1 (对 j. 刘) 和 nih RO1 HL071556 (J.I. 夏皮罗) 的支持。

所有动物护理和实验均由马歇尔大学机构动物护理和使用委员会 (IACUC) 根据国家卫生研究院 (NIH) 关于实验室动物的护理和使用指南获得批准。雄性 C57BL/6 小鼠 (10-12 周大) 被安置在一个无病原体的动物设施中, 在指定的房间里装有可在12小时的光照/暗循环下提供纯净空气的网箱。食物和水被提供了 ad 随意 . 
 1. 手术准备 注意: 外科手术器械和材料是从不同的来源获得的, 不特定于手术操作。其他来源的仪器和材料也可用于同一目的。有关手术器械的列表, 请参阅 材料目录 . 
<ol> <li> 手术准备 <ol> <li> 在操作表的范围内放置以下内容: 暖垫、卤素灯、氧气罐和异氟醚罐、异氟醚蒸发器、理发剪、脱毛膏、鳞片、灭菌操作套装, 消毒的手术亚麻布, 棉签, 70% 酒精, polyhydroxydine 溶液 (含1% 碘), 0.9% 氯化钠溶液, 1 毫升注射器和针头 (30 克), 3-0 和4-0 丝缝线, 丁丙诺啡和青霉素, 抗生素软膏,润滑油眼膏. </li>
		<li> 清洁一个笼子, 并在床下放置一个热垫, 供小鼠在手术后居住. </li>
	</ol> </li> </ol> 2。一步手术: 左肾极结扎术 注意: 在手术期间保持无菌状态。用无菌垫覆盖蒸压的外科器械。在手术中, 多套外科手术器械, 进行多项手术, 以防止交叉污染。如果需要再次使用仪器, 在多项手术的情况下, 用控告溶液和70% 乙醇对仪器进行消毒, 然后在器官玻璃微珠消毒器中消毒5分钟, 用70% 乙醇消毒操作区域。穿上长袍、面罩 (盖住鼻子和嘴巴)、帽 (盖头) 和一副无菌手套。每次手术后更换手套. 
<ol> <li> 清洗和消毒双手, 并佩戴无菌手术手套。戴头帽和面具. </li>
	<li> 用70% 酒精消毒桌子。将润滑剂眼膏涂抹在老鼠的双眼上, 防止麻醉时眼部干燥. </li>
	<li> 将鼠标放置在异氟醚蒸发器系统的感应室中, 并以2.5% 异氟醚100% 氧 (0.8-1.2 升/分) 的混合物诱导麻醉, 用于 1-2 min. 
	注: 麻醉的功效是由捏尾引起的疼痛反射, 以及眼睑没有眼睑反射而决定的. </li>
	<li> 注射青霉素 (4万单位/千克体重, IM) 和丁丙诺啡 (0.02 毫克/千克体重, SC), 以防止感染和疼痛的手术过程中. </li>
	<li> 将鼠标放在暖垫 (最初设置在42和 #176; c) 上, 用直肠温度计来保持和监测37和 #176 的体温; 在手术过程中。在手术过程中, 将小鼠的鼻子放入鼻锥, 用 0.8-1.2% 的异氟醚与100% 氧 (0.8-1.2 升/分) 的混合物进行通气. </li>
	<li> 将鼠标放在右侧。用理发剪刮掉手术区的毛发, 然后用脱毛膏除去。用 polyhydroxydine 溶液 (含1% 碘) 消毒现场, 然后用70% 乙醇。将蒸压的外科亚麻布对准手术区. </li>
	<li> 做一个〜1.0 厘米腰椎切口和分离的肌肉和筋膜暴露左肾使用手术刀和钳 ( 图 1a ). </li>
	<li> 通过用小钝钳拉动肾脂肪轻轻拉出左肾。在这个过程中, 为了不扰乱肾上腺, 用镊子将肾上腺从肾脏向上移. </li>
	<li> 使用钝钳, 轻轻地从肾脏的上端将结缔组织和肾上腺从肾的上部向肾的中间的幽门处, 血管进入和出口。识别和隔离输尿管. 
	注: 输尿管位于肾脏的下半部, 在结缔组织内, 在光下清晰可见. </li>
	<li> 轻轻地抓住结缔组织并从肾脏组织中分离, 以确保输尿管没有结扎. </li>
	<li> 在极性结扎之前, 使用无菌尺来鉴别肾脏的1/3 部分的优劣。老鼠肾脏通常是大约 1.25-1.5 cm 长. </li>
	<li> 结扎使用3-0 丝线缝合线和适当的力量, 大约0.4 厘米从上级部分往幽门的中部和相似地, 0.4 cm 从下等部分对幽门 ( 图 1a , 点式行). </li>
	<li> 在结扎两个极点 ( 图 1b ) 后, 允许2分钟观察时间以确保没有出血. 
