Name: an experimental model of diet-induced metabolic syndrome in rabbit: methodological considerations, development, and assessment
Uploaded: 2018-04-20T13:00:00.000+00:00
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Description: Watch this Scientific Journal Video about An Experimental Model of Diet-Induced Metabolic Syndrome in Rabbit: Methodological Considerations, Development, and Assessment at JoVE.com

这项工作得到了加泰罗尼亚自治区 Valenciana (GV2015-062)、大学 de València (UV-INV-PRECOMP14-206372) 对、加泰罗尼亚自治区 Valenciana (PROMETEOII/2014/037) 和干杯 III. 菲德基金 (CIBERCV CB16/11/0486) FJC 的支持。

这项研究中使用的动物护理和实验性议定书符合欧盟关于保护用于科学目的的动物的2010/63 号指令, 并得到了机构动物保育和使用委员会 (2015/变现/豌豆/00049) 的批准。
注: 该议定书包括长期管理高脂肪、高蔗糖饮食28周, 以及对大都会主要成分的评估。我们使用11成年男性新西兰白 (NZW) 兔子重 4.39, 0.14 (s.d.) 公斤, 是 20-22 星期老在实验性协议初。他们被安置在一个房间与湿气 (50 5%) 和温度 (20 和1.5 °c) 被控制的情况以12小时光周期。"周" 和 "饮食" 一词可在协议步骤中交替使用。
1. 饮食管理
<ol><li>
获取或准备饮食
	<ol>
<li>获得一个商业上可用的高脂肪饮食加氢化椰子油 (10%) 和猪油 (5%)19。这种饮食将提供 3.7 kcal·g-1。</li>
		<li>通过在灭菌水中溶解适量的蔗糖 (例如), 准备5到 15%蔗糖溶液, 用300克蔗糖在2升的库存中使用15% 蔗糖溶液。15% 解决方案将提供 0.6 kcal·mL-1。</li>
		<li>获取可商业化使用的控制膳食19, 该控件提供 2.7 kcal·g-1。</li>
	</ol>
</li>
	<li>
对动物进行4周的驯化
	<ol>
<li>每天喂养控制组120克控制饮食的动物。提供水ad 随意。</li>
		<li>饲料动物在大都会集团 250 g 周开始与50% 控制和50% 高脂肪的周, 逐步增加到100% 高脂肪的周结束4。 注: 目标是实现: (i) 35% 控制和65% 高脂肪周年底 1;(ii) 25% 管制及75% 个高脂肪的周至2底; 及(iii) 15% 控制及85% 高脂肪周至3底。(iv) 在4周底前, 100% 个高脂食物。</li>
		<li>在一开始, 给在大都会组水中的动物以5% 蔗糖, 并在4第周结束时将蔗糖浓度提高到 15%.</li>
		<li>登记每日摄入的周和蔗糖溶液, 以计算热量摄入量, 根据1.1.1 提供的价值。和1.1.2。</li>
	</ol>
</li>
	<li>
诱导的大都会 (28 周)
	<ol>
<li>饲料控制组120克控制周和水的每一个动物每日随意。</li>
		<li>喂动物在大都会组250克高脂肪周和15% 蔗糖在水中。每三天更换一次日、蔗糖溶液。</li>
		<li>每日权衡剩余的周和水, 以估计每日摄入量。</li>
	</ol>
</li>
</ol>2. 形态学评估
<ol><li>每周测量动物的体重。</li>
	<li>测量高度,长度,腹部轮廓, 和胫骨长度, 并估计BMI在实验性饮食的管理和14周和28的麻醉动物。<ol>
<li>Cannulate 右耳边缘静脉与无菌一次性导管 (18-22 克) 和注射异丙酚 (8 mgkg-1), 其次是1.5 毫升的 0.9% NaCl 溶液。在麻醉家兔中, 执行后续步骤中列出的测量结果。</li>
		<li>测量高度和长度。使用测量带, 测量和记录的距离, 从鼻子到脚跟在侧压压位置 (长度)。在同一位置, 采取从肩的距离从肩膀到爪子的尖端 (高度)。</li>
		<li>计算身体质量指数 (BMI)20体重 (千克) ·[车身长度 (m) x 高度 (m)]-1。</li>
		<li>在腹部轮廓周围轻轻地放置测量带, 并在仰卧位置与动物进行测量。</li>
		<li>从膝关节下部到跟腱的插入测量胫骨长度。</li>
	</ol>
</li>
</ol>3. 空腹血糖和静脉葡萄糖耐受试验 (IVGTT)
注意: 建议在一天中的同一时间 (即, 2-下午3点) 启动过程。
<ol><li>用60克葡萄糖在100毫升0.9% 氯化钠溶液中制备葡萄糖库存溶液 (60%)。</li>
	<li>快速动物为 7 h (去除食物和维护水), 然后安置神志清楚的兔子在抑制剂在容易的位置。准备葡萄糖表 (插入一个新的条到米), 并采取第一个样本从左耳朵边缘静脉使用柳叶刀, 以获得一滴血。然后用葡萄糖表对血滴进行接触, 用血糖计测定血糖水平, 以测定空腹血糖。</li>
	<li>Cannulate 右耳边缘静脉与一次性导管 (18-22 克) 和注射一个60% 葡萄糖溶液丸 (0.6 g·kg-1)。 注: 准备丸, 加入1毫升/公斤的葡萄糖储存。</li>
	<li>在葡萄糖注射后, 用柳叶刀 (一滴血) 在15、30、60、90、120和180分钟取血样, 并用葡萄糖计与3.2 进行分析。</li>
	<li>取出一次性导管, 用纱布捏住导管插入部位。一旦血液凝结, 取出纱布, 检查动物的状态。</li>
</ol>4. 血压
<ol><li>准备采集系统, 包括压力传感器, 10 毫升注射器, 0.9% 氯化钠, 三路旋塞阀, 放大器, 和电脑/笔记本电脑与采集软件 (为血压记录)。</li>
	<li>设置设备。首先, 将三路旋塞阀和注射器放在压力传感器、传感器和导管之间, 并将压力传感器连接到放大器。然后将放大器连接到 PC/笔记本电脑。</li>
	<li>根据制造商的建议, 执行压力传感器校准。</li>
	<li>把有意识的动物放在兔子抑制剂的俯卧位置。在插管前预热耳朵, 然后局部应用局部麻醉 (2.5% 利多卡因/乳) 在耳周围的插入部位。轻轻地敲击血管包裹运行的区域, 以方便地识别动脉。然后在左耳中央动脉插入一根无菌导管 (18-22 克)。松开限制, 让动物保持安静30分钟。</li>
	<li>记录血压连续20分钟直接从动脉导管, 放置压力传感器放置在动物旁边的心脏水平 (采样频率: 1 赫, 请参见图 5B)。 注: 为了保持血压 (bp) 记录不受凝血干扰 (bp 信号失去振幅或消失), 应进行氯化钠 (0.9%) 注射液。使用三路旋塞阀, 关闭从传感器到导管的电路, 打开从注射器到导管的电路, 并注入 1-2 毫升。这将清除导管中可能形成的血块。然后, 打开传感器和导管之间的电路, 并在信号恢复后继续录音。</li>
	<li>录音完成后, 取出导管, 用纱布夹在导管插入部位, 以防止失血。一旦血液凝结, 取出纱布, 检查动物的状态。</li>
</ol>5. 等离子测量
注意: 建议在一天中的同一时间 (即, 2-下午3点) 启动过程。
<ol><li>快速动物为 7 h (去除食物和维护水), 然后安置神志清楚的动物在抑制剂在容易的位置和插入无菌 21 G 针在左耳边缘静脉。一旦血液开始滴水, 丢弃第一滴, 并收集血液样本的 EDTA 管高达水平表明在管。把样品储存在冰上。</li>
	<li>离心血液样品在 1500 x g, 15 分钟, 4 °c。离心后, 用吸管吸入等离子, 并准备整除数250µL。</li>
	<li>立即分析新鲜样品。基本控制参数如下: 甘油三酯, 总胆固醇, HDL 和 LDL 胆固醇。 注: 未新鲜分析的样品应立即存放在-80 摄氏度冷藏库中。如果有兴趣对血浆样品中的血糖进行分析, 血糖测试应使用草酸盐管代替 EDTA。</li>
</ol>

