笔者想承认DHMRI CLAS照顾动物的工作人员的大力支持下，基金经理丹尼尔·佩拉尔塔先生和主治兽医，Glicerio伊格纳西奥博士，DVM MRCVS。

所有动物程序由位于北卡罗莱纳州研究园（NCRC）戴维H Murdock的研究所IACUC批准，根据协议14-017，糖尿病和糖尿病前期/胰岛素抵抗和治疗的功效的非人灵长类动物模型的表征，以改善胰岛素敏感性和代谢功能。 1.动物的选择和学习做准备<ol><li>基于每月的食物摄入量和体重记录选择饮食和体重稳定的动物。 注:食欲都展出了近期的下跌动物不应该被定性，直到他们的食物摄入量已经稳定。 <ol><li>对于成熟的动物（&gt; 5 - 6年），不要选择动物的尸体连续两个月之间的权重超过10％不等，而不首先检查了猴子和排除不是改变为重戏剧性的变化代谢状态等原因。 </li></ol></li><li>对于血糖，胰岛素和C-pepti德分析，准备与蛋白酶抑制剂（抑肽酶和DPP4I）K 2 EDTA样品收集管。 注:DPP4I +抑肽酶鸡尾酒提供了抑制蛋白酶的广谱这应该是必需的额外的分析物（胰高血糖素，GLP-1）。在该附加的分析物没有收集的情况下，血管应该以同样的方式，以保持样品收集在一致性来制备。对于不验证该方法测定样品应根据制造商的建议分别收集。 <ol><li> 100毫克冻干抑肽酶用10ml DPP4I混合准备蛋白酶抑制剂。加10μl抑肽酶+ DPP4I混合物到每个管血的采集血液每毫升。 </li><li>额外的10微升蛋白酶抑制剂混合物添加到各管中可能集合超龄。在-20°C，直到使用存储处理的血管。保持湿冰管的procedu期间重新并在4℃下旋转。 </li><li>使用血清分离管中，在基线标准血清化学分析，收集血液样品。对于全血细胞计数（CBC），使用标准的K 2 EDTA无蛋白酶抑制剂。使用离心管到血样之后等分试样的血浆和血清已降速。 </li><li>与动物识别，日期，程序，时间点和样品体积正确标记血管和冷冻管。与在血浆中的分析物为合适的测定标记冷冻管。 </li></ol></li><li>通过注入0.15毫升每毫升肝素1000 USP单位到生理盐水250毫升袋子制备肝素化盐水冲洗。得到0.06毫克肝素/ ml的溶液。绘制40 - 60毫升该溶液放入盐水锁样品之间冲洗。绘制额外1毫升和5ml成单独的注射器的输注之前和之后冲洗葡萄糖输注端口，分别。 </li></ol> 2。动物镇静和准备<ol><li>从动物的笼子里取出的食物在手术前不低于14小时，不超过18小时。 注:重要的是，动物被禁食的程序，以避免在血糖值的任何餐后变化。这也是一个防范措施，避免反流胃内容物的吸入麻醉时。 </li><li>稳重动物的IVGTT过程用氯胺酮肌肉注射作为全身麻醉剂的持续时间，在10毫克/公斤。根据需要，在操作过程中30分钟，或 - 以20间隔 - （10毫克/千克5）管理附加氯胺酮。 <ol><li>称量镇静动物。将动物在加热过程表横向躺着的位置。 </li><li>监测临床参数，每15至20分钟，以确保动物在麻醉的稳定面。测量心脏速率（100 - 200 BPM）和SPO 2（&gt; 92％）与脉搏血氧仪。测量呼吸频率（20 - 50次/ min）W，<html>第i个秒表，计数呼吸视觉或用手超过十五秒钟，乘以4。测量温度（&gt; 97°F）直肠。显示器周围的牙龈和嘴唇（潮湿，粉红色）粘膜的颜色。 </li></ol></li><li>准备两个插管部位。用推剪修剪从其中导管将被插入的感兴趣的区域的毛发，并用氯己定的交替磨砂和70％的酒精消毒的整个区域。 <ol><li>将一个导管中的左侧或右侧头部或隐静脉的区域中，并将其与一个三路活塞附加到肝素盐水冲洗（0.06毫克肝素/毫升）。这是在采样抽血站点。 </li><li>放置第二个导管中另一个腿或臂在头侧或隐静脉的区域中，并附加一个端口。使用本网站的葡萄糖输液。使用小1毫升冲洗肝素生理盐水到插管专利保持葡萄糖输液前。 </li></ol></li></ol>e_title"&gt; 3。IVGTT过程注:IVGTT过程包括8抽血取样时间点（ 见表1）。 <ol><li>取基线样品，并使用手持血糖测计仪来测量空腹血糖水平。为了评估动物的总体健康状况获取标准化学分析为CBC血清样品，以及全血样品。收集血浆样品以测定使用验证与猕猴按照制造商的说明8,9使用的试剂盒从基线样品血糖和胰岛素水平。 注:以具有从套管采取任何样本的采血以除去残留的血液或肝素在插管的死空间之前采取为0.5ml预拉伸是很重要的。 </li><li>获得基线样品后，注入50％葡萄糖（250毫克/千克）的剂量在30秒到葡萄糖输注端口。 注意:高剂量模型（500毫克/千克），可以使用虽然剂量应以使纵向比较固定跨程序。 <ol><li>冲洗用5ml肝素生理盐水的输液口，以确保不存在葡萄糖留在端口。输液的到底是T0。有技术人员更换手套，如从输注残余葡萄糖可能污染后续血样。 </li></ol></li><li>第一次输注后的采样时间点是在T3分钟，从葡萄糖输注结束，接着T5分钟，T7分钟，T10分钟，T15分钟，T20分钟，和最后一个样本的时间点是在T30分钟。从每个时间点与基线样品测定血糖和胰岛素水平收集血浆（参见步骤3.1）。 <ol><li>在T3分钟的时间点，使用手持血糖测计仪再次检查血糖水平。 注意:在基线的血糖仪读数和T3只确认葡萄糖输液。在T3分钟的时间点的血糖值应为〜100毫克/分升高共mpared到空腹基线血糖水平。 </li></ol></li></ol> 4.动物恢复和来样加工<ol><li>取出插管和压力施加到T30分钟时间点后，插入导管位点的止血。监测动物，直到它已恢复了神志，坐起来。发售饲料一旦动物完全恢复。 </li><li>立即将每个全血样品放入冰K 2 EDTA管。离心机以3000rpm在4℃收集的10分钟内的温度。分装血浆样品到冷冻管，冷冻并储存在-80℃直至分析。 <ol><li>允许血液进入标准血清化学分析的血清管在室温下静置不少于20分钟，并在在室温下3,000rpm下离心前不超过30分钟。冻结血清样本采集之前的48小时内检测。 注:冷藏收集CBC分析直至全血样本收集24小时内赛义德。 </li></ol></li></ol> 5.数据处理<ol><li>建立血浆胰岛素和葡萄糖的曲线之后，确定从自然对数高于基线16，17的血糖值的斜率的葡萄糖清除率ķ。 注:健康NHP可以预期有一个葡萄糖清除率ķ远高于1，通常比2以上时，作为健康的动物将常在30分钟内恢复到其基线血糖值。由于胰岛素的产生脱落，葡萄糖清除率k，便会下降更为显着，跌破1。 </li><li>计算的AUC作为一个整体，作为代表的时间点之间的每个线段的曲线下面积的梯形的面积的总和的总和，通过T30的16，17。 注意:传统上，前十分钟的过程中的AUC被认为是acutÈ胰岛素响应于葡萄糖（ 空气 ），而从过程的最后20分钟的AUC被认为是晚期胰岛素反应（LIR）。当动物变得更加dysmetabolic，胰岛素曲线下面积将增加，这反映补偿增加胰岛素不敏感。作为动物转移到明显的糖尿病，然而，AUC将降低，经常最初在急性胰岛素相，由AIR捕获。 </li></ol>

