Name: assessment of the metabolic effects of isocaloric 2:1 intermittent fasting in mice
Uploaded: 2019-11-27T13:30:00
Description: Watch this Scientific Journal Video about Assessment of the Metabolic Effects of Isocaloric 2:1 Intermittent Fasting in Mice at JoVE.com

K.-H.K得到了加拿大心脏和中风基金会（G-18-0022213）、J.P.Bickell基金会和渥太华大学心脏研究所启动基金的支持;H.-K.S.得到了加拿大卫生研究院（PJT-162083）、鲁本和海伦·丹尼斯学者以及太阳生活金融新研究员的资助，该奖项来自班廷和最佳糖尿病中心（BBDC）和自然科学和加拿大工程研究理事会（RGPIN-2016-06610）。R.Y.K.得到了渥太华大学心脏病研究捐赠基金的一个研究金的支持。J.H.L.得到了NSERC博士奖学金和安大略省研究生奖学金的支持。Y.O.获得UOHI颁发的研究生奖和伊丽莎白二世女王科技研究生奖学金的支持。

这里的所有方法和协议都经渥太华大学动物护理和兽医服务（ACVS）动物护理委员会和动物基因组学中心（TCP）批准，并符合加拿大动物护理理事会的标准。应当指出，此处描述的所有程序都应在机构和政府批准下以及由技术熟练的工作人员执行。所有小鼠都被安置在温度和湿度控制室的标准通风笼中，具有 12 小时/12 小时的光/暗循环（21~22 °C，正常住房湿度为 30%-60%），并可自由取水。雄性C57BL/6...

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有充分记载，IF对人类和动物的各种疾病8、15、16、17、18、19等多种疾病都具有有益的健康作用。其潜在的机制，如自噬和肠道微生物群，最近已被阐明。提出的方案描述了小鼠的等分2：1 IF方案，用于研究IF对饮食引起的肥胖和相关代谢功能障碍的?...

现代生活方式与更长的每日食物摄入时间和较短的禁食期1相关。这促成了当前的全球肥胖流行，代谢的劣势在人类身上出现。禁食在整个人类历史中一直实践，其多样化的健康益处包括延长寿命，减少氧化损伤，和优化能量平衡2，3。在几种练习禁食的方法中，定期能量剥夺，称为间歇性禁食（IF），是一种流行的饮食方法，由于其简单易行的疗养，被一般人广泛采用。最近对临床前和临床模型的研究表明，IF可以提供与长期禁食和热量限制相当的健康益处，这表明IF可以作为肥胖和代谢疾病2、3、4、5的潜在治疗策略。
IF方案在禁食持续时间和频率方面有所不同。隔天禁食（即1天喂食/1天禁食;1：1 IF）是啮齿动物最常用的IF疗法，以研究其对肥胖、心血管疾病、神经退行性疾病等有益的健康影响。然而，如先前研究6、7所示，在我们的能量摄入分析8，1：1IF导致喂养不足（+80%）中进一步确认由于缺乏足够的喂食时间来弥补能量损失。这使得不清楚1：1 IF所赋予的健康益处是通过限制卡路里还是改变饮食模式来调节的。因此，一个新的IF方案已经开发出来，并在这里显示，包括2天喂食/1天禁食（2：1 IF）模式，这为小鼠提供了足够的时间来补偿食物摄入量（+99%）和体重。然后，将这些小鼠与活动（AL）组进行比较。这种方案能够在野生型小鼠没有热量降低的情况下检查等热IF的影响。
相反，在显示反馈行为改变的小鼠模型中，AL 喂食可能不是比较和检查 2：1 IF 效果的适当控制条件。例如，由于ob/ob小鼠（一种常用的肥胖基因模型）由于缺乏瘦素调节食欲和饱口福而表现出吞咽困难，因此与使用 AL 喂养的ob/ob小鼠相比，2：1 IF 的小鼠表现出热量摄入减少 20%。因此，为了正确检查和比较在ob/ob小鼠中 IF 的影响，需要采用配对喂养组作为适当的对照组。
总体而言，提供了一个全面的协议来执行等分2：1 IF，包括使用对馈送控制。进一步证明，等热2：1 IF保护小鼠免受高脂肪饮食引起的肥胖和/或代谢功能障碍在野生型和ob/ob小鼠。该协议可用于检查 2：1 IF 对各种病理状况（包括神经系统疾病、心血管疾病和癌症）的有益健康影响。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Comprehensive Lab Animal Monitoring System (CLAMS)</td>
			<td>Columbus Instruments</td>
			<td></td>
			<td>Indirect calorimeter</td>
		</tr>
		<tr>
			<td>D-(+)-Glucose solution</td>
			<td>Sigma-Aldrich</td>
			<td>G8769</td>
			<td>For GTT</td>
		</tr>
		<tr>
			<td>EchoMRI 3-in-1</td>
			<td>EchoMRI</td>
			<td>EchoMRI 3-in-1</td>
			<td>Body composition analysis</td>
		</tr>
		<tr>
			<td>Glucometer and strips</td>
			<td>Bayer</td>
			<td>Contour NEXT</td>
			<td>These are for GTT and ITT experiments</td>
		</tr>
		<tr>
			<td>High Fat Diet (45% Kcal% fat)</td>
			<td>Research Diets Inc.</td>
			<td>#D12451</td>
			<td>3.3 Kcal/g</td>
		</tr>
		<tr>
			<td>High Fat Diet (60% Kcal% fat)</td>
			<td>Research Diets Inc.</td>
			<td>#D12452</td>
			<td>4.73 Kcal/g</td>
		</tr>
		<tr>
			<td>Insulin</td>
			<td>El Lilly</td>
			<td>Humulin R</td>
			<td>For ITT</td>
		</tr>
		<tr>
			<td>Mouse Strain: B6.Cg-Lepob/J</td>
			<td>The Jackson Laboratory</td>
			<td>#000632</td>
			<td>Ob/Ob mouse</td>
		</tr>
		<tr>
			<td>Mouse Strain: C57BL/6J</td>
			<td>The Jackson Laboratory</td>
			<td>#000664</td>
			<td></td>
		</tr>
		<tr>
			<td>Normal chow (17% Kcal% fat)</td>
			<td>Harlan</td>
			<td>#2918</td>
			<td></td>
		</tr>
		<tr>
			<td>Scale</td>
			<td>Mettler Toledo</td>
			<td></td>
			<td>Body weight and food intake measurement</td>
		</tr>
	</tbody>
</table>