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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Age-specific+prevalence+of+the+metabolic+syndrome+defined+by+the+International+Diabetes+Federation+and+the+National+Cholesterol+Education+Program:+The+Norwegian+HUNT+2+study.">Age-specific prevalence of the metabolic syndrome defined by the International Diabetes Federation and the National Cholesterol Education Program: The Norwegian HUNT 2 study.</a> BMC Public Health. 7, 220 (2007).</li></ol>

作者声明他们没有竞争的财政利益。

建立合适的实验模型, 可以为研究肿瘤的发展提供更一致、更可靠的方法, 同时也需要了解器官和系统重塑的基本机制。在这里, 我们描述了一些方法来建立一个相关的饮食诱导大鼠群的实验模型, 以及如何评估这一组代谢和心血管异常的主要组成部分, 这一模式的特点: 中央肥胖, 高血压,葡萄糖不耐受, 血脂异常与低 HDL, 高 LDL, 和 TG 水平的增加。
该模型的一个主要力量是能够研究的条件, 在临床表现之前的病理。事实上, 关于新陈代谢的变化, 在28周内, 动物没有发展 II 型糖尿病并且处于糖尿病前期状态 (图4)。同样, 血浆生化标志物在脂质剖面上明显改变, LDL 和 TG 增加, HDL 减少, 但总胆固醇 (表 3) 没有变化, 这是动脉粥样硬化发展的关键因素。尽管我们可以观察到28周 (图 5) 的收缩、舒张和图的增加, 但这可以被认为是轻度高血压。总的来说, 这些代谢和心血管指标的影响是适度的, 但这种模式可以使研究的状态之前的表现 (和在大多数情况下是不可逆转的) 病理, 从而使鉴定的临床前和临床标记, 可能使检测的病人处于危险中。
此外, 不像其他大都会动物模型 (老鼠, 鼠, 狗), 自发或转基因兔模型可以开发的所有组成部分的大都会。有趣的是, 据报道, 大都会的不同成分组合可以放大心血管风险。事实上, 当更多的组组出现23时, 高血压产生的病理重塑就会加重。该实验模型可以对底层机理进行研究, 并对不同组分的影响进行组合。此外, 鉴于饮食影响全身新陈代谢, 使用饮食诱导模型具有重要的意义, 密切效仿发生在人类大都会的19。
最后, 但不是最不重要的力量, 是平衡在翻译研究和经济费用之间的相关性和影响。一方面, 我们发现猪模型与人类非常相似, 但在时间、资源和经济成本方面却非常昂贵。另一方面, 我们有啮齿类动物模型, 它易于实现, 成本很少, 但泛化能力较低。兔子模型代表的中间点, 因为它是足够灵活的许多不同类型的研究, 同时避免一些缺陷的大型动物模型, 并显示类似的血流动力学和神经体液变化观察到在人类大都会 8, 10,19。
应考虑以下所述方法的局限性。关于中央肥胖和体脂分布, 磁共振成像的使用将是金标准, 如果有的话, 否则使用28周结束时内脏脂肪的量化。其他用于纵向研究的非侵入性方法, 如 X 射线计算机断层扫描, 将更适当的24。我们测量了腹部周长和 BMI (表 2), 这也被用于一些研究, 在兔子作为衡量中央肥胖25,11,26。胫骨长度测量也可以更精确使用声像图或腿部摄影。为了确定这种慢性模型中葡萄糖不耐受的原因是胰岛素抵抗, 还是胰岛素分泌减少, 应使用胰岛素耐受试验或确定空腹胰岛素水平来确定胰岛素抵抗。
最后, 为了改进该模型, 可以采取一些措施。我们可能已经获得了更快的增长, 空腹的结合, 短暂的四氧嘧啶注射液和高脂肪, 高蔗糖饮食, 但随后的表型不能仅归因于饮食。年龄可能也扮演一个重要角色, 因为我们与幼小成年兔子合作 (4.5 月大当动物到达动物设施, 12.5-13 月大在实验性协议的末端) 和大都会经常发生在更旧的年龄27。不幸的是, 年长的兔子没有商业化的供应。这将是有趣的测试这个模型在老动物和观察如果表型是否加重。
本文的研究方法为实验兔大鼠肿瘤模型的发展提供了一个有价值的工具, 旨在阐明大都会的病理重塑在不同器官和系统, 并获得对这个复杂的病理学的理解。最后, 由于 NZW 兔子是久坐的动物, 这种饮食诱导模型可以用来研究不同的病理成分如何进化的方式比在人类大都会发生, 并可以让新的观点, 以了解病理生理学机制参与的疾病的进展, 鉴定的前体和临床标志, 以确定患者的风险, 甚至测试新的治疗方法, 治疗这种复杂的病理。