<ol>
	<li>Bergman, R., Phillips, L., Cobelli, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Physiologic+evaluation+of+factors+controlling+glucose+tolerance+in+man.">Physiologic evaluation of factors controlling glucose tolerance in man.</a> J. Clin. Invest. 68, 1456-1457 (1981).</li><li>Bergman, R., Prager, R., Volund, A., Olefsky, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Equivalence+of+the+insulin+sensitivity+index+in+man+derived+by+the+minimal+model+and+the+euglycemic+glucose+clamp.">Equivalence of the insulin sensitivity index in man derived by the minimal model and the euglycemic glucose clamp.</a> J. Clin. Invest. 79, 790-800 (1987).</li><li>Hovorka, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Partitioning+glucose+distribution/transport,+disposal,+and+endogenous+production+during+IVGTT.">Partitioning glucose distribution/transport, disposal, and endogenous production during IVGTT.</a> Am. J. Physiol. Endocrinol. Metab. 282, E992-E1007 (2002).</li><li>Salinari, S., Guidone, C., Bertuzzi, A., Manco, M., Asnaghi, S., Mingrone, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=First-phase+insulin+secretion+restoration+and+differential+response+to+glucose+load+depending+on+the+route+of+administration+in+type+2+diabetic+subjects+after+beriatric+surgery.">First-phase insulin secretion restoration and differential response to glucose load depending on the route of administration in type 2 diabetic subjects after beriatric surgery.</a> Diabetes Care. 32 (3), 375-380 (2009).</li><li>Roden, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Clinical Diabetes Research: Methods and Techniques. , (2007).</li><li>Cobelli, C., Pacini, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insulin+secretion+and+hepatic+extraction+in+humans+by+minimal+modeling+of+c-peptide+and+insulin+kinetics.">Insulin secretion and hepatic extraction in humans by minimal modeling of c-peptide and insulin kinetics.</a> Diabetes. 37, 223-231 (1988).</li><li>Lorenzo, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disposition+index,+glucose+effectiveness,+and+conversion+to+type+2+diabetes:+the+insulin+resistance+atherosclerosis+study.">Disposition index, glucose effectiveness, and conversion to type 2 diabetes: the insulin resistance atherosclerosis study.</a> Diabetes Care. 33, 2098-2103 (2010).</li><li>Hansen, B. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Investigation+and+treatment+of+type+2+diabetes+in+nonhuman+primates.">Investigation and treatment of type 2 diabetes in nonhuman primates.</a> Methods Mol Biol. 933, 177-185 (2012).</li><li>Hansen, B. C., Bodkin, N. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Standardization+of+IVGTT.+Importance+of+method+used+to+calculate+glucose+disappearance.">Standardization of IVGTT. Importance of method used to calculate glucose disappearance.</a> Diabetes Care. 16 (5), 847 (1993).</li><li>Hardwood, J. H., Listrani, P., Wagner, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nonhuman+primates+and+other+animal+models+in+diabetes+research.">Nonhuman primates and other animal models in diabetes research.