	注意: 在这个过程中, 避免结扎肾上腺、肾动脉和输尿管是非常重要的。如果左输尿管被结扎, 老鼠将在第二步手术中切除右肾后死亡。结扎力可以限制血流到两极, 而不会引起肾出血。在结扎后的 1-2 分钟内, 两极因血液流向两极的极限而变色. </li>
	<li> 将左肾推回到原来的位置。分别用4-0 和3-0 丝缝合线将肌肉和皮肤闭合。将抗生素软膏应用于手术区, 以更好的恢复和消毒. </li>
	<li> 结扎后, 将鼠标放在笼中, 将其放在一个带有热灯的笼子中, 使寝具区域接近37和 #176; C (用温度计监测), 然后将动物放在单独的笼子里. 
	注意: 监测鼠标, 直到它已经恢复足够的意识, 以维护胸骨卧床. </li>
	<li> 允许在恢复过程中方便地获得水和食物。管理青霉素和丁丙诺啡, 每12小时后头3天, 以防止感染和疼痛. 
	注意: 老鼠应该在 2-3 天内从手术创伤中恢复过来, 并且应该回到正常的饮食、饮酒和运动中。此时, 将鼠标逐个进行完全恢复。必须对以下情况进行监测: 饮酒、进食、行走方式、笨拙的步态、驼背、眼部和/或鼻腔分泌物以及攻击性行为。安乐如果它显示任何癫痫发作、昏迷、无法治愈的感染、行走困难、步态丧失、无法进食和饮水, 以及15% 的术后体重损失. </li>
</ol> 3。第二步手术: 右肾切除术 <ol> <li> 七天后, 在步骤2.1 到2.10 中将右肾暴露为左肾, 除非将鼠标放在其左侧. </li>
	<li> 将右肾放在亚麻布上, 用钝钳将周围的脂肪和结缔组织清除干净. </li>
	<li> 识别肾动脉和静脉, 然后将两个连字 (3-0 丝) 分别放置在血管周围的一个松散结。轻轻地挑逗结缔组织和肾上腺腺体从肾脏的上端向中间的幽门的肾脏使用钝钳。轻轻地将输尿管从下端移出, 并将所有的血管系在一起 (3-0 丝) 而不会造成出血. </li>
	<li> 将两个松散的连字节沿容器移动。一个朝向腹主动脉一侧, 另一个朝向肾侧. </li>
	<li> 首先将结扎结 (朝向腹主动脉侧) 与双节结; 实心结会改变右肾的颜色。将结扎结 (朝向肾脏一侧) 系成双结。在这两个结之间留出足够的空间, 以便在两节之间切断血管而不割断结扎结. </li>
	<li> 切开两节之间的肾血管, 取出右肾。检查肾血管是否可能出血. </li>
	<li> 用无菌纱布烘干右肾, 称其为重. </li>
	<li> 用4-0 丝缝合线关闭肌肉切口, 然后用3-0 丝缝合线闭合皮肤切口. </li>
	<li> 将鼠标分别放在一个干净的单独的笼子里, 在笼子上放一盏热灯 (步骤 2.15), 以方便地获得水和食物, 至少24小时才能恢复。管理青霉素和丁丙诺啡每12小时3天, 以防止感染和疼痛. 
	注意: 老鼠应该在 2-3 天内从手术创伤中恢复过来, 并且应该回到正常的饮食、饮酒和运动中. </li>
	<li> 手术后, 每天两次监测小鼠三天, 然后每天一次进行实验。遵循步骤2.16 中描述的后监视和处理程序. </li>
</ol> 4。假手术 <ol> <li> 为假手术, 执行相同的手术程序, 包括肾脏暴露, 组织解剖, 伤口闭合, 但没有左肾极结扎或右肾切除. </li> </ol>
 5. 实验性尿毒症心肌病的评估 <ol> <li> 在牺牲前一天, 执行经胸超声心动图和捕获图像 18 。
	注: 在这里, 经胸超声心动图进行了鼠标处理表和直肠温度计。超声心动图图像被捕获使用18-38 兆赫工作频率传感器连接到一个成像系统。根据我们描述的 18 , 计算出相对壁厚 (RWT)、心肌性能指数 (MPI) 和左心室质量指数 (LVMI). </li>
	<li> 在牺牲时, 测量体重。根据制造商和 #39 的指示, 用一台小贩的离心机测量红细胞压积 (小贩)。用肝素包膜管制备血浆样品, 离心10分钟, 在 1500 x g 在4和 #176; C 在台式冷藏离心机。根据制造商和 #39 的说明, 使用套件测量胱 C、肌酐和发髻。重复执行测量. </li>
	<li> 在实验结束时, 麻醉用氯胺酮 (90 毫克/千克, ip) 和 xylaxine (10 毫克/千克, ip) 和切口的牺牲。从胸部取出后测量心脏重量。准备心脏 (LV) 和肾脏匀, 以及 Type-1 胶原蛋白 (COL-1) 的测定, 磷酸化的 c Src 和蛋白质羰基, 如前所述, 18 , 19 . </li>
</ol>