由于久坐的生活方式和不健康的饮食习惯的增加, 肥胖和代谢综合征 (大都会) 已经成为公共卫生和临床实践的一个日益严重的问题, 因为1。有几个大都会的定义, 但他们大多描述它作为一个集群的心血管和新陈代谢的变化, 如腹部肥胖, 降低高密度脂蛋白和高 LDL 胆固醇, 甘油三酯, 葡萄糖不耐受和高血压2 ,3,4。诊断要求这五标准中有三是存在的。
由于动物模型, 基础研究已经能够调查的机制, 如病理过程, 如大都会。已经使用了几种动物模型, 但最重要的是, 选择模型再现人类病理学的主要临床表现 (图 1)。有了这个目的, 被认为类似人类的动物模型, 主要是犬和猪, 已经开发 (见 Verkest5和张 & Lerman6进行审查)。然而, 由于狗的动脉粥样硬化或高血糖的发展, 犬的模型不显示所有的组成部分, 由于饮食是可疑的5。猪模型呈现出与人类最具解剖学和生理上的相似性, 从而为阐明大鼠的机制提供了重要的预测能力, 但其维护和实验过程的复杂性使得使用这个模型非常劳力密集和昂贵的6。
另一方面, 鼠类模型 (鼠和鼠), 饮食诱导自发和转基因, 已广泛用于研究肥胖, 高血压和大都会, 其病理结果在不同的器官和系统 (见黄et 。7进行审阅)。虽然这些模型的使用比狗或猪更实惠, 但它们有重要的缺点。事实上, 根据菌株的不同, 动物会发展出一些组, 而其他如高血压、高血糖和 hyperinsulinemia 则缺席7。此外, 大都会的主要成分之一, 肥胖, 在一些转基因菌株, 不仅取决于与饮食相关的因素, 而是表明, 一些动物变得肥胖与正常或甚至减少食物摄入量8。最后, 小鼠和大鼠显示胆固醇酯转运蛋白 (CETP) 的自然缺陷, 并以高密度脂蛋白为主要的胆固醇转运手段, 使其对动脉粥样硬化的发展具有相对的抵抗力。这是一个重要的区别, 脂代谢与人, 谁表达 CETP 和运输他们的胆固醇主要在 LDL9。
相反, 实验兔子代表了大动物和啮齿动物实验模型之间的中间阶段。因此, 兔子可以很容易地提交到不同类型的协议, 对人员和维护的最低要求, 比较大的动物模型更容易在实验过程中处理。此外, 据报道, 喂养高脂肪饮食的兔子有类似的血流动力学和神经体液的变化, 肥胖的人 8, 10, 11.注意, 关于脂质代谢, 兔子有丰富的 CETP 在血浆和他们的脂蛋白配置文件是 LDL 丰富的12, 这也类似于人类。此外, 兔子非常迅速地发展高脂血症, 因为作为食草动物, 他们非常敏感的膳食脂肪13。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/57117/57117fig1.jpg"> 图 1: 对大都会动物模型的比较.参见 Verkest5、张和 Lerman6和黄et 等。7进行审阅。"" 表示优势, "" 表示劣势.<img alt="Equation 1" src="/files/ftp_upload/57117/57117eq1.jpg"><img alt="Equation 2" src="/files/ftp_upload/57117/57117eq2.jpg">*有争议, 取决于研究, 如卡罗尔等所指出的**。8, 一些转基因菌株在食物摄入量之外就变得肥胖。化学合成: 胆固醇酯转运蛋白。GTT: 葡萄糖耐受性试验。<a href="//cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/57117/57117fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
为了阐明大鼠在不同器官和系统中产生的病理重塑的基本机制, 并了解这一复杂的病理学, 选择一个重现主要成分的实验模型。人类大都会是必不可少的。由于它与人类生理的相似性以及在慢性协议和测量中的使用负担能力, 兔子可以提供许多优势。在这条线, 很少有饮食诱导的兔子模型使用高脂肪和高蔗糖饮食已使用14,15、16、17、18、19 (表 1) 和一个在与器官重塑有关的表型时, 对大都会不同成分的表征具有重要意义。因此, 本文的主要目的是描述建立饮食诱导的大鼠大鼠模型的方法, 允许研究其病理生理学和对器官重塑的影响。
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody>
<tr>
<td rowspan="2">研究</td>
			<td rowspan="2">饮食</td>
			<td rowspan="2">时间</td>
			<td rowspan="2">品种</td>
			<td colspan="4">大都会组件</td>
		</tr>
<tr>
<td>Ob</td>
			<td>HT</td>
			<td>汞</td>
			<td>Dl</td>
		</tr>
<tr>
<td rowspan="2">尹等 (2002)14</td>
			<td>·   10% 脂肪</td>
			<td rowspan="2">24周</td>
			<td>·     男 NZW</td>
			<td rowspan="2"><img alt="Equation 2" src="/files/ftp_upload/57117/57117eq2.jpg"></td>
			<td rowspan="2">-</td>
			<td rowspan="2"><img alt="Equation 1" src="/files/ftp_upload/57117/57117eq1.jpg"></td>
			<td rowspan="2"><img alt="Equation 1" src="/files/ftp_upload/57117/57117eq1.jpg"></td>
		</tr>
<tr>
<td>·   37% 蔗糖</td>
			<td>·     2公斤</td>
		</tr>
<tr>
<td rowspan="2">赵等 (2007)15</td>
			<td>·   10% 脂肪</td>
			<td rowspan="2">36周</td>
			<td>·     马累 JW</td>
			<td rowspan="2"><img alt="Equation 1" src="/files/ftp_upload/57117/57117eq1.jpg"></td>
			<td rowspan="2"><img alt="Equation 2" src="/files/ftp_upload/57117/57117eq2.jpg"></td>
			<td rowspan="2"><img alt="Equation 2" src="/files/ftp_upload/57117/57117eq2.jpg"></td>
			<td rowspan="2"><img alt="Equation 2" src="/files/ftp_upload/57117/57117eq2.jpg"></td>
		</tr>
<tr>
<td>·   30% 蔗糖</td>
			<td>·     16周</td>
		</tr>
<tr>
<td rowspan="3">Helfestein 等 (2011)16</td>
			<td>·   10% 脂肪</td>
			<td rowspan="3">24周</td>
			<td>·     男 NZW</td>
			<td rowspan="3"><img alt="Equation 2" src="/files/ftp_upload/57117/57117eq2.jpg"></td>
			<td rowspan="3">-</td>
			<td rowspan="3"><img alt="Equation 1" src="/files/ftp_upload/57117/57117eq1.jpg"></td>
			<td rowspan="3"><img alt="Equation 1" src="/files/ftp_upload/57117/57117eq1.jpg"></td>
		</tr>
<tr>
<td>·   40% 蔗糖</td>
			<td>·     12周</td>
		</tr>
<tr>
<td>·   0.5-0.1 胆固醇</td>
			<td></td>
		</tr>
<tr>
<td rowspan="2">宁等 (2015)17</td>
			<td>·   10% 脂肪</td>
			<td rowspan="2">8-16 周</td>
			<td>·     男 WHHL</td>
			<td rowspan="2"><img alt="Equation 2" src="/files/ftp_upload/57117/57117eq2.jpg"></td>
			<td rowspan="2">-</td>
			<td rowspan="2"><img alt="Equation 2" src="/files/ftp_upload/57117/57117eq2.jpg"></td>
			<td rowspan="2"><img alt="Equation 1" src="/files/ftp_upload/57117/57117eq1.jpg"></td>
		</tr>
<tr>
<td>·   30% 果糖 *</td>
			<td>·     12周</td>
		</tr>
<tr>
<td rowspan="2">刘等 (2016)18</td>
			<td>·   10% 脂肪</td>
			<td rowspan="2">48周</td>
			<td>·     男 NZW</td>
			<td rowspan="2"><img alt="Equation 2" src="/files/ftp_upload/57117/57117eq2.jpg"></td>
			<td rowspan="2">-</td>
			<td rowspan="2"><img alt="Equation 1" src="/files/ftp_upload/57117/57117eq1.jpg"></td>
			<td rowspan="2"><img alt="Equation 1" src="/files/ftp_upload/57117/57117eq1.jpg"></td>
		</tr>
<tr>
<td>·   30% 蔗糖</td>
			<td>·     12周</td>
		</tr>
<tr>
<td>咏叹调-Mutis 等 (2017)19</td>
			<td>·   15% 脂肪</td>
			<td>28周</td>
			<td>·     男 NZW</td>
			<td><img alt="Equation 1" src="/files/ftp_upload/57117/57117eq1.jpg"></td>
			<td><img alt="Equation 1" src="/files/ftp_upload/57117/57117eq1.jpg"></td>
			<td><img alt="Equation 1" src="/files/ftp_upload/57117/57117eq1.jpg"></td>
			<td><img alt="Equation 1" src="/files/ftp_upload/57117/57117eq1.jpg"></td>
		</tr>
</tbody></table>表 1: 采用高脂高蔗糖膳食诱导的大鼠兔模型.符号 "" 指示缺勤、"" 存在和 "-" 未计算.<img alt="Equation 2" src="/files/ftp_upload/57117/57117eq2.jpg"><img alt="Equation 1" src="/files/ftp_upload/57117/57117eq1.jpg">限制.WHHL, 渡边遗传性 hiperlipidemic 兔。JW, 日本白兔。肥胖症高血压汞, 高血糖。Dl, 血脂异常。

