Name: hyperinsulinemic-euglycemic clamp in the conscious rat
Uploaded: 2011-02-07T17:00:00
Description: Watch this Scientific Journal Video about Hyperinsulinemic-Euglycemic Clamp in the Conscious Rat at JoVE.com

这项研究是由加拿大健康研究和基因组Canada.JS研究院的支持下，拥有来自阿尔伯塔省文物Foundationfor医学研究，心脏及中风基金会，加拿大糖尿病协会的Canadaand的薪金支持奖。特别感谢博士大卫沃瑟曼和宾格尔布雷西此过程希勒实验室教学。

答：手术导管，动脉和静脉循环第1部分：动脉和静脉导管的制备<ol><li>切15厘米PE - 50（内部直径0.58毫米（0.023“）x外径0.965毫米（0.038”）。硅胶管切1毫米节（0.76毫米（0.030“）内部直径x1.65毫米（0.065“）外径），使用限制珠。限制珠防止拉出导管，一次到位的老鼠。 </li><li>微解剖钳插入管腔的限制珠的技巧和镊子轻轻托起的秘诀除了伸展扩大开放。使用另一微解剖钳，滑?...

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	<li>Halseth, A. E., Bracy, D. P., Wasserman, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Limitations+to+basal+and+insulin-stimulated+skeletal+muscle+glucose+uptake+in+the+high-fat-fed+rat.">Limitations to basal and insulin-stimulated skeletal muscle glucose uptake in the high-fat-fed rat.</a> Am J Physiol Endocrinol Metab. 279, E1064-E1071 (2000).</li><li>Halseth, A. E., Bracy, D. P., Wasserman, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Limitations+to+muscle+glucose+uptake+due+to+high+fat+feeding.">Limitations to muscle glucose uptake due to high fat feeding.</a> Am. J. Physiol. 279, E1064-E1071 (2000).</li><li>Muniyappa, R., Lee, S., Chen, H., Quon, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+approaches+for+assessing+insulin+sensitivity+and+resistance+in+vivo:+advantages,+limitations,+and+appropriate+usage.">Current approaches for assessing insulin sensitivity and resistance in vivo: advantages, limitations, and appropriate usage.</a> Am J Physiol Endocrinol Metab. 294, 15-26 (2008).</li><li>Cho, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insulin+resistance+and+a+diabetes+mellitus-like+syndrome+in+mice+lacking+the+protein+kinase+Akt2+(PKB+beta).">Insulin resistance and a diabetes mellitus-like syndrome in mice lacking the protein kinase Akt2 (PKB beta).</a> Science. 292, 1728-1731 (2001).</li><li>Dubois, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+SHP-1+protein+tyrosine+phosphatase+negatively+modulates+glucose+homeostasis.">The SHP-1 protein tyrosine phosphatase negatively modulates glucose homeostasis.</a> Nat Med. 12, 549-556 (2006).</li><li>Pacini, G., Thomaseth, K., Ahren, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contribution+to+glucose+tolerance+of+insulin-independent+vs.+insulin-dependent+mechanisms+in+mice.">Contribution to glucose tolerance of insulin-independent vs. insulin-dependent mechanisms in mice.</a> Am J Physiol Endocrinol Metab. 281, 693-703 (2001).</li><li>Herbach, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dominant-negative+effects+of+a+novel+mutated+Ins2+allele+causes+early-onset+diabetes+and+severe+beta-cell+loss+in+Munich+Ins2C95S+mutant+mice.">Dominant-negative effects of a novel mutated Ins2 allele causes early-onset diabetes and severe beta-cell loss in Munich Ins2C95S mutant mice.</a> Diabetes. 56, 1268-1276 (2007).</li><li>Maeda, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diet-induced+insulin+resistance+in+mice+lacking+adiponectin/ACRP30.">Diet-induced insulin resistance in mice lacking adiponectin/ACRP30.</a> Nat Med. 8, 731-737 (2002).</li><li>Potenza, M. A., Marasciulo, F. L., Tarquinio, M., Quon, M. J., Montagnani, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treatment+of+spontaneously+hypertensive+rats+with+rosiglitazone+and/or+enalapril+restores+balance+between+vasodilator+and+vasoconstrictor+actions+of+insulin+with+simultaneous+improvement+in+hypertension+and+insulin+resistance.">Treatment of spontaneously hypertensive rats with rosiglitazone and/or enalapril restores balance between vasodilator and vasoconstrictor actions of insulin with simultaneous improvement in hypertension and insulin resistance.</a> Diabetes. 55, 3594-3603 (2006).</li><li>Ayala, J. E., Bracy, D. P., McGuinness, O. P., Wasserman, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Considerations+in+the+design+of+hyperinsulinemic-euglycemic+clamps+in+the+conscious+mouse.">Considerations in the design of hyperinsulinemic-euglycemic clamps in the conscious mouse.</a> Diabetes. 55, 390-397 (2006).</li><li>Wasserman, D. H., Ayala, J. E., McGuinness, O. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lost+in+translation.">Lost in translation.</a> Diabetes. 58, 1947-1950 (2009).</li></ol>