<ol>
	<li>Sarnak, M. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kidney+Disease+as+a+Risk+Factor+for+Development+of+Cardiovascular+Disease:+A+Statement+From+the+American+Heart+Association+Councils+on+Kidney+in+Cardiovascular+Disease,+High+Blood+Pressure+Research,+Clinical+Cardiology,+and+Epidemiology+and+Prevention.">Kidney Disease as a Risk Factor for Development of Cardiovascular Disease: A Statement From the American Heart Association Councils on Kidney in Cardiovascular Disease, High Blood Pressure Research, Clinical Cardiology, and Epidemiology and Prevention.</a> Circulation. 108 (17), 2154-2169 (2003).</li><li>Levey, A. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlling+the+epidemic+of+cardiovascular+disease+in+chronic+renal+disease:+what+do+we+know?+What+do+we+need+to+learn?+Where+do+we+go+from+here?+National+Kidney+Foundation+Task+Force+on+Cardiovascular+Disease.">Controlling the epidemic of cardiovascular disease in chronic renal disease: what do we know? What do we need to learn? Where do we go from here? National Kidney Foundation Task Force on Cardiovascular Disease.</a> Am J Kidney Dis. 32 (5), 853-906 (1998).</li><li>Schiffrin, E. L., Lipman, M. L., Mann, J. F. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+Kidney+Disease.">Chronic Kidney Disease.</a> Effects on the Cardiovascular System. 116 (1), 85-97 (2007).</li><li>Foley, R. N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+and+echocardiographic+disease+in+patients+starting+end-stage+renal+disease+therapy.">Clinical and echocardiographic disease in patients starting end-stage renal disease therapy.</a> Kidney Int. 47 (1), 186-192 (1995).</li><li>Foley, R. N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+prognostic+importance+of+left+ventricular+geometry+in+uremic+cardiomyopathy.">The prognostic importance of left ventricular geometry in uremic cardiomyopathy.</a> J Am Soc Nephrol. 5 (12), 2024-2031 (1995).</li><li>Alhaj, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Uremic+cardiomyopathy:+an+underdiagnosed+disease.">Uremic cardiomyopathy: an underdiagnosed disease.</a> Congest Heart Fail. 19 (4), 40-45 (2013).</li><li>Gulati, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Association+of+fibrosis+with+mortality+and+sudden+cardiac+death+in+patients+with+nonischemic+dilated+cardiomyopathy.">Association of fibrosis with mortality and sudden cardiac death in patients with nonischemic dilated cardiomyopathy.</a> JAMA. 309 (9), 896-908 (2013).</li><li>London, G. M., Parfrey, P. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiac+disease+in+chronic+uremia:+pathogenesis.">Cardiac disease in chronic uremia: pathogenesis.</a> Adv Ren Replace Ther. 4 (3), 194-211 (1997).</li><li>Mohmand, B., Malhotra, D. K., Shapiro, J. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Uremic+cardiomyopathy:+role+of+circulating+digitalis+like+substances.">Uremic cardiomyopathy: role of circulating digitalis like substances.</a> Front Biosci. 10, 2036-2044 (2005).</li><li>Himmelfarb, J., McMonagle, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Manifestations+of+oxidant+stress+in+uremia.">Manifestations of oxidant stress in uremia.</a> Blood Purif. 19 (2), 200-205 (2001).</li><li>Okamura, D. M., Himmelfarb, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tipping+the+redox+balance+of+oxidative+stress+in+fibrogenic+pathways+in+chronic+kidney+disease.">Tipping the redox balance of oxidative stress in fibrogenic pathways in chronic kidney disease.</a> Pediatr Nephrol. 24 (12), 2309-2319 (2009).</li><li>Himmelfarb, J., Stenvinkel, P., Ikizler, T. A., Hakim, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+elephant+in+uremia:+oxidant+stress+as+a+unifying+concept+of+cardiovascular+disease+in+uremia.">The elephant in uremia: oxidant stress as a unifying concept of cardiovascular disease in uremia.</a> Kidney Int. 62 (5), 1524-1538 (2002).</li><li>Becker, B. N., Himmelfarb, J., Henrich, W. L., Hakim, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reassessing+the+cardiac+risk+profile+in+chronic+hemodialysis+patients:+a+hypothesis+on+the+role+of+oxidant+stress+and+other+non-traditional+cardiac+risk+factors.">Reassessing the cardiac risk profile in chronic hemodialysis patients: a hypothesis on the role of oxidant stress and other non-traditional cardiac risk factors.</a> J Am Soc Nephrol. 8 (3), 475-486 (1997).</li><li>Drummond, C. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reduction+of+Na/K-ATPase+affects+cardiac+remodeling+and+increases+c-kit+cell+abundance+in+partial+nephrectomized+mice.">Reduction of Na/K-ATPase affects cardiac remodeling and increases c-kit cell abundance in partial nephrectomized mice.</a> Am J Physiol Heart Circ Physiol. 306 (12), 1631-1643 (2014).</li><li>Kennedy, D. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Partial+nephrectomy+as+a+model+for+uremic+cardiomyopathy+in+the+mouse.">Partial nephrectomy as a model for uremic cardiomyopathy in the mouse.</a> Am J Physiol Renal Physiol. 294 (2), 450-454 (2008).</li><li>Haller, S. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monoclonal+antibody+against+marinobufagenin+reverses+cardiac+fibrosis+in+rats+with+chronic+renal+failure.">Monoclonal antibody against marinobufagenin reverses cardiac fibrosis in rats with chronic renal failure.</a> Am J Hypertens. 25 (6), 690-696 (2012).</li><li>Tian, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spironolactone+attenuates+experimental+uremic+cardiomyopathy+by+antagonizing+marinobufagenin.">Spironolactone attenuates experimental uremic cardiomyopathy by antagonizing marinobufagenin.</a> Hypertension. 54 (6), 1313-1320 (2009).</li><li>Liu, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Attenuation+of+Na/K-ATPase+Mediated+Oxidant+Amplification+with+pNaKtide+Ameliorates+Experimental+Uremic+Cardiomyopathy.">Attenuation of Na/K-ATPase Mediated Oxidant Amplification with pNaKtide Ameliorates Experimental Uremic Cardiomyopathy.</a> Sci Rep. 6, 34592 (2016).</li><li>Yan, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Involvement+of+reactive+oxygen+species+in+a+feed-forward+mechanism+of+Na/K-ATPase-mediated+signaling+transduction.">Involvement of reactive oxygen species in a feed-forward mechanism of Na/K-ATPase-mediated signaling transduction.</a> J Biol Chem. 288 (47), 34249-34258 (2013).</li><li>Leelahavanichkul, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Angiotensin+II+overcomes+strain-dependent+resistance+of+rapid+CKD+progression+in+a+new+remnant+kidney+mouse+model.">Angiotensin II overcomes strain-dependent resistance of rapid CKD progression in a new remnant kidney mouse model.</a> Kidney Int. 78 (11), 1136-1153 (2010).</li><li>Ma, L. J., Fogo, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Model+of+robust+induction+of+glomerulosclerosis+in+mice:+importance+of+genetic+background.">Model of robust induction of glomerulosclerosis in mice: importance of genetic background.</a> Kidney Int. 64 (1), 350-355 (2003).</li></ol>