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http://www.bt-ingenieros.com/fluidos-y-combustibles/961-juego-de-4-embudos-de-plastico.html?gclid=EAIaIQobChMIuInui_y-1QIVASjTCh28Zwf-EAQYBSABEgK7xPD_BwE</td></tr><tr><td>Introcan Certo 22G blue</td><td>B Braun</td><td>4251318</td><td>Peripheral intravenous catheter&lt;br/&gt; http://www.bbraun-vetcare.es/producto/introcan-</td></tr><tr><td>Propofol Lipuro 10 mg/ml vial 20 ml</td><td>B Braun</td><td>3544761VET</td><td>General intravenous anesthetic&lt;br/&gt; http://www.bbraun-vetcare.es/producto/propofol-lipuro-1-</td></tr><tr><td>FisioVet serum solution 500ml</td><td>B Braun</td><td>472779</td><td>Scale&lt;br/&gt; https://www.soehnle-professional.com/en/productgroup/details/103/veterinary-scale</td></tr><tr><td>Askina Film Vet 1,25cm x 5m</td><td>B Braun</td><td>OCT13501</td><td>Plastic Plaster&lt;br/&gt; http://www.bbraun-vetcare.es/producto/askina-film-vet</td></tr><tr><td>Askina Film Vet 2,50cm x 5m</td><td>B Braun</td><td>OCT13502</td><td>Plastic Plaster&lt;br/&gt; http://www.bbraun-vetcare.es/producto/askina-film-vet</td></tr><tr><td>Injekt siringe 10ml luer</td><td>B Braun</td><td>4606108V</td><td>Injection-aspiration syringe of two single-use bodies&lt;br/&gt; http://www.bbraun-vetcare.es/producto/injekt-</td></tr><tr><td>Seca 201</td><td>seca</td><td>seca 201</td><td>Ergonomic tape for measuring perimeters&lt;br/&gt; https://www.seca.com/es_es/productos/todos-los-productos/detalles-del-producto/seca201.html#referred</td></tr><tr><td>Sterican 21Gx1&quot; - 0,8x25mm verde</td><td>B Braun</td><td>4657543</td><td>Single Use Hypodermic Needle&lt;br/&gt; http://www.bbraun-vetcare.es/producto/agujas-hipodermicas-sterican-</td></tr><tr><td>CONTOURNEXT-Meter</td><td>BAYER</td><td>84413470</td><td>Blood glucose analysis system&lt;br/&gt; http://www.contournextstore.com/en/contour-next-meter-2</td></tr><tr><td>CONTOUR NEXT test strips</td><td>BAYER</td><td>83624788</td><td>Blood glucose test strips&lt;br/&gt; http://www.contournextstore.com/en/contour-next-test-strips-100-ct-package</td></tr><tr><td>MICROLET NEXT LANCING DEVICE</td><td>BAYER</td><td>6702</td><td>Lancing device&lt;br/&gt; http://www.contournextstore.com/en/new-microlet-next-lancing-device</td></tr><tr><td>MICROLET 2 Colored Lancets</td><td>BAYER</td><td>81264857</td><td>Ultra-thin sterile lancet for capillary puncture&lt;br/&gt; http://www.contournextstore.com/en/microlet2-colored-lancets-100s</td></tr><tr><td>Injekt 20ml luer siringe</td><td>B Braun</td><td>4606205V</td><td>Scale&lt;br/&gt; https://www.soehnle-professional.com/en/productgroup/details/103/veterinary-scale</td></tr><tr><td>Askina Mullkompressen 7,5x7,5cm - sterile</td><td>B Braun</td><td>9031219N</td><td>Sterile gauze packets in envelopes&lt;br/&gt; http://www.bbraun-vetcare.es/producto/askina-mullkompressen-esteril</td></tr><tr><td>Emla lidocaine/prilocaine</td><td>AstraZeneca</td><td>not available</td><td>Local anesthetics&lt;br/&gt; https://www.astrazeneca.es/areas-terapeuticas/neurociencias.html</td></tr><tr><td>Introcan Certo 18G short</td><td>B Braun</td><td>4251342</td><td>Peripheral intravenous catheter&lt;br/&gt; http://www.bbraun-vetcare.es/producto/introcan-</td></tr><tr><td>Introcan Certo 20G</td><td>B Braun</td><td>4251326</td><td>Peripheral intravenous catheter&lt;br/&gt; http://www.bbraun-vetcare.es/producto/introcan-</td></tr><tr><td>Blood Pressure Transducers-MA1 72-4497</td><td>Harvard Apparatus</td><td>724497</td><td>Transducer for monitoring blood pressure&lt;br/&gt; http://www.harvardapparatus.com/physiology/physiological-measurements/transducers/pressure-transducers/research-grade-pressure-transducers.html</td></tr><tr><td>PowerLab 2/26</td><td>AD Instruments</td><td>ML826</td><td>Amplifier&lt;br/&gt; https://www.adinstruments.com/products/powerlab</td></tr><tr><td>LabChart ver. 6</td><td>AD Instruments</td><td>not available</td><td>Acquisition software&lt;br/&gt; https://www.adinstruments.com/products/labchart</td></tr><tr><td>Animal Bio Amp</td><td>AD Instruments</td><td>FE136</td><td>Amplifier&lt;br/&gt; https://www.adinstruments.com/products/bio-amps#product-FE136</td></tr><tr><td>K2EDTA 7.2mg</td><td>BD</td><td>367861</td><td>Blood collection tubes&lt;br/&gt; http://catalog.bd.com/nexus-ecat/getProductDetail?productId=367861</td></tr><tr><td>Centrifuge</td><td>SciQuip</td><td>2-16KL</td><td>Centrifuge&lt;br/&gt; http://www.sigma-centrifuges.co.uk/store/products/refrigerated-sigma-2-16k-centrifuge/</td></tr><tr><td>Eppendorf Reference 2, 100 &amp;ndash; 1000 &amp;mu;L</td><td>Eppendorf</td><td>4920000083</td><td>Pipette&lt;br/&gt; https://online-shop.eppendorf.es/ES-es/Pipeteo-44563/Pipetas-44564/Eppendorf-Reference2-PF-42806.html</td></tr><tr><td>Eppendorf Safe-Lock Tubes, 0.5 mL</td><td>Eppendorf</td><td>30121023</td><td>Tubes&lt;br/&gt; https://online-shop.eppendorf.es/ES-es/Puntas-tubos-y-placas-44512/Tubos-44515/Eppendorf-Safe-Lock-Tubes-PF-8863.html</td></tr><tr><td>NZW rabbits (16-18 weeks old)</td><td>Granja San Bernardo</td><td>not available</td><td>New Zealand White rabbits&lt;br/&gt; http://www.granjasanbernardo.com/en/welcome/</td></tr><tr><td>Sucrose&amp;nbsp;</td><td>Sigma</td><td>S0389-5KG</td><td>Sucrose for drinking solution&lt;br/&gt; http://www.sigmaaldrich.com/catalog/product/sigma/s0389?lang=es&amp;amp;region=ES</td></tr><tr><td>Rabbit maintenance control diet</td><td>Ssniff</td><td>V2333-000</td><td>Control diet&lt;br/&gt; http://www.ssniff.com/</td></tr><tr><td>Rabbit high-fat diet</td><td>Ssniff</td><td>S9052-E020</td><td>High-fat diet&lt;br/&gt; http://www.ssniff.com/</td></tr><tr><td>Rabbit rack and drinker</td><td>Sodispan</td><td>not available</td><td>Rack for rabbits&lt;br/&gt; https://www.sodispan.com/jaulas-y-racks/racks-conejo-y-cobaya/</td></tr><tr><td>Rabbit restrainer</td><td>Zoonlab</td><td>3045601</td><td>http://www.zoonlab.de/en/index.html</td></tr></tbody></table>

an experimental model of diet-induced metabolic syndrome in rabbit: methodological considerations, development, and assessment

