hyperinsulinemic – euglycemic दबाना इंसुलिन कार्रवाई का आकलन करने के लिए "सोने के मानक 'है. इंसुलिन एक निरंतर ग्लूकोज तेज उत्तेजक दर पर संचार होता है. इस ड्रॉप काउंटर करने के लिए संचार exogenous ग्लूकोज की मात्रा इंसुलिन संवेदनशीलता का संकेत है. यहाँ प्रक्रिया एक सचेत, अनर्गल चूहे पर किया जाता है.
Type 2 diabetes (T2D) is rapidly rising in prevalence. Characterized by either inadequate insulin production or the inability to utilize insulin produced, T2D results in elevated blood glucose levels. The “gold-standard” in assessing insulin sensitivity is a hyperinsulinemic-euglycemic clamp or insulin clamp. In this procedure, insulin is infused at a constant rate resulting in a drop in blood glucose. To maintain blood glucose at a constant level, exogenous glucose (D50) is infused into the venous circulation. The amount of glucose infused to maintain homeostasis is indicative of insulin sensitivity. Here, we show the basic clamp procedure in the chronically catheterized, unrestrained, conscious rat. This model allows blood to be collected with minimal stress to the animal. Following the induction of anesthesia, a midline incision is made and the left common carotid artery and right jugular vein are catheterized. Inserted catheters are flushed with heparinized saline, then exteriorized and secured. Animals are allowed to recover for 4-5 days prior to experiments, with weight gain monitored daily. Only those animals who regain weight to pre-surgery levels are used for experiments. On the day of the experiment, rats are fasted and connected to pumps containing insulin and D50. Baseline glucose is assessed from the arterial line and used a benchmark throughout the experiment (euglycemia). Following this, insulin is infused at a constant rate into the venous circulation. To match the drop in blood glucose, D50 is infused. If the rate of D50 infusion is greater than the rate of uptake, a rise in glucose will occur. Similarly, if the rate is insufficient to match whole body glucose uptake, a drop will occur. Titration of glucose continues until stable glucose readings are achieved. Glucose levels and glucose infusion rates during this stable period are recorded and reported. Results provide an index of whole body insulin sensitivity. The technique can be refined to meet specific experimental requirements. It is further enhanced by the use of radioactive tracers that can determine tissue specific insulin-stimulated glucose uptake as well as whole body glucose turnover.
शुरू में मानव में इंसुलिन संवेदनशीलता की जांच के लिए विकसित की, दबाना प्रक्रिया अब प्रयोगशाला चूहों और चूहों सहित अन्य प्रजातियों के लिए अनुकूलित किया गया. इंसुलिन प्रतिरोध की जांच पशु मॉडल में इंसुल…
The authors have nothing to disclose.
इस अध्ययन स्वास्थ्य अनुसंधान और जीनोम Canada.JS कनाडा के संस्थानों द्वारा समर्थित किया गया है वेतन अलबर्टा विरासत Foundationfor मेडिकल रिसर्च, हार्ट और स्ट्रोक फाउंडेशन कनाडा डायबिटीज एसोसिएशन Canadaand के से समर्थन पुरस्कार रखती है. डॉ. डेविड Wasserman और घसियारा प्रयोगशाला के लिए इस प्रक्रिया को पढ़ाने के लिए Bingle Bracy के लिए विशेष धन्यवाद.
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 |