assessment of the metabolic effects of isocaloric 2:1 intermittent fasting in mice

间歇性禁食（IF），一种涉及周期性能量限制的饮食干预，被认为具有许多益处和抵消代谢异常。到目前为止，已经记录了不同类型的国际直供模型，其禁食和喂食期不同。然而，解释结果具有挑战性，因为许多模型都涉及时间和卡路里限制策略的多因素贡献。例如，通常用作啮齿动物 IF 方案的隔天禁食模式可能导致喂养不足，这表明这种干预对健康的益处可能通过热量限制和禁食-再喂养周期进行调节。最近，已经成功地证明，2：1 IF，包括1天的禁食，然后2天的喂养，可以提供防止饮食引起的肥胖和代谢改善，而不减少总热量摄入。这里介绍的是这种等量2：1IF干预小鼠的协议。还描述了一个双喂食（PF）协议，需要检查小鼠模型与改变的饮食行为，如高吞咽。使用 2：1 IF 方案，证明等量IF可降低体重增加、改善葡萄糖平衡和增加能量消耗。因此，这种疗法可能有助于研究IF对各种疾病状况的健康影响。

图1显示了24小时禁食后的进餐分析以及1：1和2：1间间歇禁食的比较。24小时禁食期导致体重减少±10%，在重新喂食2天后完全恢复（图1A）。24小时禁食期在随后2天的重新喂食期间诱发吞咽过度（图1B）。然而，将1：1替代日禁食和2：1间间歇禁食之间的能量摄入量进行比较后发现，1：1 IF的重新喂食期的1天?...

Watch this Scientific Journal Video about Assessment of the Metabolic Effects of Isocaloric 2:1 Intermittent Fasting in Mice at JoVE.com

Assessment of the Metabolic Effects of Isocaloric 2:1 Intermittent Fasting in Mice

本文描述了异热2：1间歇性禁食的详细方案，以保护和治疗野生型和ob/ob小鼠的肥胖和葡萄糖代谢受损。

小鼠间间间禁食的代谢效应评估 2：1 间歇禁食

Intermittent fasting (IF), a dietary intervention involving periodic energy restriction, has been considered to provide numerous benefits and counteract ...