近年来, 由于久坐的生活方式和不健康的饮食习惯的增加, 肥胖和代谢综合征已经成为公共卫生和临床实践的一个日益严重的问题。由于动物模型, 基础研究可以调查病理过程的机制, 如大都会。在这里, 我们描述的方法, 以建立实验兔模型的饮食诱导的大都会和它的评估。经过一段时间的驯化, 动物饲养高脂肪 (10% 氢化椰子油和5% 猪油), 高蔗糖 (15% 蔗糖溶于水) 饮食28周。在此期间, 进行了几项实验程序, 以评估各组的不同组成部分: 形态学和血压测量, 葡萄糖耐受性测定, 并分析了几种血浆标记物。在实验期结束时, 动物发育为中央性肥胖症、轻度高血压、前糖尿病和血脂异常, 低密度脂蛋白, 高 LDL, 并增加甘油三酯 (TG) 水平, 从而再现人类大都会的主要成分。这种慢性模型允许新的观点, 以了解疾病进展的基本机制, 检测的基础和临床标志, 使病人的识别风险, 甚至测试新的治疗治疗这种复杂病理的方法。

大都会代表一组代谢和心血管异常, 其研究可以通过使用实验模型来促进。事实上, 为了阐明由肿瘤组织产生的病理重塑的机制, 选择一个适当类似于人类状况和适合研究的实验模型是至关重要的。本文介绍了用饱和脂肪和蔗糖高的膳食诱导家兔大都会的方法, 并对其评价进行了详细的描述。使用饮食代替转基因动物模型是非常重要的, 因为饮食影响全身新陈代谢19, 因而类似于人类大都会发生的事情。我们使用了一个阶乘 (混合模型) 方差分析与两个因素, 一个重复的措施或 "内" 因素 (时间: 前, 周14和周 28, 取决于分析) 和一个 "之间的因素 (组: 控制和大都会) 的统计研究。当p < 0.05 时, 其重要性被接受。
高脂肪, 高蔗糖的饮食是很好的耐受动物。一个4周的驯化期是正确的过渡从以前的喂养制度, 以高脂肪, 高蔗糖饮食的必要。控制组中的动物喂养120克周, 这已被证明是适当的维护成年兔8。在实验性协议 (表 2) 结束之前, 大都会组中的兔子体重逐渐增加。动物每周要增加 50-100 克。重要的是, 兔子被单独安置在笼子里, 有足够的空间, 光线和环境浓缩 (图 2C), 并且每天对动物进行检查。此外, 每天都必须监督和登记饮食, 以达到体重增加和发现可能的健康问题, 因为兔子很容易被强调, 反应可能是停止营养消耗。此外, 由于高脂颗粒往往是非常不稳定的和容易失去一致性, 变成粉末, 兔子不吃, 这是至关重要的是准备的日常部分周非常仔细 (图 2A)。在图 3A中, 我们可以观察能量摄入的行为及其波动, 范围从250到815大卡。在图 3B中, 描述了不同能量源 (周和饮料) 的相对贡献。在14和28周有关键时期, 因为由于实验过程产生的压力, 兔子可能会减少周和水摄入量。每日量化允许快速识别这个问题, 可以避免通过引入控制周 (80% 高脂肪, 20% 控制) 和减少蔗糖溶液从15% 到 10%, 甚至 5%, 在2-3 天, 直到动物恢复正常摄取值。动物也发展了中央肥胖, 显示的体重增加, 腹部周长和 BMI (表 2), 这是密切相关的风险因素, 定义了中心3。
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/57117/57117fig2.jpg"> 图 2: 饮食管理.在面板A中, 对控制周 (顶部) 和高脂肪周 (以下) 进行了描述, 显示了由于添加的脂肪而导致两者之间的差异。为了避免使高脂肪颗粒不太可口的粉末, 有必要使用过滤器分离高脂肪颗粒粉 (面板 a, 底部)。在面板B中, 我们可以观察制作饮水解决方案所需的材料 (左), 以及如何在饮水机中分发库存解决方案的建议。最后, 动物的福利是非常重要的, 并且他们必须单独安置在笼子 (C) 以足够的空间和光, 并且, 如果可能, 环境充实 (例如, 平台, 玩具,等等)。<a href="https://cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/57117/57117fig2large.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/57117/57117fig3.jpg"> 图 3: 能量摄取量.实验期28周内每周摄取量的变化在对照组和大都会中被描述。在 b 组控制 (n = 5), 大都会 (n = 6) 中, 从高脂肪的食物中摄取的大卡的相对摄入量 (百分比) 和大都会动物的饮用溶液。误差线: SD. 从咏叹调-Mutis et修改。19<a href="https://cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/57117/57117fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody>
<tr>
<td></td>
			<td colspan="2">饮食前</td>
			<td colspan="2">14周</td>
			<td colspan="2">28周</td>
		</tr>
<tr>
<td></td>
			<td>控制</td>
			<td>纽约大都会</td>
			<td>控制</td>
			<td>纽约大都会</td>
			<td>控制</td>
			<td>纽约大都会</td>
		</tr>
<tr>
<td>重量 (千克)</td>
			<td>4.35 (0.15)</td>
			<td>4.43 (0.14)</td>
			<td>4.49 (0.12)</td>
			<td>5.42 (0.17)</td>
			<td>4.51 (0.13)</td>
			<td>5.75 (0.6)</td>
		</tr>
<tr>
<td>长度 (厘米)</td>
			<td>52.4 (1.6)</td>
			<td>53.6 (1.7)</td>
			<td>52.5 (0.8)</td>
			<td>54.4 (1.7)</td>
			<td>53.7 (0.7)</td>
			<td>54.6 (0.8)</td>
		</tr>
<tr>
<td>高度 (厘米)</td>
			<td>25.9 (0.7)</td>
			<td>25.5 (1.1)</td>
			<td>25.9 (2.2)</td>
			<td>26.1 (5.3)</td>
			<td>26.0 (1.0)</td>
			<td>26.1 (1.5)</td>
		</tr>
<tr>
<td>Abdom。周长 (厘米)</td>
			<td>39.8 (1.7)</td>
			<td>40.5 (1.4)</td>
			<td>38.5 (1.5)</td>
			<td>47.5 (2.2)</td>
			<td>38.1 (1.0)</td>
			<td>49.7 (3.5)</td>
		</tr>
<tr>
<td>胫骨长度 (厘米)</td>
			<td>16.4 (0.8)</td>
			<td>16.3 (0.7)</td>
			<td>16.7 (0.3)</td>
			<td>16.7 (0.4)</td>
			<td>17.4 (0.4)</td>
			<td>16.8 (0.6)</td>
		</tr>
<tr>
<td>BMI (千克/米2)</td>
			<td>32.8 (1.9)</td>
			<td>32.9 (2.6)</td>
			<td>32.8 (1.2)</td>
			<td>36.8 (1.9)</td>
			<td>32.6 (2.1)</td>
			<td>39.3 (6.0)</td>
		</tr>
</tbody></table>表 2: 形态学特征.我们在比较控制和对比时发现了差异. 大都会在星期14和28在重量 (主要作用 p = 0.003, η2 = 0.6; 配对比较在星期 14 p < 0.001 和星期 28 p < 0.001), 腹部周长 (主要作用 p < 0.001, η2 = 0.9 配对比较在星期 14 p < 0.001 和星期 28 p < 0.001) 和 BMI (主要作用 p = 0.016, η2 = 0.5; 配对比较在星期 14 p < 0.001 和星期 28 p < 0.001)。控制 (n = 5) 和大都会 (n = 6)。值表示为平均值 (SD)。从咏叹调-Mutis et al中修改。19。
关于空腹血糖, 对 IVGTT 的反应在葡萄糖稳态的表征中起关键作用21。我们观察到14周的轻度高血糖, 在28周 (图 4A), 它达到了一个高原并保持了类似的值。曲线下的区域 (联合自卫队) 也增加了在大都会组 (图 4B)。尽管根据空腹血糖来确定家兔 II 型糖尿病的截止值尚未被确认为19, 但有了这个实验性的协议, 兔子提交到28周高脂肪, 高蔗糖喂养发展前糖尿病随着空腹血糖和葡萄糖不耐受的减弱。
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/57117/57117fig4.jpg"> 图 4: 血糖调节.IVGTT 在14和28周的控制和大都会动物的结果显示在小组 A。在面板 B 中, 用盒形和晶须图描述了曲线下的面积 (哥伦比亚自卫队) 从0到180分钟的量化。这个参量在大都会动物增加了在星期14和28与控制 (主要作用 p = 0.001, η2 = 0.5; 配对比较在星期 14 p = 0.001 和星期 28 p = 0.002)。控制 (n = 5), 大都会 (n = 6)。从咏叹调-Mutis et al中修改。19<a href="https://cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/57117/57117fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
高血压与肥胖的严重程度有直接关系。喂养高脂肪、高蔗糖饮食28周的兔子显示, 14 周的收缩、舒张和平均血压都在增加, 血压在28周保持不变 (图 5C E)。鉴于血压和 BMI 的密切关系22, 这是非常重要的, 以确保动物体重递增, 以获得显著增加血压。
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/57117/57117fig5.jpg"> 图 5: 血压的修改.面板 A 描述插入在耳动脉中的导管。值得注意的是, 由于静脉和耳动脉在耳的牙列上非常紧密地贯穿, 所以区分它们是至关重要的。插管前, 最好是预热耳朵, 并在局部麻醉后, 轻轻拍打血管包裹的区域。动脉有一个较厚的血管壁和较浅的颜色比静脉, 和血液脉冲可以观察。小组 B 显示实验设置与压力传感器, 连接到放大器并且连续记录信号 (BP 录音)。在两个实验组中, C 和 D 面板显示14和28周的收缩和舒张压的盒子和胡须图。平均动脉压 (图) 显示在 E 组。我们在比较控制和对比时发现了差异. 大都会在星期14和28在收缩 (主要作用 p = 0.003, η2 = 0.4; 配对比较在星期 14 p = 0.029 和星期 28 p = 0.013), 舒张 (主要作用 p = 0.027, η2 = 0.3; 配对比较在星期 14 p = 0.036 和星期 28 p = 0.001) 和地图 (主要作用 p = 0.006, η2 = 0.4; 配对比较在星期 14 p = 0.027 和星期 28 p = 0.001)。控制 (n = 5), 大都会 (n = 6)。从咏叹调-Mutis et al中修改。19<a href="https://cloudflare-jove-com.bdigitaluss.remotexs.co/files/ftp_upload/57117/57117fig5large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
最后, 为了评估肿瘤的发展, 需要对血浆生化指标的变化进行评价。在这个慢性模型中, 我们观察到14周前脂质剖面的改变, 这一改变直到28周才保持稳定, 而没有进一步增加差异。血浆脂质的改变的特点是甘油三酯和 LDL 的增加, 高密度脂蛋白的减少, 以及在两个时间点 (周14和 28) 的控制下, 总胆固醇的变化与对照 (表 3)。
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody>
<tr>
<td></td>
			<td colspan="2">14周</td>
			<td colspan="2">28周</td>
		</tr>
<tr>
<td></td>
			<td>控制</td>
			<td>纽约大都会</td>
			<td>控制</td>
			<td>纽约大都会</td>
		</tr>
<tr>
<td>总胆固醇 (mg·dL-1)</td>
			<td>20.4 (2.3)</td>
			<td>24.0 (9.1)</td>
			<td>27.4 (15.7)</td>
			<td>21.2 (4.4)</td>
		</tr>
<tr>
<td>HDL (mg·dL-1)</td>
			<td>9.1 (4.2)</td>
			<td>4.3 (1.7)</td>
			<td>11.2 (4.2)</td>
			<td>5.1 (2.9)</td>
		</tr>
<tr>
<td>LDL (mg·dL-1)</td>
			<td>3.8 (1.1)</td>
			<td>8.7 (4.5)</td>
			<td>4.0 (1.2)</td>
			<td>13.8 (9.3)</td>
		</tr>
<tr>
<td>甘油三酯 (mg·dL-1)</td>
			<td>71.2 (58.8)</td>
			<td>118.0 (40.7)</td>
			<td>30.2 (11.4)</td>
			<td>76.8 (28.2)</td>
		</tr>
</tbody></table>表 3: 血浆生物化学的评估.在14和28的 HDL (主要效果 p = 0.008) 比较控制与大都会时, 我们发现了差异. η2 = 0.3; 在周 14 p = 0.006 和周 28 p = 0.037) 配对比较, LDL (主要作用 p = 0.040, η2 = 0.2; 配对比较在星期 14 p = 0.028和星期 28 p = 0.034) 和甘油三酯 (主要作用 p = 0.002, η2 = 0.4; 配对比较在星期 14 p = 0.004 和星期 28 p = 0.001)。控制 (n = 5) 和大都会 (n = 6)。值表示为平均值 (SD)。从咏叹调-Mutis et al中修改。19

