Name: assessing whole-body lipid-handling capacity in mice
Uploaded: 2020-11-24T14:00:00
Description: Watch this Scientific Journal Video about Assessing Whole-Body Lipid-Handling Capacity in Mice at JoVE.com

这项工作得到国家卫生研究院（NIH）的支持，授予R00-DK114498，美国农业部（美国农业部）向Y.Z提供3092-51000-062。

根据贝勒医学院（BCM）机构动物护理和使用委员会批准的动物护理和实验协议，动物被安置在标准化条件下。动物被喂食标准或特殊的饮食，水广告脂质，并保持12小时的日夜循环。
1. 测量禁食血清脂
<ol><li>下午5点以后将老鼠转移到一个新的笼子里，并快速免费取水，过夜（实验前大约16个小时的禁食）。通宵禁食确保完全排空小鼠的胃肠道。 注意：老鼠在禁食时吃粪便...

<ol>
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这三个分析描述了实验室中强有力的功能，并考虑了几个关键问题。夜间禁食是确定禁食血清脂质水平和口腔脂内耐受性测试所必需的。对于口腔脂内耐受性测试，在室温下旋转血液以尽量减少脂肪层的形成至关重要，尤其是在 1 小时和 2 小时的时间点：重要的是不要丢弃这个脂肪层，如果它形成。确保将超高肽与脂质层一起转移，并轻轻地将它们混合在一起，以进行甘油三酯测定。
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碳水化合物和脂质是能量代谢的两大基质。异常脂质代谢导致许多人类疾病，包括II型糖尿病、心血管疾病、脂肪肝疾病和癌症。膳食脂质，主要是甘油三酯，通过肠道吸收到淋巴系统，并进入心脏1附近的血小龙静脉循环。脂质由血液中的脂蛋白颗粒携带，脂肪酸脂肪通过脂蛋白脂酶在肌肉和脂肪组织2等外周器官的作用而释放。剩余的富含胆固醇的残留颗粒由肝脏3清除。老鼠在实验室中被广泛用作研究脂质代谢的研究模型。拥有全面的遗传工具集和相对较短的繁殖周期，它们是研究脂质如何被吸收、合成和代谢的有力模型。
由于脂质代谢的复杂性，复杂的脂质学研究或同位素示踪研究通常用于量化脂质物种或脂质相关代谢通量和命运的集合4，5。这给没有专门设备或专业知识的研究人员带来了巨大的挑战。在本文中，我们提出了三个测试，在使用具有技术挑战性的技术之前，这些测试可以作为初始测试。它们是小鼠的非终端程序，因此非常有助于识别脂质处理能力的潜在差异和缩小受影响的过程。
首先，测量禁食血清脂质分子可以帮助确定小鼠的整体脂质特征。老鼠应该禁食，因为许多脂质物种在饭后会上升，而且增加的程度受到饮食成分的强烈影响。许多脂质分子，包括总胆固醇、甘油三酯和非酯化脂肪酸（NEFA），都可以使用商业套件和可读取吸血的板读器进行测量。
其次，口服脂质耐受性测试将脂质处理能力评估为吸收和代谢的净效应。口服的脂内导致循环甘油三酯水平激增（1至2小时），之后血清甘油三酯水平恢复到基底水平（4至6小时）。此检测提供了有关鼠标处理外源性脂质的程度的信息。心脏、肝脏和棕色脂肪组织是甘油三酯的活跃消费者，而白色脂肪组织将其储存为能量储备。这些功能的变化将导致测试结果的差异。
最后，促进脂溶解以调动储存的脂质被认为是减肥的可能策略。脂肪组织中的β3-肾神经受体信号通路在脂肪细胞脂解中起着重要作用，人类遗传学已经识别出与肥胖相关的3-肾上腺受体中功能丧失的多态性Trp64Arg。CL 316，243，一种特异且有效的β-肾上腺素受体激动剂，刺激脂肪组织脂解和甘油释放。评估鼠标对CL 316，243的反应可以提供关于化合物的开发、改进和理解的宝贵信息。
总的来说，这些测试可以用作小鼠脂质代谢状态变化的初始屏幕。它们是为仪器和试剂的可访问性而选择的。通过这些检测结果，研究人员可以形成动物代谢适应的整体图景，并决定更复杂和更有针对性的方法。