</a> J Diabetes Sci Tech. 3, 503-514 (2012).</li><li>De Koning, E. J., Bodkin, N. L., Hansen, B. C., Clark, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diabetes+mellitus+in+Macaca+mulatta+monkeys+is+characterized+by+islet+amyloidosis+and+reduction+in+beta-cell+population.">Diabetes mellitus in Macaca mulatta monkeys is characterized by islet amyloidosis and reduction in beta-cell population.</a> Diabetologia. 36, 378-384 (1993).</li><li>Letiexhe, M. R., Scheen, A. J., Gerard, P. L., Desaive, C., Lefebvre, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Insulin+secretion,+clearance+and+action+before+and+after+gastroplasty+in+severely+obese+subjects.">Insulin secretion, clearance and action before and after gastroplasty in severely obese subjects.</a> Int J Obes Relat Metab Disord. 18, 295-300 (1994).</li><li>Letiexhe, M. R., Scheen, A. J., Gerard, P. L., Desaive, C., Lefebvre, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Postgastroplasty+recovery+of+ideal+body+weight+normalizes+glucose+and+insulin+metabolism+in+obese+women.">Postgastroplasty recovery of ideal body weight normalizes glucose and insulin metabolism in obese women.</a> J Clin Endocrinol Metab. 80, 364-369 (1995).</li><li>Kim, S. H., Abbasi, F., Chu, J. W., McLaughlin, T. L., Lamendola, C., Polonsky, K. S., Reaven, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rosiglitazone+reduces+glucose-stimulated+insulin+secretion+rate+and+increases+insulin+clearance+in+nondiabetic,+insulin-resistant+individuals.">Rosiglitazone reduces glucose-stimulated insulin secretion rate and increases insulin clearance in nondiabetic, insulin-resistant individuals.</a> Diabetes. 54, 2447-2452 (2005).</li><li>Toffolo, G., Breda, E., Cavaghan, M. K., Ehrmann, D. A., Polonsky, K. S., Cobelli, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+indexes+of+beta-cell+function+during+graded+up+and+down+glucose+infusion+from+C-peptide+minimal+models.">Quantitative indexes of beta-cell function during graded up and down glucose infusion from C-peptide minimal models.</a> Am J Physiol Endocrinol Metab. 280, E2-E10 (2001).</li><li>Wang, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantification+of+beta-cell+insulin+secretory+function+using+a+graded+glucose-infusion+with+C-peptide+deconvolution+in+dysmetabolic,+and+diabetic+cynomolgus+monkeys.">Quantification of beta-cell insulin secretory function using a graded glucose-infusion with C-peptide deconvolution in dysmetabolic, and diabetic cynomolgus monkeys.</a> Diabetology and Metabolic Syn. 5, 40 (2013).</li><li>Xiao, Y. F., Wang, B., Wang, X., Du, F., Benzinou, M., Wang, Y. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Xylazine-induced+reduction+of+tissue+sensitivity+to+insulin+leads+to+acute+hyperglycemia+in+diabetic+and+normoglycemic+monkeys.">Xylazine-induced reduction of tissue sensitivity to insulin leads to acute hyperglycemia in diabetic and normoglycemic monkeys.</a> Anesthesiology. 