Watch this Scientific Journal Video about An Experimental Model of Diet-Induced Metabolic Syndrome in Rabbit: Methodological Considerations, Development, and Assessment at JoVE.com

An Experimental Model of Diet-Induced Metabolic Syndrome in Rabbit: Methodological Considerations, Development, and Assessment

家兔膳食诱导代谢综合征的实验模型: 方法论考虑、发展和评价

我们描述的方法, 以建立一个实验模型的饮食诱导代谢综合征 (大都会) 的家兔使用高脂肪, 高蔗糖的饮食。动物发育为中央性肥胖症、轻度高血压、糖尿病前和血脂紊乱, 从而再现了人类大都会的主要成分。这种慢性模式将允许获取疾病进展的基础机制知识。

In recent years, obesity and metabolic syndrome (MetS) have become a growing problem for public health and clinical practice, given their increased ...

an-experimental-model-diet-induced-metabolic-syndrome-rabbit

Universidad San Sebastian

Research

JoVE Journal

Medicine

10.6K 次观看.  Instituto de Salud Carlos III. 我们描述的方法, 以建立一个实验模型的饮食诱导代谢综合征 (大都会) 的家兔使用高脂肪, 高蔗糖的饮食。动物发育为中央性肥胖症、轻度高血压、糖尿病前和血脂紊乱, 从而再现了人类大都会的主要成分。这种慢性模式将允许获取疾病进展的基础机制知识。

视频: An Experimental Model of Diet-Induced Metabolic Syndrome in Rabbit: Methodological Considerations, Development, and Assessment

