在自觉大鼠高胰岛素 - 正常血糖钳
Summary
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。
Abstract
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。
Protocol
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。 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?…
Discussion
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?…
Disclosures
The authors have nothing to disclose.
Acknowledgements
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。
Materials
| Name of the reagent | Company | Catalogue number | Comments (optional) |
| Intramedic Polyethylene Tubing (PE-50) | Fisher Scientific | 14-170-12B | Internal diameter of .58mm (.023″) x Outer diameter of .965mm (.038″) |
| Dow Corning Silastic Laboratory Tubing | Fisher Scientific | 11-189-15C | Internal diameter of .76mm (0.030″) x Outer diameter of 1.65mm (0.065″) |
| Tygon S-50-HL Medical Tubing | Harvard Apparatus | PY2 72-1251 | Internal diameter of 3.2mm (0.125″) x Outer diameter of 4.7mm (0.1875″) |
| Loctite Super Glue | Grand & Toy | 32237 | Gel Control |
| Sterile Surgical Blade | VWR | BD371610 | |
| Curved Micro Dissecting Forceps | George Tiemann & Co. | 160-20 | x 2 |
| Straight Micro Dissecting Forceps | George Tiemann & Co. | 160-15 | x 2 |
| Curved Hemostat | George Tiemann & Co. | 105-1135 | x 2 |
| Straight Hemostat | George Tiemann & Co. | 105-1130 | x 2 |
| Hemostat Tip Guards | Robbins Instruments, Inc. | 15.09-2-004 | |
| Straight Surgery Scissors | George Tiemann & Co. | 105-402 | |
| VENOJECT Multi-Sample Luer Adapter | Terumo Medical Products | 810127A | 21 guage, 1 in. |
| Sterile Catheter Introducer | Becton Dickinson | 406999 | |
| 14-gauge Blunt Needle | Becton Dickinson | 511310 | 14 guage, 2 in. |
| Sterile Surgical Suture | Johnson & Johnson Medical Products | 1679H | Silk, size 3-0 |
| Non-Sterile Surgical Suture | Angiotech Pharmaceuticals, Inc. | SP116 | Silk, size 4-0 |
| Cotton Swabs | VWR | 10806-005 | |
| 4ply Gauze Pads | VWR | CA43845-062 | |
| Small Animal Cordless Clippers | Harvard Apparatus | 729063 | |
| Isoflurane | Halocarbon Products Corp. | IPN-45 | |
| Anesthetic Cart | Benson Medical Industries, Inc. | ||
| 70 % Ethanol | Fisher Scientific | HC-1000 | |
| Betadine Antiseptic Solution | Western Drug Distribution Centre Ltd. | 105267 | |
| Model 11 Plus Syringe Pump | Harvard Apparatus | 702208 | |
| Stainless Steel Tubing Couplers | Harvard Apparatus | 72-4434 | 23 gauge, 0.3 in. |
| Stainless Steel Tubing Plugs | Harvard Apparatus | 72-4436 | 23 gauge, 0.5 in. |
| Stainless Steel Blunt Needles | Instech Laboratories, Inc. | LS22 | 22 gauge |
| 60 Degree Y-Connectors | Small Parts | STCY-22-05 | 22 gauge |
| CritSpin Micro-hematocrit Centrifuge | Iris Sample Processing | CS12 | |
| Mini Centrifuge | Fisher Scientific | 05-090-100 | |
| Micro Centrifuge Tubes | VWR | 53550-778 | |
| 50ml polypropylene centrifuge tubes | VWR | 89004-364 | |
| 1ml Plastic Slip Tip Syringes | Becton Dickinson | 309602 | |
| 3ml Plastic Luerlok Tip Syringes | Becton Dickinson | 309585 | |
| Heparin Anticoagulant Injection | Western Drug Distribution Centre Ltd. | 102824 | Manufacturer: LEO Pharma Inc. Conc. 1000 IU |
| EDTA Solution | Promega Corp. | V4231 | 0.5 M, pH 8.0 |
| Saline | Western Drug Distribution Centre Ltd. | ABB7983154 | Manufacturer: Hospira 0.9% Sodium Chloride |
| 50% Dextrose | Vetoquinol | 8DEX012D | |
| Humulin-R | Eli Lilly | HI-210 | 100U/ml |
| 1ml Insulin Syringes | Becton Dickinson | 309311 | |
| Fisherbrand* Hemato-Seal Sealant | Fisher Scientific | 02-678 | |
| Fisherbrand* Microhematocrit Capillary Tubes | Fisher Scientific | 22-362-574 | |
| One Touch Ultra Test Strips | LifeScan, Inc. | AW 085-314H | |
| One Touch Ultra Blood Glucose Meter | LifeScan, Inc. | AW 085-314B | |
| Sodium Pentobarbitol | Ceva Sante Animale | 1715 138 | Conc. 54.7mg/ml |
| Red Laboratory Labeling Tape | VWR | 89097-932 | |
| Blue Laboratory Labeling Tape | VWR | 89097-936 | |
| Weigh Scale | Fisher Scientific | 01-913-88 | |
| Vortex | VWR | 58815-234 | |
| Timer | VWR | 62344-641 |
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Tags
Hyperinsulinemic-euglycemic ClampType 2 DiabetesInsulin SensitivityBlood Glucose LevelsInsulin ProductionInsulin UtilizationGold-standardHyperinsulinemic-euglycemic Clamp ProcedureInfusion Of InsulinDrop In Blood GlucoseExogenous GlucoseVenous CirculationHomeostasisInsulin Sensitivity AssessmentChronically Catheterized RatAnesthesia InductionCarotid Artery CatheterizationJugular Vein CatheterizationHeparinized Saline FlushWeight Gain MonitoringFastingInsulin And D50 Infusion
Cite This Article
Hughey, C. C., Hittel, D. S., Johnsen, V. L., Shearer, J. Hyperinsulinemic-Euglycemic Clamp in the Conscious Rat. J. Vis. Exp. (48), e2432, doi:10.3791/2432 (2011).