13 (33), (2013).</li><li>Porte, D., Kahn, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=&#946;-cell+dysfunction+and+failure+in+type+2+diabetes+potential+mechanisms.">&#946;-cell dysfunction and failure in type 2 diabetes potential mechanisms.</a> Diabetes. 50, S160-S163 (2001).</li><li>DeFronzo, R. A., Tobin, J. D., Andres, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glucose+clamp+technique:+a+method+for+quantifying+insulin+secretion+and+resistance.">Glucose clamp technique: a method for quantifying insulin secretion and resistance.</a> American Journal of Physiology. 237 (3), G214-G223 (1979).</li><li>Ferrannini, E., Gastaldelli, A., Miyazaki, Y., Matsuda, M., Mari, A., DeFronzo, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=&#946;-cell+function+in+subjects+spanning+the+range+from+normal+glucose+tolerance+to+overt+diabetes:+a+new+analysis.">&#946;-cell function in subjects spanning the range from normal glucose tolerance to overt diabetes: a new analysis.</a> J Clin Endocrinol Metab. 90 (1), 493-500 (2005).</li><li>Vaughan, K. L., Szarowicz, M. D., Herbert, R. L., Mattison, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+anesthesia+protocols+for+intravenous+glucose+tolerance+testing+in+rhesus+monkeys.">Comparison of anesthesia protocols for intravenous glucose tolerance testing in rhesus monkeys.</a> J Med Primatol. 43, 162-168 (2014).</li><li>Kemnitz, J. W., Kraemer, G. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+glucoregulation+in+rhesus+monkeys+sedated+with+ketamine.">Assessment of glucoregulation in rhesus monkeys sedated with ketamine.</a> American Journal of Primatology. 3, 201-210 (1982).</li><li>Dutton, C. J., Parvin, C. A., Gronowski, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+glycated+hemoglobin+percentages+for+use+in+the+diagnosis+and+monitoring+of+diabetes+mellitus+in+nonhuman+primates.">Measurement of glycated hemoglobin percentages for use in the diagnosis and monitoring of diabetes mellitus in nonhuman primates.</a> Am J Vet Res. 64, 562-568 (2003).</li><li>Rai, V., Iyer, U., Mani, I., Mani, U. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Serum+biochemical+changes+in+insulin+dependent+and+non-insulin+dependent+diabetes+mellitus+and+their+role+in+the+development+of+secondary+complications.">Serum biochemical changes in insulin dependent and non-insulin dependent diabetes mellitus and their role in the development of secondary complications.</a> Int J Diab Dev Countries. 17, 33-37 (1997).</li><li>Shirasaki, Y., Yoshioka, N., Kanazawa, K., Maekawa, T., Horikawa, T., Hayashi, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+physical+restraint+on+glucose+tolerance+in+cynomolgus+monkeys.">Effect of physical restraint on glucose tolerance in cynomolgus monkeys.</a> J Med Primatol. 42, 165-168 (2013).</li></ol>

作者是附属于合同研究组织（冠科生物）活跃在代谢性疾病领域。

的IVGTT评估葡萄糖刺激的胰岛素释放的基于体重5，12，13的单一的葡萄糖输注的能力。从测定中，空腹血糖和胰岛素水平达到，并且它允许动物的能力来评估释放胰岛素和朝基线返回升高的血糖水平。这提供了信息来表征动物作为一个正常的血糖和胰岛素水平健康对照，具有正常血糖高胰岛素dysmetabolic动物，或高血糖的胰岛素抵抗性糖尿病动物的用户。 重要的是血液?...