作者没有什么可透露的。

大鼠 5/6th PNx 模型被广泛用于研究 CKD。由于小鼠的肾脏体积较小, 传统的动脉结扎和极切除术在小鼠模型中具有很大的挑战性, 可能造成高死亡率和意外出血/失血。
我们采用了小鼠 PNx 模型与极结扎, 以克服出血/失血。该 PNx 模型耗时少, 成活率高, 重现性好。这种极结扎模型在四周后时发展了人尿毒症心肌病的表型变化, 从而为研究其机制和治疗靶点提供了一个模型。
在这个 PNx 模型中, 5/6th PNx 是肾脏质量降低的一种近似估计, 至今为止, 还没有准确计算出两个不同大小/质量的肾脏残余肾脏质量的方法。但是, 结扎应以非常相似的方式进行, 以减少手术的变异。我们估计, 保持约40% 的左肾 (或约20% 的总肾脏质量) 可以显著降低死亡率 (约 20%), 而不牺牲的发展, 尿毒症型心肌病表型。另一个问题是结扎的力量需要练习。
与 5/6th PNx 模型通过极切除或肾动脉结扎1520相比, 极结扎法简便、快速, 且为要求较低。此 PNx 模型的可能优点包括: 1) 在 PNx 模型中, 通过极切除或动脉结扎, 2), 避免可能的出血, 从而使炎症反应引起的氧化应激减弱极结扎, 3) 更低的死亡率 (约 20%), 和 4) 快速发展的尿毒症心肌病表型。成功的极套结扎显示了连续的不可逆损伤和被结扎的极点的退化。在两周的后 (图 4) 中, 被结扎的极点是可见的, 并且主要在四周后 (图 5) 消失。比较两周和四周后的结扎极点, 表明对极结扎的炎症反应。
这种 PNx 模型可能有助于研究药物作用机制和药物筛选。我们最近使用这个 PNx 模型来研究钠/钾 atp 酶信号和氧化应激在尿毒症心肌病的发展中的作用。pNaKtide 是钠/钾 atp 酶信号的特异拮抗剂, 它不仅能防止尿毒症心肌病的发生, 而且还能逆转尿毒症心肌病的发生18。
该协议中的关键步骤是 follows:1) 一个更积极的结扎 (残余左肾肿块小于 30%) 将显著增加死亡率 (约 50%). 2) 结扎的力量需要实践达到的地步, 结扎限制血液供应到极点, 而不破坏胶囊和肾脏组织。打破肾脏组织导致肾出血, 可能导致死亡后, 右肾切除。一个成功的结扎杆在结扎后 1-2 分钟内变色, 由于血流到极点的限制。3) 需要特别注意避免结扎左肾的输尿管或在结扎两极时损害输尿管。在切除右肾后, 有结扎或损伤的输尿管的老鼠将无法存活。这可能是最明显的缺点的极点结扎方法。4) 在手术过程和后护理中保持体温。5) 用抗生素控制可能的感染。

CKD, 也被称为慢性肾衰竭, 是随着时间的推移肾脏功能的逐步丧失, 最终发展成永久性肾衰竭。CKD, 从早期肾脏疾病状态到终末期肾脏疾病 (末期), 是心血管疾病事件和死亡率的一个很大的危险因素, 并逐步发展到临床表型称为 "尿毒症心肌病"1, 2,3。CKD 或末期患者的尿毒症心肌病与心血管异常有关, 主要是由于左心室 (lv) 压力和/或容积超负荷导致的, 导致 lv 肥厚、lv 扩张和 lv 收缩功能障碍4 ,5,6。心脏纤维化是尿毒症心肌病的另一种常见的病理过程, 它能降低心脏顺应性, 导致左心室舒张功能障碍。严重的心脏纤维化可能导致心脏猝死甚至在那些没有心脏症状的患者中7。
5/6th PNx 是一种常用的 CKD 动物模型, 用于动物研究, 包括肾衰竭、尿毒症心肌病和高血压。PNx 是通过消融5/6 肾实质来实现的。大鼠模型最初是用两种最常用的手术切除或梗塞的方法开发的。大鼠 PNx 模型是一个非常有用的模型来研究尿毒症心肌实质升高血压, 心肌肥厚和舒张功能受损。后来, 老鼠 PNx 模型, 操作与相似的技术作为鼠模型, 被开发了由于广泛的可及性在小鼠系统做基因操纵。
有文献记载, 全身氧化剂应激是临床和实验性尿毒症心肌病的一个恒定特征8,9。此外, 氧化剂的压力有助于尿毒症综合征10, 并在尿毒症心肌病的发病机制中起关键作用111213。在这一点上, 我们已经证明了啮齿动物 5/6th PNx 模型导致尿毒症心肌病的生理、形态和生化特征1415,16, 17. 在这里描述的鼠标 PNx 模型中, PNx 操作的小鼠产生了明显的氧化应激, 至少部分由钠/钾 atp 酶信号传导, 这在 PNx 介导的尿毒症心肌病中是至关重要的。钠/钾 atp 酶信号的衰减不仅降低了氧化放大, 而且还改善了 PNx 介导的实验性尿毒症心肌病的型变化18。

<table><tbody><tr><td>Iris Scissors, 11.5 cm, Straight</td><td>World Precision Instruments</td><td>501758</td><td></td></tr><tr><td>WPI Swiss Tweezers #5 11 cm, 0.1x0.06 mm Tips</td><td>World Precision Instruments</td><td>504506</td><td></td></tr><tr><td>Jewelers #5 Forceps, 11cm, Straight, Titanium</td><td>World Precision Instruments</td><td>555227F</td><td></td></tr><tr><td>Iris Forceps, 10cm, Straight, Serrated</td><td>World Precision Instruments</td><td>15914</td><td></td></tr><tr><td>Tweezers #3, 11cm, 0.2x0.4mm Tips</td><td>World Precision Instruments</td><td>501976</td><td></td></tr><tr><td>Medesy Iris 4.5&quot; Curved Scissors, Stainless Steel</td><td>Net23</td><td>3512</td><td>www.Net32.com</td></tr><tr><td>Dr. Slick Iris Scissors; 3.5&quot;</td><td>Avid Max</td><td>220-1-965-IrisScsrs-P&amp;nbsp;</td><td>www.Avid Max.com</td></tr><tr><td>Miltex Iris Scissors 4 1/8&quot; Curved</td><td>4mdmedical</td><td>V95-306</td><td>www.4mdmedical.com</td></tr><tr><td>5&quot; Hemostat clap, curved jaw</td><td>PJTool</td><td>4355</td><td>www.pjtool.com</td></tr><tr><td>sklar Knapp Iris scissors, straight and sharp/blunt 4&quot;</td><td>Medical Device depot</td><td>64-3430</td><td>www.medicaldevicedepot.com</td></tr><tr><td>Kelly Hemostatic Forceps straight 5.5&quot;</td><td>Pilgtimmedical</td><td>FA710-50</td><td>www.pilgtimmedical.com</td></tr><tr><td>C57BL/6 mice</td><td>Hilltop Lab Animals Inc.</td><td></td><td></td></tr><tr><td>&amp;nbsp;Mouse Handling Table and rectal thermometer</td><td>Visualsonics</td><td></td><td></td></tr><tr><td>MicroScan transducer&amp;nbsp;</td><td>Vevo 1100</td><td>MS400</td><td></td></tr><tr><td>Vevo 1100 Imaging System&amp;nbsp;</td><td>FUJIFILM VisualSonics Inc.</td><td></td><td></td></tr><tr><td>Cystatin C ELISA kit and mouse creatinine kit&amp;nbsp;</td><td>Crystal Chem. Inc.</td><td></td><td></td></tr><tr><td>Mouse BUN ELISA kit&amp;nbsp;</td><td>MyBioSource Inc&amp;nbsp;</td><td></td><td></td></tr></tbody></table>

