Name: study of in vivo glucose metabolism in high-fat diet-fed mice using oral glucose tolerance test (ogtt) and insulin tolerance test (itt)
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这项研究得到了维也纳市市长的医疗科学基金和Österreichische 德国 Laboratoriumsmedizin 和 Klinische 瓦的支持。

这里描述的所有方法都已得到维也纳医科大学动物护理和使用委员会的批准, 并根据欧洲实验动物科学协会联合会 (FELASA) 进行。请注意, 本议定书所述的所有程序都应在机构和政府批准之后以及在技术上精通的工作人员执行。
1. 风喂养小鼠
注意: 在12小时的光/暗循环中保持所有 C57BL/6J 小鼠, 可以免费获得食物和水。
<ol><li>在6周的年龄, 将老鼠放在风 (40-6...

<ol>
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随着糖尿病和相关疾病在世界人口中的高患病率, 对疾病的分子机制、预防和治疗的研究有很强的要求19。该协议描述了建立风小鼠的成熟方法, 一种用于代谢研究的健壮动物模型, 以及 OGTT 和 ITT 的传导, 它们是评估全身代谢改变的有力工具。如胰岛素抵抗。本文所提出的方法可能有助于研究疑似基因、环境因素以及药理、膳食、物理或基因治疗对全身葡萄糖代谢的作用<sup class="x...

在发达世界, 由于缺乏体力活动和加工食品的过量消费, 肥胖和糖尿病已经达到了流行病的程度, 这是由于迅速的城市化、工业化和全球化所造成的影响。虽然研究胰岛素抵抗和它的疾病, 如高脂血症和动脉粥样硬化, 已经在过去的几十年中获得突出, 复杂的生物机制, 调节代谢在健康和疾病仍然不完全了解, 仍然迫切需要新的治疗方法来预防和治疗这些疾病1。
胰岛素, 它的 counter-regulatory 激素胰高血糖素作为主要调节细胞能量供应和营养素平衡, 从而也保持适当的系统性血糖浓度2。葡萄糖本身是胰腺β细胞分泌胰岛素的主要刺激之一, 而其他营养素、体液因子以及神经输入则进一步改变了这种反应。胰岛素通过促进过剩的血糖扩散到肌肉和脂肪细胞, 并进一步激活糖酵解以及蛋白质或脂肪酸的合成, 从而触发了联邦政府的合成代谢过程。此外, 胰岛素抑制糖, 抑制肝脏葡萄糖的输出。慢性过量能量消耗和 meta-inflammation 导致高和周围胰岛素抵抗由于胰岛素受体表达的下调以及下游信号通路的改变, 从而导致受损对胰岛素介导的葡萄糖处理的敏感性以及对肝脏葡萄糖分泌不足的抑制作用3,4,5,6。
广泛的动物模型与遗传, 营养, 或实验性诱导疾病已被证明是优秀的工具, 研究胰岛素抵抗的分子机制和各种形式的糖尿病以及其伴随的疾病7.一个典型的例子是广泛使用, 并建立了良好的风诱导小鼠模型, 这是特点是快速体重增加, 由于饮食摄入与降低代谢效率相结合, 导致胰岛素抵抗8,9. 在动物模型和人类中, 空腹血糖和胰岛素水平的升高, 以及对葡萄糖管理的耐受性受损, 常被用于胰岛素抵抗和其他葡萄糖系统改变的指标代谢.因此, 监测血糖和胰岛素水平的基础状态或刺激后, 容易接近读数。
本议定书概述了风喂养小鼠的产生, 以及两种常用的方法, 口服葡萄糖耐受试验 (OGTT) 和胰岛素抵抗试验 (ITT), 这是有用的表征代谢表型和调查葡萄糖代谢的改变。我们详细地描述了 OGTT, 它评估了口服葡萄糖负荷和胰岛素分泌的处理时间。此外, 我们还提供了关于如何通过监测血糖浓度来对胰岛素的反应来检查全身胰岛素的行为的指导。本文中描述的协议已被广泛采用, 并已在多个研究中使用10、11、12。除了轻微的修改, 这可能有助于增加成功, 我们提供了实验设计和数据分析的指导方针, 以及有用的提示, 以避免潜在的陷阱。本文所描述的协议可以是非常强大的工具, 以研究遗传, 药理, 饮食, 和其他环境因素对全身葡萄糖代谢的影响, 并对其相关的疾病, 如胰岛素抵抗。除了刺激葡萄糖或胰岛素, 其他各种化合物可用于刺激, 根据个人研究的目的。虽然在这份手稿的范围之外, 许多其他下游应用可以在抽取的血样上进行, 例如对血糖和胰岛素以外的血液值进行分析 (例如、血脂和脂蛋白图谱) 以及详细代谢标志物的分析 (例如, 通过定量实时聚合酶链式反应 (PCR), 免疫印迹分析, 酶联免疫吸附试验 (ELISA))。进一步的流式细胞仪和荧光活化细胞分类 (组) 可用于研究不同单细胞种群的影响, 同时也可用于不分析。
总的来说, 我们提供了一个简单的协议来生成一个风诱导的小鼠模型, 同时进一步描述了研究全身代谢变化的两个强有力的方法, OGTT 和 ITT, 这可以成为有用的工具, 研究疾病的发病机制和开发新的治疗方法, 特别是在代谢相关疾病领域, 如胰岛素抵抗 & 糖尿病。