两到一个间歇性禁食方案包括一天的禁食，然后两天的喂养，使得研究饮食干预的影响涉及定期能量限制，而不是卡路里摄入的影响。与交替的一天禁食不同，在喂养下两到一次间歇性禁食为小鼠提供了足够的时间来充分补偿禁食过程中的能量损失和体重减轻。这使得科学家能够检查定期禁食的影响，与卡路里摄入量无关。两到一个间歇性禁食促进许多好处，在肥胖和相关代谢功能障碍，包括受损的葡萄糖平衡和脂肪肝疾病的饮食。该协议可以很容易地调整为五到两个饮食方案，包括五天的喂养和两天的禁食或限时喂养，其中获得食物的时间是每天八小时。如果使用小鼠模型，显示替代喂养行为，如超噬菌体，则副食性喂养可能不是适当的控制条件。在这种情况下，需要配对喂养规则来检查和比较间歇性禁食的影响。首先准备一个正常的饮食或高脂肪饮食的精益和饮食诱发肥胖小鼠模型。分别使用刻度和 EchomRI 测量每七周大雄性 C57BL/6J 小鼠的基线体重和组成。根据体重和身体组成结果随机和平均地将小鼠分成两组：ad libitum 或 AL、间歇性禁食或 IF 组。然后，每个笼子放置两到三只老鼠，确保老鼠可以自由获得饮用水。在开始 IF 方案之前，允许小鼠在一周内适应新的笼环境。当准备开始禁食期时，在12：00将老鼠转移到一个干净的笼子里，在12：00.m。向 AL 组提供加权量的食物。没有食物给 IF 组。24小时后，称重两组小鼠以及AL笼中剩余的食物。下午 12：00 .m，向 AL 和 IF 组提供加权量的食物。一旦喂养期过期，测量小鼠的重量和剩余食物。在研究期间重复循环。在实验期间设置一对喂养或 PF 控制组，以确保卡路里与 IF 的独立比较。测量 IF 组消耗的食物量。将它均匀地分成三部分，每天向 PF 控件提供一部分。当与饮食行为改变的动物（如高噬细胞）合作时，每天向对照组提供同等数量的食物，以防止老鼠同时食用食物至关重要。要分析小鼠的身体组成，打开身体组成分析仪，让机器打开两三个小时进行预热。在使用它之前，运行系统测试以确保测量的准确性。如有必要，使用菜籽油校准系统。测量每只鼠标的体重。然后，把它放在小动物圆柱架上。插入分隔符以在测量过程中限制鼠标的物理移动。将支架放入车身成分分析仪中。运行扫描程序，这可能需要 90 到 120 秒。然后，从设备中取出支架。将鼠标放回笼子。进行葡萄糖和胰岛素耐受性测试，检查间歇性禁食对葡萄糖平衡的影响。通宵或六小时禁食后，对每只小鼠进行葡萄糖或胰岛素的内向注射。然后，使用血糖计测量注射后 0、5、15、30、60 和 120 分钟的血糖。为了研究间歇性禁食对能量代谢的影响，在一个周期的 IF 内进行间接热量测量。在老鼠适应系统后，通过从漏斗和笼子底部取出食物和面包屑来快速清除它们。在两天的再喂养期24小时后重新引入食物。在 IF 周期结束时，将小鼠带回原始笼子，并收集程序中的数据。禁食24小时的小鼠体重减轻了10%，经过两天的再喂养后完全恢复。在比较一到一和二到一间间歇性禁食之间的能量摄入时，发现一天的再进气期不足以弥补禁食时热量损失。与AL相比，IF治疗导致野生型小鼠体重下降，以正常周或高脂肪饮食喂养，食物摄入量无显著差异。身体组成分析表明，如果减少脂肪质量而不改变野生型小鼠的瘦质量。对食对控制实验证实，IF减少体重增加并不是由于野生型小鼠能量摄入的改变。然而，在基因肥胖，ob/ob，小鼠对照组是无法区分的，IF组在体重和身体组成。葡萄糖和胰岛素耐受性测试表明，高脂肪饮食如果小鼠表现出显著改善的葡萄糖平衡相比，高脂肪饮食AL和高脂肪饮食对食小鼠。在 ob/ob 小鼠中，与具有未受影响的胰岛素敏感性的对照组相比，IF 组表现出显著改善的葡萄糖处理与较小的葡萄糖偏移。IF在野生型小鼠中注意到的另一个代谢作用是高氧消耗，表明能量消耗，特别是在喂养期间。小鼠的喂养行为可能受到环境因素的影响，包括房屋密度。因此，在研究过程中，保持每个笼子的相同数量的小鼠数量是重要的。使用新的鼠标模型时，建议在执行间歇性禁食实验之前检查进给行为。异体二对一间歇性禁食可轻松应用于其他疾病模型，包括糖尿病、心脏病、动脉硬化和神经系统疾病。