a mouse 5/6th nephrectomy model that induces experimental uremic cardiomyopathy

慢性肾脏疾病 (CKD) 是心血管疾病事件和死亡率的一个很大的危险因素, 并逐步发展到临床表型称为 "尿毒症心肌病"。我们在这里描述一个实验性 CKD 小鼠模型, 命名为 5/6th部分肾切除 (PNx) 与极结扎, 在四周后的发展为尿毒症心肌病。这种 PNx 模型是由 two-step 手术进行的。在第一步手术中, 左肾的两极都结扎了。在第一步手术7天后进行的两次手术中, 右肾切除。对于假手术, 同样的手术程序进行, 但没有极结扎左肾或切除右肾。与其他方法相比, 手术过程更容易、更耗时。然而, 残余功能性肾肿块不像肾动脉结扎那样容易控制。手术后四周, 与假手术小鼠相比, PNx 小鼠肾功能受损, 贫血, 心肌肥厚, 心脏纤维化, 心脏收缩和舒张功能下降。

数据表明, 此修改后的 5/6 th PNx 模型是一种简单有效的研究尿毒症心肌病的模型。在四周后, 该 PNx 模型显示肾功能受损, 贫血, 心肌肥厚, 心脏纤维化, 并降低心脏收缩和舒张功能。结果概述如下。
在四周的后, PNx 小鼠除了心脏形态学和生化变化, 与人类尿毒症心肌病的表型相一致外, 还发育受损的肾功能。这些 PNx 介导的变化包括以下观察与假操作小鼠相比。首先, 正如预期的那样, PNx 手术引起了肾功能的严重损害。如图 2所示, PNx 手术显著增加了胱 C 的血浆水平 (图 2a, n = 16), 肌酐 (图 2b, n = 16), 发髻 (图 2C, n = 16), 当与虚假手术.其次, 在 PNx 小鼠 (图 2d, n=8) 中, 有明显的低 PNx 的受激性贫血。第三, PNx 介导的心肌肥厚和舒张功能障碍表现为增加的心脏重量/体重比 (图 3a), 和经胸超声心动图 (回声) 分析显示显着增加后壁厚度 (PWT)、前壁厚度 (AWT)、RWT (图 3b)、MPI (图 3c) 和 LVMI (图 3d)。射血分数没有显著变化 (EF,%)。第四, PNx 介导的心肌纤维化通过 PNx 诱导的 i 型胶原蛋白表达在 LV 匀 (图 4a) 和左残余肾脏匀 (图 4b) 中显示。第五, 氧化剂应激在实验性尿毒症心肌病中起重要作用。在这个 PNx 模型中, PNx 在 LV 匀中显著地刺激了蛋白质的直接蛋白羰化修饰 (图 4c) 和 c Src 激活 (图 4d)。PNx 手术并没有造成四周后的体重显著下降。然而, PNx 模型采用极结扎法并没有诱发高血压。在一步手术前的一天和一天前的一项治疗中, PNx 小鼠的收缩期血压、舒张 bp 和平均 bp (未显示的数据) 之间没有显著差异, 这与 C57BL/6 鼠菌株的概念是一致的。未建立高血压模型20,21。数据还表明, 这种 PNx 模型不太可能是急性肾衰竭的模型。此外, PNx 介导的心脏改变, 但不损害肾功能, 通过阻断 c 型钢 Src 活化或诱导血红素 oxygenase-1 (HO-1)18。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/55825/55825fig1.jpg"> 
图 1: 左肾的极结扎.照片显示左肾结扎 (左) 和结扎后 (右)。左图中的被溺爱的线条显示了估计的结扎线。<a href="//ecsource.jove.com/files/ftp_upload/55825/55825fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/55825/55825fig2.jpg"> 
图 2: PNx 与极结扎导致肾功能受损和贫血.在四周后, PNx 手术显着增加血浆水平 (a) 胱 C (n = 16), (b) 肌酐 (n = 16), (C) 发髻 (n = 16), 与假手术相比。PNx 也明显减少 (d) 红细胞压积 (小贩) 在 PNx 小鼠 (n = 8)。数据被表示为平均± SEM. **, p 和 #60; 0.01, 假手术小鼠与 PNx 操作的小鼠。<a href="//ecsource.jove.com/files/ftp_upload/55825/55825fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/55825/55825fig3.jpg"> 
图 3: PNx 与极结扎刺激心肌肥大.在四周后, PNx 手术显著增加 (a) 的心脏重量/体重比 (a, n = 8), (b) 相对壁厚 (RWT, n = 1 9-21), (c) 心肌性能指数 (MPI, n = 19-21), 以及(d) 左心室质量指数 (LVMI, n = 19-21)。数据被表示为平均± SEM. **, p 和 #60; 0.01, 假手术小鼠与 PNx 操作的小鼠。<a href="//ecsource.jove.com/files/ftp_upload/55825/55825fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/55825/55825fig4.jpg"> 
图 4: PNx 与极结扎刺激 i 型胶原蛋白表达, 蛋白质羰基化和 c-Src 活化.在四周后, PNx 刺激 i 型胶原蛋白表达在左心室 (LV) 匀 (a, n=11) 和左残余肾脏匀 (b, n = 7-10)。也在 LV 匀, PNx 刺激直接蛋白质羰基化 (c, n = 6) 和 c Src 活化 (d, n = 6)。数据被表示为平均± SEM. **, p 和 #60; 0.01, 假手术小鼠与 PNx 操作的小鼠。<a href="//ecsource.jove.com/files/ftp_upload/55825/55825fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>