<table>
	<tbody>
		<tr>
			<td>Mouse strain: C57BL/6J</td>
			<td>The Jackson Laboratory</td>
			<td>664</td>
			<td>LFD/HFD</td>
		</tr>
		<tr>
			<td>Accu Chek Performa - Glucometer</td>
			<td>Roche</td>
			<td>6870228</td>
			<td>OGTT/ITT</td>
		</tr>
		<tr>
			<td>Accu Chek Performa - Strips</td>
			<td>Roche</td>
			<td>6454038</td>
			<td>OGTT/ITT</td>
		</tr>
		<tr>
			<td>D-(+)-Glucose solution</td>
			<td>Sigma-Aldrich</td>
			<td>G8769</td>
			<td>OGTT</td>
		</tr>
		<tr>
			<td>Actrapid - Insulin</td>
			<td>Novo Nordisk</td>
			<td>417642</td>
			<td>ITT</td>
		</tr>
		<tr>
			<td>Reusable Feeding Needles</td>
			<td>Fine Science Tools</td>
			<td>#18061-22</td>
			<td>OGTT; 22 gauge (-24 gauge for young mice)</td>
		</tr>
		<tr>
			<td>Omnifix-Fine dosing syringes</td>
			<td>Braun</td>
			<td>9161406V</td>
			<td>OGTT/ITT</td>
		</tr>
		<tr>
			<td>Sterican Insulin needle (30G x 1/3&#34;; &#248; 0.30 x 13 mm)</td>
			<td>Braun</td>
			<td>304000</td>
			<td>ITT; lean mice</td>
		</tr>
		<tr>
			<td>Sterican (G 27 x 3/4&#34;; &#248; 0.40 x 20 mm)&#160;&#160;</td>
			<td>Braun</td>
			<td>4657705</td>
			<td>ITT; mice on HFD</td>
		</tr>
		<tr>
			<td>96 Well PCR Plates, non-skirted, flexible</td>
			<td>Braintree Scientific, Inc.</td>
			<td>SP0016</td>
			<td>OGTT</td>
		</tr>
		<tr>
			<td>Ultrasensitive Mouse Insulin ELISA kit</td>
			<td>Crystam Chem</td>
			<td>90080</td>
			<td>OGTT</td>
		</tr>
		<tr>
			<td>Rodent Diet with 60% kcal% fat</td>
			<td>Research Diets Inc</td>
			<td>D12492</td>
			<td>mice on HFD</td>
		</tr>
		<tr>
			<td>Rodent Diet with 10% kcal% fat.</td>
			<td>Research Diets Inc</td>
			<td>D12450B</td>
			<td>mice on LFD</td>
		</tr>
		<tr>
			<td>BRAND micro haematocrit capillary</td>
			<td>Sigma-Aldrich</td>
			<td>BR749321</td>
			<td>OGTT/ITT</td>
		</tr>
		<tr>
			<td>Vaseline - creme</td>
			<td>Riviera</td>
			<td>P1768677</td>
			<td>OGTT/ITT</td>
		</tr>
	</tbody>
</table>

study of in vivo glucose metabolism in high-fat diet-fed mice using oral glucose tolerance test (ogtt) and insulin tolerance test (itt)

肥胖是2型糖尿病发病的最重要的单一危险因素, 这种疾病的特点是抗胰岛素刺激的葡萄糖摄取和全身糖代谢的严重失代偿。尽管在了解葡萄糖代谢方面取得了相当大的进展, 但它在健康和疾病方面的调控的分子机制仍然 under-investigated, 而新的预防和治疗糖尿病的方法是迫切需要的。饮食衍生葡萄糖刺激胰腺分泌胰岛素, 这是主要调节器的细胞合成过程中的美联储状态, 从而平衡血糖水平保持系统的能量状态。慢性过度触发 meta-inflammation, 导致周围胰岛素受体相关信号的改变, 从而降低对胰岛素介导的葡萄糖处理的敏感性。这些事件最终导致空腹血糖和胰岛素水平升高以及葡萄糖耐受减少, 这反过来又成为胰岛素抵抗的重要指标。在这里, 我们提出了一个协议的产生和代谢特征的高脂饮食 (风) 喂养小鼠作为一个常用的模型, 饮食诱导胰岛素抵抗。我们详细说明了口服葡萄糖耐受试验 (OGTT), 它监测的周围处理的口服葡萄糖负荷和胰岛素分泌随着时间的推移。此外, 我们提出了一个协议的胰岛素耐受性测试 (ITT), 以监测全身胰岛素行为。这些方法和它们的下游应用代表了强有力的工具来表征小鼠的一般代谢表型, 以及专门评估葡萄糖代谢的变化。它们可能在胰岛素抵抗、糖尿病和肥胖的广泛研究领域中特别有用, 以便更好地了解发病机制, 并测试治疗干预的效果。

图 1说明了在饮食上对小鼠进行代谢分的示意图时间表。在大约6周的年龄, 老鼠应该放在一个风, 而 LFD 组可以作为对照组。重要的是, 体重应该每周确定, 以观察是否有预期的体重增加。任何形式的压力 (例如、噪音或攻击性的男性行为) 都会影响体重增加, 应立即消除。每组小鼠的饮食实验应包括至少10小鼠, 因为这些饮食-实验是耗时的, 和异?...

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Study of In Vivo Glucose Metabolism in High-fat Diet-fed Mice Using Oral Glucose Tolerance Test OGTT and Insulin Tolerance Test ITT

口服葡萄糖耐受试验 (OGTT) 和胰岛素耐受试验 (ITT) 对高脂饮食喂养小鼠体内葡萄糖代谢的影响研究 (英文)

本文描述了高脂喂养小鼠的生成和代谢特性, 作为饮食诱导的胰岛素抵抗和肥胖的模型。它还有详细的协议来执行口服葡萄糖耐受试验和胰岛素耐受试验, 监测葡萄糖代谢的全身变化在体内。

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Study of In Vivo Glucose Metabolism in High-fat Diet-fed Mice Using Oral Glucose Tolerance Test (OGTT) and Insulin Tolerance Test (ITT)

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