Research

JoVE Journal

Biology

Stable Isotopic Profiling of Intermediary Metabolic Flux in Developing and Adult Stage Caenorhabditis elegans

Stable Isotopic Profiling of Intermediary Metabolic Flux in Developing and Adult Stage Caenorhabditis elegans

在开发和成年阶段的中介代谢通量的稳定同位素分析线虫</em

Stable isotopic profiling has long permitted sensitive investigations of the metabolic consequences of genetic mutations and/or pharmacologic therapies in ...

科研

生物学

Assessment of Calcium Sparks in Intact Skeletal Muscle Fibers

钙火花的评估在正常肌肉纤维

Maintaining homeostatic Ca2+ signaling is a fundamental physiological process in living cells. Ca2+ sparks are the elementary units of Ca2+ signaling in ...

Measurement of Metabolic Rate in Drosophila using Respirometry

Measurement of Metabolic Rate in Drosophila using Respirometry

代谢率的测量<em&gt;果蝇</em&gt;使用呼吸运动

Metabolic disorders are a frequent problem affecting human health. Therefore, understanding the mechanisms that regulate metabolism is a crucial ...

Method for the Assessment of Effects of a Range of Wavelengths and Intensities of Red/near-infrared Light Therapy on Oxidative Stress In Vitro

Method for the Assessment of Effects of a Range of Wavelengths and Intensities of Red/near-infrared Light Therapy on Oxidative Stress In Vitro

对于波长和红/近红外光治疗对氧化应激强度的范围影响的评估方法<em&gt;在体外</em

Red/near-infrared light therapy (R/NIR-LT), delivered by laser or light emitting diode (LED), improves functional and morphological outcomes in a range of ...

Quantitation of Endothelial Cell Adhesiveness In Vitro

Quantitation of Endothelial Cell Adhesiveness In Vitro

内皮细胞粘附性定量<em&gt;在体外</em

One of the cardinal processes of inflammation is the infiltration of immune cells from the lumen of the blood vessel to the surrounding tissue. This ...

Isolation of Mitochondria from Minimal Quantities of Mouse Skeletal Muscle for High Throughput Microplate Respiratory Measurements

从小鼠骨骼肌高通量微孔板呼吸测量最低数量的隔离线粒体

Dysfunctional skeletal muscle mitochondria play a role in altered metabolism observed with aging, obesity and Type II diabetes. Mitochondrial ...

Gap Junctional Intercellular Communication: A Functional Biomarker to Assess Adverse Effects of Toxicants and Toxins, and Health Benefits of Natural Products

间隙连接通讯：天然产物的生物标志物的功能，以评估毒物和毒素的不利影响，以及健康的益处

This protocol describes a scalpel loading-fluorescent dye transfer (SL-DT) technique that measures intercellular communication through gap junction ...

Mixed Primary Cultures of Murine Small Intestine Intended for the Study of Gut Hormone Secretion and Live Cell Imaging of Enteroendocrine Cells

小鼠小肠的混合原代培养拟肠激素分泌的研究和活细胞成像的肠内分泌细胞的供

The gut is the largest endocrine organ of the body, with hormone-secreting enteroendocrine cells located along the length of the gastrointestinal ...

A Method to Assess Bacteriocin Effects on the Gut Microbiota of Mice

评估细菌素对小鼠肠道微生物的影响的方法

Very intriguing questions arise with our advancing knowledge on gut microbiota composition and the relationship with health, particularly relating to the ...

Medium-throughput Screening Assays for Assessment of Effects on Ca2+-Signaling and Acrosome Reaction in Human Sperm

Medium-throughput Screening Assays for Assessment of Effects on Ca2+-Signaling and Acrosome Reaction in Human Sperm

中通量筛选法评估对精子中 ca2 +信号和反应的影响

Ca2+-signaling is essential to normal sperm cell function and male fertility. Similarly, the acrosome reaction is vital for the ability of a human sperm ...