<table>
	<tbody>
		<tr>
			<td>20% Intralipid</td>
			<td>Sigma Aldrich</td>
			<td>I141</td>
			<td></td>
		</tr>
		<tr>
			<td>BD Slip Tip Sterile Syringes 1ml</td>
			<td>Shaotong</td>
			<td>B07F1KRMYN</td>
			<td></td>
		</tr>
		<tr>
			<td>CL 316,243 Hydrate</td>
			<td>Sigma-Aldrich</td>
			<td>C5976</td>
			<td></td>
		</tr>
		<tr>
			<td>Curved Feeding Needles (18 Gauge)</td>
			<td>Kent Scientific</td>
			<td>FNC-18-2-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Free Glycerol Reagent</td>
			<td>Sigma Aldrich</td>
			<td>F6428</td>
			<td></td>
		</tr>
		<tr>
			<td>Glycerol Standard Solution</td>
			<td>Sigma</td>
			<td>G7793</td>
			<td></td>
		</tr>
		<tr>
			<td>HR SERIES NEFA-HR(2)COLOR REAGENT A</td>
			<td>Fujifilm Wako Diagnostics</td>
			<td>999-34691</td>
			<td></td>
		</tr>
		<tr>
			<td>HR SERIES NEFA-HR(2)COLOR REAGENT B</td>
			<td>Fujifilm Wako Diagnostics</td>
			<td>991-34891</td>
			<td></td>
		</tr>
		<tr>
			<td>HR SERIES NEFA-HR(2)SOLVENT A</td>
			<td>Fujifilm Wako Diagnostics</td>
			<td>995-34791</td>
			<td></td>
		</tr>
		<tr>
			<td>HR SERIES NEFA-HR(2)SOLVENT B</td>
			<td>Fujifilm Wako Diagnostics</td>
			<td>993-35191</td>
			<td></td>
		</tr>
		<tr>
			<td>Ketamine</td>
			<td>Vedco</td>
			<td>50989-161-06</td>
			<td></td>
		</tr>
		<tr>
			<td>Matrix Plus Chemistry Reference Kit</td>
			<td>Verichem</td>
			<td>9500</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro Centrifuge Tubes</td>
			<td>Fisher Scientific</td>
			<td>14-222-168</td>
			<td></td>
		</tr>
		<tr>
			<td>Microhematrocrit Capillary Tube, Not Heparanized</td>
			<td>Fisher Scientific</td>
			<td>22-362-574</td>
			<td></td>
		</tr>
		<tr>
			<td>NEFA STANDARD SOLUTION</td>
			<td>Fujifilm Wako Diagnostics</td>
			<td>276-76491</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate Buffered Saline</td>
			<td>Boston Bioproducts</td>
			<td>BM-220</td>
			<td></td>
		</tr>
		<tr>
			<td>Thermo Scientific Triglycerides Reagent</td>
			<td>Fisher Scientific</td>
			<td>TR22421</td>
			<td></td>
		</tr>
		<tr>
			<td>Total Cholesterol Reagents</td>
			<td>Thermo Scientifi</td>
			<td>TR13421</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylazine</td>
			<td>Henry Schein</td>
			<td>11695-4022-1</td>
			<td></td>
		</tr>
	</tbody>
</table>

assessing whole-body lipid-handling capacity in mice

评估脂质代谢是评估代谢功能的基石，它被认为是活体代谢研究的关键。脂质是许多不同的分子的一类，其合成和代谢涉及许多通路。需要一个起点来评估营养和肥胖研究的脂质血脂。本文描述了三种简单易行的方法，这些方法几乎不需要专业知识或实践来掌握，并且大多数实验室都可以将其改编为筛查小鼠的脂质代谢异常。这些方法是（1）测量几个禁食血清脂质分子使用商业套件（2）通过口服脂质耐受性测试，以评估饮食脂质处理能力，（3）评估对药物化合物CL 316，243的反应，在小鼠。这些方法将共同提供小鼠脂质处理能力的高水平概述。