Watch this Scientific Journal Video about A Mouse 5/6th Nephrectomy Model That Induces Experimental Uremic Cardiomyopathy at JoVE.com

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

这份手稿提供了一个详细的 two-step 手术程序执行小鼠 5/6th部分肾切除 (PNx) 与极结扎。手术后四周, 与假手术小鼠相比, PNx 小鼠肾功能受损, 贫血, 心肌肥厚, 心脏纤维化, 心脏收缩和舒张功能下降。

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 ...

a-mouse-56th-nephrectomy-model-that-induces-experimental-uremic

Universidad San Sebastian

Research

JoVE Journal

Biology

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

20.3K 次观看.  Marshall University. 这份手稿提供了一个详细的 two-step 手术程序执行小鼠 5/6th部分肾切除 (PNx) 与极结扎。手术后四周, 与假手术小鼠相比, PNx 小鼠肾功能受损, 贫血, 心肌肥厚, 心脏纤维化, 心脏收缩和舒张功能下降。

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

Normothermic Cardiac Arrest and Cardiopulmonary Resuscitation: A Mouse Model of Ischemia-Reperfusion Injury

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
frequently caused by whole-body hypoperfusion.5,6 Successful reproduction of whole-body hypoperfusion in rodent models has been fraught with 
difficulty.7-9,9,10 Models which employ focal ischemia have repeatedly demonstrated results which do not translate to the clinical setting, and larger 
animal models which allow for whole body hypoperfusion lack access to the full toolset of genetic manipulation possible in the mouse.11,12 However, in recent 
years a mouse model of cardiac arrest and cardiopulmonary resuscitation has emerged which can be adapted to model AKI.13 This model reliably reproduces 
physiologic, functional, anatomic, and histologic outcomes seen in clinical AKI, is rapidly repeatable, and offers all of the significant advantages of a murine surgical 
model, including access to genetic manipulative techniques, low cost relative to large animals, and ease of use. Our group has developed extensive experience 
with use of this model to assess a number of organ-specific outcomes in AKI.14,15

A powerful model for perioperative and critical care related acute kidney injury is presented. Using whole body hypoperfusion induced by cardiac arrest it is possible to nearly replicate the histologic and functional changes of clinical AKI.

常温心脏骤停和心肺复苏术：缺血再灌注损伤的小鼠模型

Acute Kidney Injury (AKI) is a common, highly lethal, complication of critical illness which has a high mortality1-4 and which is most 
frequently caused ...

科研

医学

Surgical Techniques for Catheter Placement and 5/6 Nephrectomy in Murine Models of Peritoneal Dialysis

Peritoneal dialysis (PD) is a renal replacement therapy consistent on the administration and posterior recovery of a hyperosmotic fluid in the peritoneal cavity to drain water and toxic metabolites that functionally-insufficient kidneys are not able to eliminate. Unfortunately, this procedure deteriorates the peritoneum. Tissue damage triggers the onset of inflammation to heal the injury. If the injury persists and inflammation becomes chronic, it may lead to fibrosis, which is a common occurrence in many diseases. In PD, chronic inflammation and fibrosis, along with other specific processes related to these ones, lead to ultrafiltration capacity deterioration, which means the failure and subsequent cessation of the technique. Working with human samples provides information about this deterioration but presents technical and ethical limitations to obtain biopsies. Animal models are essential to study this deterioration since they overcome these shortcomings.
A chronic mouse infusion model was developed in 2008, which benefits from the wide range of genetically modified mice, opening up the possibility of studying the mechanisms involved. This model employs a customized device designed for mice, consisting of a catheter attached to an access port that is placed subcutaneously at the back of the animal. This procedure avoids continuous puncture of the peritoneum during long-term experiments, reducing infections and inflammation due to injections. Thanks to this model, peritoneal damage induced by chronic PD fluid exposure has been characterized and modulated. This technique allows the infusion of large volumes of fluids and could be used for the study of other diseases in which inoculation of drugs or other substances over extended periods of time is necessary.
This article shows the method for the surgical placement of the catheter in mice. Moreover, it explains the procedure for a 5/6 nephrectomy to mimic the state of renal insufficiency present in PD patients.

This article shows the method for the surgical placement in mice of an intraperitoneal catheter attached to an access port that is positioned at the back of the animal. Moreover, it explains the procedure for a 5/6 nephrectomy to resemble the uremic state of PD patients.

小鼠腹膜透析模型导管置入及5/6 切除术的手术技术

Peritoneal dialysis (PD) is a renal replacement therapy consistent on the administration and posterior recovery of a hyperosmotic fluid in the peritoneal ...