Stem Cell Transplantation in an in vitro Simulated Ischemia/Reperfusion Model

Stem cell transplantation protocols are finding their way into clinical practice1,2,3. Getting better results, making the protocols more robust, and finding new sources for implantable cells are the focus of recent research4,5. Investigating the effectiveness of cell therapies is not an easy task and new tools are needed to investigate the mechanisms involved in the treatment process6. We designed an experimental protocol of ischemia/reperfusion in order to allow the observation of cellular connections and even subcellular mechanisms during ischemia/reperfusion injury and after stem cell transplantation and to evaluate the efficacy of cell therapy. H9c2 cardiomyoblast cells were placed onto cell culture plates7,8. Ischemia was simulated with 150 minutes in a glucose free medium with oxygen level below 0.5%. Then, normal media and oxygen levels were reintroduced to simulate reperfusion. After oxygen glucose deprivation, the damaged cells were treated with transplantation of labeled human bone marrow derived mesenchymal stem cells by adding them to the culture. Mesenchymal stem cells are preferred in clinical trials because they are easily accessible with minimal invasive surgery, easily expandable and autologous. After 24 hours of co-cultivation, cells were stained with calcein and ethidium-homodimer to differentiate between live and dead cells. This setup allowed us to investigate the intercellular connections using confocal fluorescent microscopy and to quantify the survival rate of postischemic cells by flow cytometry. Confocal microscopy showed the interactions of the two cell populations such as cell fusion and formation of intercellular nanotubes. Flow cytometry analysis revealed 3 clusters of damaged cells which can be plotted on a graph and analyzed statistically. These populations can be investigated separately and conclusions can be drawn on these data on the effectiveness of the simulated therapeutical approach.

Stem Cell Transplantation in an in vitro Simulated Ischemia/Reperfusion Model

We demonstrate how to set up an in vitro ischemia/reperfusion model and how to evaluate the effect of stem cell therapy on postischemic cardiac cells.

在干细胞移植在体外模拟缺血/再灌注模型

Stem cell transplantation protocols are finding their way into clinical practice1,2,3. Getting better results, making the protocols more robust, and ...

科研

医学

Matrix-assisted Autologous Chondrocyte Transplantation for Remodeling and Repair of Chondral Defects in a Rabbit Model

Articular cartilage defects are considered a major health problem because articular cartilage has a limited capacity for self-regeneration 1. Untreated cartilage lesions lead to ongoing pain, negatively affect the quality of life and predispose for osteoarthritis. During the last decades, several surgical techniques have been developed to treat such lesions. However, until now it was not possible to achieve a full repair in terms of covering the defect with hyaline articular cartilage or of providing satisfactory long-term recovery 2-4. Therefore, articular cartilage injuries remain a prime target for regenerative techniques such as Tissue Engineering. In contrast to other surgical techniques, which often lead to the formation of fibrous or fibrocartilaginous tissue, Tissue Engineering aims at fully restoring the complex structure and properties of the original articular cartilage by using the chondrogenic potential of transplanted cells. Recent developments opened up promising possibilities for regenerative cartilage therapies.
The first cell based approach for the treatment of full-thickness cartilage or osteochondral lesions was performed in 1994 by Lars Peterson and Mats Brittberg who pioneered clinical autologous chondrocyte implantation (ACI) 5. Today, the technique is clinically well-established for the treatment of large hyaline cartilage defects of the knee, maintaining good clinical results even 10 to 20 years after implantation 6. In recent years, the implantation of autologous chondrocytes underwent a rapid progression. The use of an artificial three-dimensional collagen-matrix on which cells are subsequently replanted became more and more popular 7-9.
MACT comprises of two surgical procedures: First, in order to collect chondrocytes, a cartilage biopsy needs to be performed from a non weight-bearing cartilage area of the knee joint. Then, chondrocytes are being extracted, purified and expanded to a sufficient cell number in vitro. Chondrocytes are then seeded onto a three-dimensional matrix and can subsequently be re-implanted. When preparing a tissue-engineered implant, proliferation rate and differentiation capacity are crucial for a successful tissue regeneration 10. The use of a three-dimensional matrix as a cell carrier is thought to support these cellular characteristics 11.
The following protocol will summarize and demonstrate a technique for the isolation of chondrocytes from cartilage biopsies, their proliferation in vitro and their seeding onto a 3D-matrix (Chondro-Gide, Geistlich Biomaterials, Wollhusen, Switzerland). Finally, the implantation of the cell-matrix-constructs into artificially created chondral defects of a rabbit's knee joint will be described. This technique can be used as an experimental setting for further experiments of cartilage repair.

An experimental technique for the treatment of chondral defects in the rabbit's knee joint is described. The implantation of autologous chondrocytes seeded on a matrix is a well-accepted method for the remodeling and repair of articular cartilage lesions providing satisfying long-term results. Matrix-assisted autologous chondrocyte transplantation (MACT) offers a standardized and clinically established implantation method.

基质辅助重塑和修复软骨缺损的自体软骨细胞移植在兔模型

Articular cartilage defects are considered a major health problem because articular cartilage has a limited capacity for self-regeneration 1. Untreated ...

Isolation and Analysis of Plasma Lipoproteins by Ultracentrifugation

Analysis of plasma lipoproteins and apolipoproteins is an essential part for the diagnosis of dyslipidemia and studies of lipid metabolism and atherosclerosis. Although there are several methods for analyzing plasma lipoproteins, ultracentrifugation is still one of the most popular and reliable methods. Because of its intact separation procedure, the lipoprotein fractions isolated by this method can be used for analysis of lipoproteins, apolipoproteins, proteomes, and functional study of lipoproteins with cultured cells in vitro. Here, we provide a detailed protocol to isolate seven lipoprotein fractions including VLDL (d&#60;1.006 g/mL), IDL (d=1.02 g/mL), LDLs (d=1.04 and 1.06 g/mL), HDLs (d=1.08, 1.10, and 1.21 g/mL) from rabbit plasma using sequential floating ultracentrifugation. In addition, we introduce the readers how to analyze apolipoproteins such as apoA-I, apoB, and apoE by SDS-PAGE and Western blotting and show representative results of lipoprotein and apolipoprotein profiles using hyperlipidemic rabbit models. This method can become a standard protocol for both clinicians and basic scientists to analyze lipoprotein functions.

Several methods have been used for analyzing plasma lipoproteins; however, ultracentrifugation is still one of the most popular and reliable methods. Here, we describe a method regarding how to isolate lipoproteins from plasma using sequential density ultracentrifugation and how to analyze the apolipoproteins for both diagnostic and research purposes.

超中心对等离子脂蛋白的分离和分析

Analysis of plasma lipoproteins and apolipoproteins is an essential part for the diagnosis of dyslipidemia and studies of lipid metabolism and ...

生物化学

The Rabbit Model of Accelerated Atherosclerosis: A Methodological Perspective of the Iliac Artery Balloon Injury

Acute coronary syndrome resulting from coronary occlusion following atherosclerotic plaque development and rupture is the leading cause of death in the industrialized world. New Zealand White (NZW) rabbits are widely used as an animal model for the study of atherosclerosis. They develop spontaneous lesions when fed with atherogenic diet; however, this requires long time of 4 - 8 months. To further enhance and accelerate atherogenesis, a combination of atherogenic diet and mechanical endothelial injury is often employed. The presented procedure for inducing atherosclerotic plaques in rabbits uses a balloon catheter to disrupt the endothelium in the left iliac artery of NZW rabbits fed with atherogenic diet. Such mechanical damage caused by the balloon catheter induces a chain of inflammatory reactions initiating neointimal lipid accumulation in a time dependent fashion. Atherosclerotic plaque following balloon injury show neointimal thickening with extensive lipid infiltration, high smooth muscle cell content and presence of macrophage derived foam cells. This technique is simple, reproducible and produces plaque of controlled length within the iliac artery. The whole procedure is completed within 20 - 30 min. The procedure is safe with low mortality and also offers high success in obtaining substantial intimal lesions. The procedure of balloon catheter induced arterial injury results in atherosclerosis within two weeks. This model can be used for investigating the disease pathology, diagnostic imaging and to evaluate new therapeutic strategies.