最初调查在人类胰岛素敏感性的发展，现在已经被钳制程序适应其他物种，包括实验大鼠和小鼠。调查胰岛素抵抗的动物模型提供了一个了解对胰岛素的敏感性和相关的病症的病理生理，以及确定的临床应用价值1,2的治疗性干预的大量援助。已受聘的几种方法，以评估在动物体内胰岛素的敏感性。这些技术包括糖耐量试验（GTTs）的变化和胰岛素糖耐量试验（ITTS）4-6以及?...

<table border="1"><tbody><tr valign="top"><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论（可选） </td></tr><tr valign="top"><td> Intramedic聚乙烯管（PE - 50） </td><td> Fisher Scientific则</td><td> 14 - 170 - 12B </td><td>内部直径0.58毫米（0.023“）的X外径0.965毫米（0.038”） </td></tr><tr valign="top"><td>道康宁的硅橡胶实验室套管</td><td> Fisher Scientific则</td><td> 11 - 189 - 15C </td><td>内部直径0.76毫米（0.030“）的X1.65毫米外径（0.065”） </td></tr><tr valign="top"><td>聚乙烯小号- 50 - HL医用导管</td><td>哈佛器械</td><td> PY2 72-1251 </td><td>内部直径为3.2mm（0.125“）的X4.7毫米外径（0.1875”） </td></tr><tr valign="top"><td>乐泰超级胶水</td><td>大和玩具</td><td> 32237 </td><td>凝胶控制</td></tr><tr valign="top"><td>无菌手术刀片</td><td> VWR </td><td> BD371610 </td><td></td></tr><tr valign="top"><td>弧形微解剖钳</td><td>乔治Tiemann＆CO。 </td><td> 160-20 </td><td> × 2 </td></tr><tr valign="top"><td>直微解剖钳</td><td>乔治Tiemann＆CO。 </td><td> 160-15 </td><td> × 2 </td></tr><tr valign="top"><td>弯止血钳</td><td>乔治Tiemann＆CO。 </td><td> 105-1135 </td><td> × 2 </td></tr><tr valign="top"><td>直止血钳</td><td>乔治Tiemann＆CO。 </td><td> 105-1130 </td><td> × 2 </td></tr><tr valign="top"><td>止血提示警卫</td><td>罗宾斯仪器公司</td><td> 15.09-2-004 </td><td></td></tr><tr valign="top"><td>直外科剪刀</td><td>乔治Tiemann＆CO。 </td><td> 105-402 </td><td></td></tr><tr valign="top"><td> VENOJECT多样品露儿适配器</td><td>泰尔茂医疗产品</td><td> 810127A </td><td> 21计，1英寸</td></tr><tr valign="top"><td>无菌导管导引</td><td>碧迪</td><td> 406999 </td><td></td></tr><tr valign="top"><td> 14号钝针</td><td>碧迪</td><td> 511310 </td><td> 14号，2英寸</td></tr><tr valign="top"><td>无菌手术缝合</td><td>强生公司医药产品</td><td> 1679H </td><td>丝绸，大小3-0 </td></tr><tr valign="top"><td>非无菌手术缝合线</td><td> Angiotech制药公司</td><td> SP116 </td><td>丝绸，大小4-0 </td></tr><tr valign="top"><td>棉签</td><td> VWR </td><td> 10806-005 </td><td></td></tr><tr valign="top"><td> 4ply纱布垫</td><td> VWR </td><td> CA43845 - 062 </td><td></td></tr><tr valign="top"><td>小动物无绳快船</td><td>哈佛器械</td><td> 729063 </td><td></td></tr><tr valign="top"><td>异氟醚</td><td>卤烃产品公司</td><td> IPN - 45 </td><td></td></tr><tr valign="top"><td>麻醉车</td><td>森医疗等行业，公司</td><td></td><td></td></tr><tr valign="top"><td> 70％的乙醇</td><td> Fisher Scientific则</td><td> HC - 1000 </td><td></td></tr><tr valign="top"><td>优碘消毒液</td><td>西药配送中心有限公司</td><td> 105267 </td><td></td></tr><tr valign="top"><td> 