工程学

Quantitative Real-Time PCR Evaluation of microRNA Expressions in Mouse Kidney with Unilateral Ureteral Obstruction

MicroRNAs (miRNAs) are single stranded, non-coding RNA molecules that typically regulate gene expression at the post-transcriptional level by binding to partially complementary target sites in the 3&#39; untranslated region (UTR) of messenger RNA (mRNA), which reduces the mRNA&#39;s translation and stability. The miRNA expression profiles in various organs and tissues of mice have been investigated, but standard methods for the purification and quantification of miRNA in mouse kidney have not been available. We have established an effective and reliable method for extracting and evaluating miRNA expression in mouse kidney with renal interstitial fibrosis by quantitative reverse-transcription polymerase chain reaction (qRT-PCR). The protocol required five steps: (1) creation of sham and unilateral ureteral obstruction (UUO) mice; (2) extraction of kidney samples from the UUO mice; (3) extraction of total RNA, which includes miRNA, from the kidney samples; (4) complementary DNA (cDNA) synthesis with reverse transcription from miRNA; and (5) qRT-PCR using the cDNA. Using this protocol, we successfully confirmed that compared to the controls, the expression of miRNA-3070-3p was significantly increased and those of miRNA-7218-5p and miRNA-7219-5p were significantly decreased in the kidneys of a mouse model of renal interstitial fibrosis. This protocol can be used to determine the miRNA expression in the kidneys of mice with UUO.

We describe a method for evaluating the microRNA expression in the kidneys of mice with unilateral ureteral obstruction (UUO) by quantitative reverse-transcription polymerase chain reaction. This protocol is suitable for studying kidney microRNA expression profiles in mice with UUO and in the context of other pathological conditions.

小鼠肾中微RNA表达与单体尿阻的定量实时PCR评价

MicroRNAs (miRNAs) are single stranded, non-coding RNA molecules that typically regulate gene expression at the post-transcriptional level by binding to ...

遗传学

5/6th Nephrectomy in Combination with High Salt Diet and Nitric Oxide Synthase Inhibition to Induce Chronic Kidney Disease in the Lewis Rat

Chronic kidney disease (CKD) is a global problem. Slowing CKD progression is a major health priority. Since CKD is characterized by complex derangements of homeostasis, integrative animal models are necessary to study development and progression of CKD. To study development of CKD and novel therapeutic interventions in CKD, we use the 5/6th nephrectomy ablation model, a well known experimental model of progressive renal disease, resembling several aspects of human CKD. The gross reduction in renal mass causes progressive glomerular and tubulo-interstitial injury, loss of remnant nephrons and development of systemic and glomerular hypertension. It is also associated with progressive intrarenal capillary loss, inflammation and glomerulosclerosis. Risk factors for CKD invariably impact on endothelial function. To mimic this, we combine removal of 5/6th of renal mass with nitric oxide (NO) depletion and a high salt diet. After arrival and acclimatization, animals receive a NO synthase inhibitor (NG-nitro-L-Arginine) (L-NNA) supplemented to drinking water (20 mg/L) for a period of 4 weeks, followed by right sided uninephrectomy. One week later, a subtotal nephrectomy (SNX) is performed on the left side. After SNX, animals are allowed to recover for two days followed by LNNA in drinking water (20 mg/L) for a further period of 4 weeks. A high salt diet (6%), supplemented in ground chow (see time line Figure 1), is continued throughout the experiment. Progression of renal failure is followed over time by measuring plasma urea, systolic blood pressure and proteinuria. By six weeks after SNX, renal failure has developed. Renal function is measured using 'gold standard' inulin and para-amino hippuric acid (PAH) clearance technology. This model of CKD is characterized by a reduction in glomerular filtration rate (GFR) and effective renal plasma flow (ERPF), hypertension (systolic blood pressure&gt;150 mmHg), proteinuria (&gt; 50 mg/24 hr) and mild uremia (&gt;10 mM). Histological features include tubulo-interstitial damage reflected by inflammation, tubular atrophy and fibrosis and focal glomerulosclerosis leading to massive reduction of healthy glomeruli within the remnant population (&lt;10%). Follow-up until 12 weeks after SNX shows further progression of CKD.

A two-stage method to establish chronic kidney disease (CKD) in the Lewis rat by surgically removing 5/6th of renal mass is described. Combination of the surgical procedure, NOS-inhibition and a high-salt diet leads to a model resembling human CKD, allowing study of causal mechanisms and development of novel therapeutic interventions.

5/6th联合切除术，高盐饮食和一氧化氮合酶抑制剂诱导Lewis大鼠慢性肾脏病

Chronic kidney disease (CKD) is a global problem. Slowing CKD progression is a major health priority. Since CKD is characterized by complex derangements ...

A Murine Model of Irreversible and Reversible Unilateral Ureteric Obstruction

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. Presented here is a model of obstructive nephropathy induced by unilateral ureteric obstruction (UUO), which can either be irreversible (UUO) or reversible (R-UUO). In the irreversible UUO model, the ureter may be obstructed for variable periods of time in order to induce increasingly severe renal inflammation and interstitial fibrotic scarring. In the reversible R-UUO model the ureter is obstructed to induce hydronephrosis, tubular dilation and inflammation. After a suitable period of time the ureteric obstruction is then surgically reversed by anastomosis of the severed previously obstructed ureter to the bladder in order to allow complete decompression of the kidney and restoration of urinary flow to the bladder. The irreversible UUO model has been used to investigate various aspects of renal inflammation and scarring including the pathogenesis of disease and the testing of potential anti-inflammatory or anti-fibrotic therapies. The more challenging model of R-UUO has been used by some investigators and does offer significant research potential as it allows the study of inflammatory and immune processes and tissue remodeling in an injured and scarred kidney following the removal of the injurious stimulus. As a result, the R-UUO model offers investigators the opportunity to explore the resolution of kidney inflammation together with key aspects of tissue repair. These experimental models are of relevance to human disease as patients often present with obstruction of the renal tract that requires decompression and are commonly left with significant residual kidney impairment that has no current treatment options and may lead to eventual end stage kidney failure.

The murine model of irreversible unilateral ureteric obstruction (UUO) is presented together with the model of reversible UUO in which the ureteric obstruction is relieved by anastomosis of the severed ureter into the bladder. These models enable the study of renal inflammation and scarring as well as tissue remodeling.

的不可逆和可逆单侧输尿管梗阻小鼠模型

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. Presented here is a model of obstructive ...