Animal models of atherosclerosis are essential to understand the mechanism and to investigate newer approaches to prevent plaque development or rupture, a leading cause of death in the industrialized world. This protocol uses a combination of balloon injury and cholesterol rich diet to induce atherosclerotic plaques in rabbit iliac artery.

家兔加速动脉粥样硬化模型的研究--髂动脉球囊损伤的方法学观察

Acute coronary syndrome resulting from coronary occlusion following atherosclerotic plaque development and rupture is the leading cause of death in the ...

Preclinical Model of Hind Limb Ischemia in Diabetic Rabbits

Peripheral vascular disease is a widespread clinical problem that affects millions of patients worldwide. A major consequence of peripheral vascular disease is the development of ischemia. In severe cases, patients can develop critical limb ischemia in which they experience constant pain and an increased risk of limb amputation. Current therapies for peripheral ischemia include bypass surgery or percutaneous interventions such as angioplasty with stenting or atherectomy to restore blood flow. However, these treatments often fail to the continued progression of vascular disease or restenosis or are contraindicated due to the overall poor health of the patient. A promising potential approach to treat peripheral ischemia involves the induction of therapeutic neovascularization to allow the patient to develop collateral vasculature. This newly formed network alleviates peripheral ischemia by restoring perfusion to the affected area. The most frequently employed preclinical model for peripheral ischemia utilizes the creation of hind limb ischemia in healthy rabbits through femoral artery ligation. In the past, however, there has been a strong disconnect between the success of preclinical studies and the failure of clinical trials regarding treatments for peripheral ischemia. Healthy animals typically have robust vascular regeneration in response to surgically induced ischemia, in contrast to the reduced vascularity and regeneration in patients with chronic peripheral ischemia. Here, we describe an optimized animal model for peripheral ischemia in rabbits that includes hyperlipidemia and diabetes. This model has reduced collateral formation and blood pressure recovery in comparison to a model with a higher cholesterol diet. Thus, the model may provide better correlation with human patients with compromised angiogenesis from the common co-morbidities that accompany peripheral vascular disease.

We describe a surgical procedure used to induce peripheral ischemia in rabbits with hyperlipidemia and diabetes. This surgery acts as a preclinical model for conditions experienced in peripheral artery disease in patients. Angiography is also described as a means to measure the extent of introduced ischemia and recovery of perfusion.

糖尿病家兔欣德肢体缺血的临床前模型

Peripheral vascular disease is a widespread clinical problem that affects millions of patients worldwide. A major consequence of peripheral vascular ...

Palatable Western-style Cafeteria Diet as a Reliable Method for Modeling Diet-induced Obesity in Rodents

Obesity is rapidly increasing in incidence in developed and developing countries and is known to induce or exacerbate many diseases.&#160;The health burden of obesity and its comorbid conditions highlight the need for better understanding of its pathogenesis, yet ethical constraints limit studies in humans. To this end externally valid models of obesity in laboratory animals are essential for the understanding of being overweight and obesity.&#160;While many species have been used to model the range of changes that accompany obesity in humans, rodents are most commonly used. Our laboratory has developed a western-style cafeteria diet that consistently leads to considerable weight gain and markers of metabolic disease in rodents. The diet exposes rodents to a variety of highly palatable foods to induce hyperphagia, modeling the modern western food environment. This diet rapidly induces weight gain and body fat accumulation in rats allowing for the study of effects of overeating and obesity. While the cafeteria diet may not provide the same control over macronutrient and micronutrient profile as purified high-fat or high-fat, high-sugar diets, the cafeteria diet typically induces a more severe metabolic phenotype than that observed with purified diets and is more in line with metabolic disturbances observed in the overweight and obese human population.

This protocol describes the use of a highly palatable, western-style cafeteria diet to model overeating and obesity in rodents. Here, we provide a detailed outline of food selection, preparation and measurement, and explain methodological factors that assist in generating a robust and reproducible phenotype.

美味的西式自助餐厅饮食作为模拟啮齿动物饮食诱发肥胖症的可靠方法

Obesity is rapidly increasing in incidence in developed and developing countries and is known to induce or exacerbate many diseases.&#160;The health ...

Model of Ischemia and Reperfusion Injury in Rabbits

The protocol here provides a simple, highly replicable methodology to induce in situ acute regional myocardial ischemia in the rabbit for non-survival and survival experiments. New Zealand White adult rabbit is sedated with atropine, acepromazine, butorphanol, and isoflurane. The animal is intubated and placed on mechanical ventilation. An intravenous catheter is inserted into the marginal ear vein for the infusion of medications. The animal is pre-medicated with heparin, lidocaine, and lactated Ringer&#39;s solution. A carotid cut-down is performed to obtain arterial line access for blood pressure monitoring. Select physiologic and mechanical parameters are monitored and recorded by continuous real-time analysis.
With the animal sedated and fully anesthetized, either a fourth intercostal space small left thoracotomy (survival) or midline sternotomy (non-survival) is performed. The pericardium is opened, and the left anterior descending (LAD) artery is located.
A polypropylene suture is passed around the second or third diagonal branch of the LAD artery, and the polypropylene filament is threaded through a small vinyl tube, forming a snare. The animal is subjected to 30 min of regional ischemia, achieved by occluding the LAD by tightening the snare. Myocardial ischemia is confirmed visually by regional cyanosis of the epicardium. Following regional ischemia, the ligature is loosened, and the heart is allowed to re-perfuse.
For both survival and non-survival experiments, the myocardial function can be assessed via an echocardiography (ECHO) measurement of the fractional shortening. For non-survival studies, data from sonomicrometry collected using three digital piezoelectric ultrasonic probes implanted within the ischemic area and the left ventricle developed pressure (LVDP) using an apically inserted left ventricle (LV) catheter can be continuously acquired for evaluating the regional and global myocardial function, respectively.
For survival studies, the incision is closed, a left needle thoracentesis is performed for pleural air evacuation, and postoperative pain control is achieved.

The present study demonstrates a highly reproducible animal model of acute regional myocardial ischemia and reperfusion injury in rabbits using a left mini-thoracotomy for survival cases or a midline sternotomy for non-survival cases.

兔子缺血和再灌注损伤模型

The protocol here provides a simple, highly replicable methodology to induce in situ acute regional myocardial ischemia in the rabbit for non-survival and ...

<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 ...

家兔膳食诱导代谢综合征的实验模型: 方法论考虑、发展和评价

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在干细胞移植在体外模拟缺血/再灌注模型

基质辅助重塑和修复软骨缺损的自体软骨细胞移植在兔模型

超中心对等离子脂蛋白的分离和分析

家兔加速动脉粥样硬化模型的研究--髂动脉球囊损伤的方法学观察

糖尿病家兔欣德肢体缺血的临床前模型

美味的西式自助餐厅饮食作为模拟啮齿动物饮食诱发肥胖症的可靠方法

兔子缺血和再灌注损伤模型

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

小鼠糖尿病心肌病模型的建模与评价