11型加注射泵</td><td>哈佛器械</td><td> 702208 </td><td></td></tr><tr valign="top"><td>不锈钢油管耦合</td><td>哈佛器械</td><td> 72-4434 </td><td> 23计，0.3英寸</td></tr><tr valign="top"><td>不锈钢管插头</td><td>哈佛器械</td><td> 72-4436 </td><td> 23计，0.5英寸</td></tr><tr valign="top"><td>不锈钢钝性针</td><td> Instech实验室，公司</td><td> LS22 </td><td> 22号</td></tr><tr valign="top"><td> 60度Y型连接器</td><td>小零件</td><td> STCY - 22 - 05 </td><td> 22号</td></tr><tr valign="top"><td> CritSpin微血细胞比容离心机</td><td>虹膜来样加工</td><td> CS12 </td><td></td></tr><tr valign="top"><td>迷你离心机</td><td> Fisher Scientific则</td><td> 05-090-100 </td><td></td></tr><tr valign="top"><td>微型离心管</td><td> VWR </td><td> 53550-778 </td><td></td></tr><tr valign="top"><td> 50毫升聚丙烯离心管</td><td> VWR</td><td> 89004-364 </td><td></td></tr><tr valign="top"><td> 1毫升塑料滑提示注射器</td><td>碧迪</td><td> 309602 </td><td></td></tr><tr valign="top"><td> 3毫升塑料Luerlok提示注射器</td><td>碧迪</td><td> 309585 </td><td></td></tr><tr valign="top"><td>肝素抗凝注射液</td><td>西药配送中心有限公司</td><td> 102824 </td><td>生产商：LEO制药公司浓。 1000国际单位</td></tr><tr valign="top"><td> EDTA溶液</td><td> Promega公司的公司</td><td> V4231 </td><td> 0.5米，pH值8.0 </td></tr><tr valign="top"><td>盐水</td><td>西药配送中心有限公司</td><td> ABB7983154 </td><td>制造商：Hospira公司0.9％的氯化钠</td></tr><tr valign="top"><td> 50％葡萄糖</td><td> Vetoquinol </td><td> 8DEX012D </td><td></td></tr><tr valign="top"><td>优泌林 - R </td><td>礼来公司</td><td> HI - 210 </td><td> 100U/ml </td></tr><tr valign="top"><td> 1毫升胰岛素注射器</td><td>碧迪</td><td> 309311 </td><td></td></tr><tr valign="top"><td> Fisherbrand *造血密封胶</td><td> Fisher Scientific则</td><td> 02-678 </td><td></td></tr><tr valign="top"><td> Fisherbrand * Microhematocrit毛细管</td><td> Fisher Scientific则</td><td> 22-362-574 </td><td></td></tr><tr valign="top"><td>单键超试纸</td><td> LIFESCAN，公司</td><td> AW 085 - 314H </td><td></td></tr><tr valign="top"><td>单键Ultra血糖仪</td><td> LIFESCAN，公司</td><td> AW 085 - 314B </td><td></td></tr><tr valign="top"><td>钠Pentobarbitol </td><td>塞瓦三特，石兽</td><td> 1715 138 </td><td>浓。 54.7mg/ml </td></tr><tr valign="top"><td>红实验室标签带</td><td> VWR </td><td> 89097-932 </td><td></td></tr><tr valign="top"><td>蓝实验室标签带</td><td> VWR </td><td> 89097-936 </td><td></td></tr><tr valign="top"><td>电子秤</td><td> Fisher Scientific则</td><td> 01-913-88 </td><td></td></tr><tr valign="top"><td>涡旋</td><td> VWR </td><td> 58815-234 </td><td></td></tr><tr valign="top"><td>定时器</td><td> VWR </td><td> 62344-641 </td><td></td></tr></tbody></table>