A Modified Two Kidney One Clip Mouse Model of Renin Regulation in Renal Artery Stenosis

Renal artery stenosis is a common condition in patients with coronary or peripheral vascular disease where the renin angiotensin aldosterone system (RAAS) is overactivated. In this context, there is a narrowing of the renal arteries that stimulate an increase in the expression and release of renin, the rate-limiting protease in RAAS. The resulting rise in renin expression is a known driver of renovascular hypertension, frequently associated with kidney injury and end organ damage. Thus, there is a great interest in developing novel treatments for this condition. The molecular and cellular mechanism of renin control in renal artery stenosis is not fully understood and warrants further investigation. To induce renal artery stenosis in mice, a modified 2 kidney 1 clip (2K1C) Goldblatt mouse model was developed. The right kidney was stenosed in wild type mice and sham operated mice were used as control. After renal artery stenosis, we determined renin expression and kidney injury. Kidneys were harvested, and fresh cortices were used to determine protein and mRNA expression of renin. This animal model is reproducible and can be used to study pathophysiological responses, molecular and cellular pathways involved in renovascular hypertension and kidney injury.

A modified 2 kidney 1 clip (2K1C) Goldblatt mouse model was developed using polyurethane tubing to initiate renal artery stenosis, inducing an increase in renin expression and kidney injury. Here, we describe a detailed procedure of preparing and placing the cuff onto the renal artery to generate a reproducible and consistent 2K1C mouse model.

肾动脉狭窄肾素调节的改良双肾单夹小鼠模型

Renal artery stenosis is a common condition in patients with coronary or peripheral vascular disease where the renin angiotensin aldosterone system (RAAS) ...

An Effective Mouse Model of Unilateral Renal Ischemia-Reperfusion Injury

Ischemia-reperfusion injury (IRI) is the leading cause of acute renal failure and is a significant contributor to delayed graft function. Animal models are the only available resources that mimic the complexities of the IRI-associated damage encountered in vivo. This paper describes an effective mouse model of unilateral renal IRI that delivers highly reproducible data. Ischemia is induced by occluding the right renal pedicle for 30 min followed by reperfusion. In addition to the surgical procedure, a sequential overview of the expected physiological and histopathological changes following renal IRI will be provided by comparing data from seven different reperfusion times (4 h, 8 h, 16 h, 1 day, 2 days, 4 days, and 7 days). Critical data for planning experiments ahead, such as mean surgical time, average anesthetic consumption, and body weight changes over time, will be shared. This work will help researchers implement a reliable renal IRI model and select the appropriate reperfusion time that aligns with their intended investigative goals.

Renal ischemia-reperfusion injury is associated with high morbidity and mortality in hospitalized patients. Here, we present a simple and effective mouse model of unilateral renal ischemia-reperfusion injury and provide a sequential overview of representative pathological changes observed in the kidney.

单侧肾缺血再灌注损伤的有效小鼠模型

Ischemia-reperfusion injury (IRI) is the leading cause of acute renal failure and is a significant contributor to delayed graft function. Animal models ...

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 ...

<meta charset="utf8"></head><body>高脂饮食和 STZ 注射治疗糖尿病心肌病的小鼠模型

Modeling and Evaluation of Murine Diabetic Cardiomyopathy Model

The underlying pathophysiological mechanisms of diabetic cardiomyopathy (DbCM), a leading cause of mortality among patients with type 2 diabetes mellitus (T2DM), remain poorly understood. The myocardial toxicity associated with T2DM is attributed to factors such as lipotoxicity, glucotoxicity, oxidative stress, reduced cardiac efficiency, and lipoapoptosis. Compared to rats, mice offer greater accessibility, cost-effectiveness, and broader applicability for animal experiments. Insulin resistance and impaired insulin secretion are crucial factors in the pathophysiology of T2DM. We introduce a novel nongenetic murine model that replicates the progression of human DbCM induced by a combination of high-fat diet (HFD) feeding and streptozotocin (STZ) injection. In this study, we used wild-type C57BL/6J mice, administering an HFD regimen for 12 weeks, followed by intraperitoneal injections of STZ for an additional 12 weeks to induce characteristic manifestations of T2DM. We conducted oral glucose tolerance tests and measured serum insulin concentrations to confirm the development of insulin resistance and insufficient insulin secretion. Cardiac structure and function were rigorously assessed through noninvasive transthoracic echocardiography. Pathological characteristics were evaluated through Masson's trichrome staining and wheat germ agglutinin (WGA) staining, revealing pathological features related to DbCM. Therefore, we provide a robust and versatile method for establishing a nongenetic murine model of DbCM.

<meta charset="utf8"></head><body>小鼠糖尿病心肌病模型的建模与评价

This protocol outlines a method for inducing diabetic cardiomyopathy through a combination of high-fat diet feeding and streptozotocin injection. This approach aims to provide a reliable framework for scientific investigation into diabetic cardiomyopathy and to explore potential avenues for clinical treatment applications.

小鼠糖尿病心肌病模型的建模与评价

The underlying pathophysiological mechanisms of diabetic cardiomyopathy (DbCM), a leading cause of mortality among patients with type 2 diabetes mellitus ...

一种诱导实验性尿毒症心肌病的小鼠 5/6^th肾切除模型

摘要

探索更多视频

此视频中的章节

常温心脏骤停和心肺复苏术：缺血再灌注损伤的小鼠模型

小鼠腹膜透析模型导管置入及5/6 切除术的手术技术

小鼠肾中微RNA表达与单体尿阻的定量实时PCR评价

5/6th联合切除术，高盐饮食和一氧化氮合酶抑制剂诱导Lewis大鼠慢性肾脏病

的不可逆和可逆单侧输尿管梗阻小鼠模型

肾动脉狭窄肾素调节的改良双肾单夹小鼠模型

单侧肾缺血再灌注损伤的有效小鼠模型

血液透析通路相关手功能障碍的小鼠模型

小鼠糖尿病心肌病模型的建模与评价

小鼠糖尿病心肌病模型的建模与评价