我们用三段摘录显示，每次检测都提供了有关小鼠脂质代谢的宝贵信息。对于C57BL/6J雄性小鼠来说，从8周大开始接受8周高脂肪饮食（HFD）喂养的挑战，总胆固醇水平显著升高，而血清甘油三酯和NEFA则没有（表1），这表明血液中的甘油三酯和NEFA不是主要受膳食脂肪挑战的调节。在第二组小鼠中，C57BL/6J和C57BL6/N子系统从8周大开始喂养HFD8周。他们的血清甘油三酯水平在口腔脂内挑战后?...

Watch this Scientific Journal Video about Assessing Whole-Body Lipid-Handling Capacity in Mice at JoVE.com

Assessing Whole-Body Lipid-Handling Capacity in Mice

本文为评估小鼠脂质代谢提供了三种简单易行的检测。

评估小鼠全身脂质处理能力

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Research

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Programmed Electrical Stimulation in Mice

程控电刺激小鼠

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Transthoracic Echocardiography in Mice

经胸超声心动图在小鼠

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在斑马鱼模型的胎儿酒精暴露评估致畸的变化

Fetal alcohol syndrome (FAS) is a severe manifestation of embryonic exposure to ethanol. It presents with characteristic defects to the face and organs, ...

Protocol for Long Duration Whole Body Hyperthermia in Mice

持续时间较长的小鼠全身热疗协议

Hyperthermia is a general term used to define the increase in core body temperature above normal. It is often used to describe the increased core body ...

Assessing Changes in Volatile General Anesthetic Sensitivity of Mice after Local or Systemic Pharmacological Intervention

局部或全身药物干预后评估的变化小鼠中挥发性全身麻醉药敏感性

One desirable endpoint of general anesthesia is  the state of unconsciousness, also known as hypnosis. Defining the hypnotic state in animals is  less ...

A Method to Study the Impact of Chemically-induced Ovarian Failure on Exercise Capacity and Cardiac Adaptation in Mice

一个方法研究的化学诱导卵巢早衰的运动能力和适应心脏的影响在小鼠

The risk of cardiovascular disease (CVD) increases in post-menopausal women, yet, the role of exercise, as a preventative measure for CVD risk in ...

Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice

颈动脉输液的紫杉烷类化合物在小鼠体内的药代动力学和药效学分析

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study ...

Assessing Specificity of Anticancer Drugs In Vitro

Assessing Specificity of Anticancer Drugs In Vitro

评估抗癌药物的特异性<em&gt;体外</em

A procedure for assessing specificity of anticancer drugs in vitro using cultures containing both tumor and non-tumor cells is demonstrated. The key ...

Phosphorus-31 Magnetic Resonance Spectroscopy: A Tool for Measuring In Vivo Mitochondrial Oxidative Phosphorylation Capacity in Human Skeletal Muscle

Phosphorus-31 Magnetic Resonance Spectroscopy: A Tool for Measuring In Vivo Mitochondrial Oxidative Phosphorylation Capacity in Human Skeletal Muscle

磷31磁共振波谱分析：一种测量工具<em&gt;在体内</em&gt;线粒体氧化磷酸化能力人骨骼肌

Skeletal muscle mitochondrial oxidative phosphorylation (OXPHOS) capacity, which is critically important in health and disease, can be measured in vivo ...

Whole Body and Regional Quantification of Active Human Brown Adipose Tissue Using 18F-FDG PET/CT

Whole Body and Regional Quantification of Active Human Brown Adipose Tissue Using 18F-FDG PET/CT

18F-fdg Petct 对活性人棕色脂肪组织的全身和区域定量

In endothermic animals, brown adipose tissue (BAT) is activated to produce heat for defending body temperature in response to cold. BAT&#39;s ability to ...