hyperinsulinemic-euglycemic clamp in the conscious rat

2型糖尿病（T2D）患病率迅速上升。其特点是无论是生产胰岛素不足或无法利用血糖水平升高的胰岛素产生，T2D结果。 “黄金标准”在评估胰岛素敏感性，是高胰岛素 - 正常血糖钳或胰岛素钳夹。在此过程中，胰岛素注入在一个恒定的，导致血糖下降率。为了保持血糖在一个恒定的水平，外源性葡萄糖（D50）注入静脉循环。注入的葡萄糖量保持动态平衡胰岛素敏感性的指标。在这里，我们展示在长期插管，奔放，有意识地鼠的基本钳程序。这种模式使血液收集，以最小的压力动物。麻醉诱导后，正中切口和左颈总动脉和右颈静脉插管。插入导管用肝素生理盐水冲洗，然后exteriorized和安全。动物被允许恢复实验前4-5天，每天监测体重增加。只有那些谁复胖到术前水平的动物用于实验。在一天的实验，老鼠被禁食含有胰岛素和D50泵相连。基线葡萄糖是从动脉行评估整个实验（血糖正常），并用一个基准。在此之后，是在一个恒定的速率进入静脉循环注入胰岛素。为了配合在血糖下降，D50的注入。如果D50的输液速度大于吸收率，血糖的上升会发生。同样的，如果利率是不够的，以配合整个身体对葡萄糖的摄取，下降会发生。葡萄糖的滴定继续进行，直到达到稳定血糖读数。血糖水平，在这个稳定的时期葡萄糖输注速率记录和报告。结果提供了一个全身胰岛素敏感性指数。该技术可以改进，以满足特定的实验要求。它进一步增强了使用的放射性示踪剂，可以决定组织特异性胰岛素刺激的葡萄糖摄取以及全身葡萄糖营业额。

Watch this Scientific Journal Video about Hyperinsulinemic-Euglycemic Clamp in the Conscious Rat at JoVE.com

Hyperinsulinemic-Euglycemic Clamp in the Conscious Rat

高胰岛素 - 正常血糖钳是胰岛素的作用进行评估的“金标准”。胰岛素是注入在一个恒定的刺激葡萄糖摄取率。注入外源性葡萄糖的量，以应对这一下降是胰岛素敏感性的指标。这里的程序是一种自觉的，奔放的大鼠上进行。

在自觉大鼠高胰岛素 - 正常血糖钳

Type 2 diabetes (T2D) is rapidly rising in prevalence. Characterized by either inadequate insulin production or the inability to utilize insulin produced, ...

Research

JoVE Journal

Medicine

Hyperinsulinemic-euglycemic Clamps in Conscious, Unrestrained Mice

在自觉的，奔放的小鼠正常血糖高胰岛素 - 夹钳

Type 2 diabetes is characterized by a defect in insulin action. The hyperinsulinemic-euglycemic clamp, or insulin clamp, is widely considered the "gold ...

科研

医学

Angiogenesis in the Ischemic Rat Lung

在缺血大鼠肺的血管生成

The  adult lung is perfused by both the systemic bronchial artery and the entire  venous return flowing through the pulmonary arteries. In most lung ...

Slow-release Drug Delivery through Elvax 40W to the Rat Retina: Implications for the Treatment of Chronic Conditions

通过40W的Elvax缓释给药的大鼠视网膜：对慢性疾病的治疗

Diseases of the retina are difficult to treat as the retina lies deep within the eye. Invasive methods of drug delivery are often needed to treat these ...

Catheterization of the Carotid Artery and Jugular Vein to Perform Hemodynamic Measures, Infusions and Blood Sampling in a Conscious Rat Model

颈动脉和颈静脉导管在有意识大鼠模型进行血流动力学措施，输液和血液采样

The success of a small animal model to study critical illness is, in part, dependent on the ability of the model to simulate the human condition. ...

Transcutaneous Assessment of Renal Function in Conscious Rodents

在鼠害自觉肾功能的评估皮

Glomerular filtration rate (GFR) is the gold standard to assess overall kidney function. However, traditional methods to evaluate GFR are cumbersome and ...

The Dimethylnitrosamine Induced Liver Fibrosis Model in the Rat

该二甲基亚硝胺诱发肝纤维化模型大鼠

Four to six week old, male Wistar rats were used to produce animal models of liver fibrosis. The process requires four weeks of administration of 10 mg/kg ...

Method of Direct Segmental Intra-hepatic Delivery Using a Rat Liver Hilar Clamp Model

直接节段性肝内交付使用大鼠肝门部夹具模型方法

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ...

Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy

Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy

静脉大鼠部分肝移植的体内血管定向70% 肝切除术

Split liver transplantation and living liver donor liver transplantation were developed in the clinic to utilize liver organs in a more efficient manner. ...

Assessment of Respiratory Function in Conscious Mice by Double-chamber Plethysmography

双室 Plethysmography 对清醒小鼠呼吸功能的评价

Air volume changes created by a conscious subject breathing spontaneously within a body box are at the basis of plethysmography, a technique used to ...

Repetitive Blood Sampling from the Subclavian Vein of Conscious Rat

Rats are widely used in pharmacokinetics (PK) and toxicokinetic (TK) studies that need to collect a certain amount of blood at specific time points to ...