的IVGTT是常规用于确定在人类中的β细胞功能在不同的代谢状态5,7的便利功能测定法。在T2D中的动物模型中，它被公认为表征的动物，显示从代谢疾病进展的工具一个健康的一个dysmetabolic高血糖状态8,9。T2D的最接近动物模型是表现在非人灵长类动物（NHPS），其中恒河猴和食蟹猴是显着的例子。这些动物自然发展随着年龄和肥胖促进其发生率相同的风险因素，如2型糖尿病的人类10。此外，还有一个类似的疾病进展和胰腺病理表示作为dysmetabolic疾病进展11淀粉样蛋白沉积物。 在这里，我们对我们在执行的NHPS作为IVGTT我们对这些动物代谢状态的殖民地特征的一部分的标准方法报告。此方法是容易进行相对于其它更耗时且昂贵的技术2。该IVGTT是一个用于快速和频繁的表征大群动物是有用的。当与糖化血红蛋白（HbA1c）的电平考虑在内的动物的饮食和食物摄取的历史，以及他们的百分之瘦体重和体脂肪，所述IVGTT通常足以用于向显性糖尿病6表征动物的代谢状态和进展8。 糖化血红蛋白代表了一个红血细胞的寿命的平均血糖水平，过去6周〜3个月，只要葡萄糖水平的可靠量度。当从IVGTT的禁食基线血样测定，该值在提供程序之间的几个月一个窗口，血糖控制。如果动物已经从dysmetabolic转变自上次IVGTT糖尿病，比前值高得多的糖化血红蛋白数值表明该过渡即将开始他们的最后IVGTT后，而，糖化血红蛋白值更接近其先前的数值表明，他们最近才转移。在一般情况下，在恒河猴，糖化血红蛋白值大于6％的被认为是不正常的，并指示血糖控制不佳10,23。 血糖水平应的动物作为一个整体的行为和一般健康状况的范围内进行解释。糖尿病猕猴 - 人一样 - 展览亢进，多饮，多尿。动物群的住房提供了这些指标和个人护理dysmetabolic和糖尿病的猴子所需的测量显著的挑战。我们建议单独容纳动物，以便可以提供更个性化的服务，并且更容易的猴的健康行为标记物来监测8。此外，糖尿病猕猴将表现出体重下降，以及升高的脂质分布（增加的胆固醇，高甘油三酯血症）和血清化学干扰矿物质代谢。它测量肝脏和血清化学肾功能的标志物是重要的，因为这些器官损伤往往前进代谢紊乱/糖尿病的并发症，并且可以是血糖，脂质和矿物质失衡9，11，18的共同决定因素，24 。 使用此方法时，来自多个，经常刻画产生的历史价值在猴子的生活中具有特殊的价值。如果其他程序，如葡萄糖钳夹或分级葡萄糖输注（GGI），需要充分评估动物的健康，这是常用于初步鉴定时，他们的历史是不可用。每三个月的频率。然而，一旦一个基线已经建立，重复IVGTTs通常足以跟踪动物的进展。当动物被注册到整个多项研究，这是特别重要历年的代谢状态为主。而他们的健康可能在一个时间数年保持相对稳定，当动物的代谢状况恶化，在胰岛素抵抗和葡萄糖不耐症的显着增加可以非常迅速地发生。糖化血红蛋白值允许的预定相隔3个月的程序之间的动物的健康状况的下降或改善一些插值。由于这个原因，这种方法是理想的特征在多个纵向研究对其自然寿命的过程中使用的动物。

<table><tbody><tr><td>Allegra X-15R Centrifuge</td><td></td><td></td><td>plasma: 4 &amp;#176;C at 3,000 rpm for 10 min</td></tr><tr><td>Sorvall ST16R Centrifuge</td><td></td><td></td><td>serum: 22 &amp;#176;C at 3,000 rpm for 10 min</td></tr><tr><td>Thermo Scientific -86 &amp;#176;C Freezer, Forma 88000 Series</td><td></td><td>Model: 88500A</td><td></td></tr><tr><td>Dextrose 50% (D50)</td><td>Webster</td><td>07-8008986</td><td>I.V. glucose infusate</td></tr><tr><td>3 ml Luer Lock Syringe</td><td>Midwest Veterinary Suppy</td><td></td><td>serial blood draws</td></tr><tr><td>5 ml Luer Lock Syringe</td><td>Midwest Veterinary Suppy</td><td></td><td>heparinized saline flush</td></tr><tr><td>10 ml Luer Lock Syringe</td><td>Midwest Veterinary Suppy</td><td></td><td>delivery of I.V. D50</td></tr><tr><td>Gauze sponges 2 x 2</td><td>Midwest Veterinary Suppy</td><td>366.23000.4</td><td>Used Dry, w/ 70% Alcohol, and 2% Chlorohex Solution</td></tr><tr><td>4 ml serum separator tubes&amp;nbsp;</td><td>Midwest Veterinary Supply</td><td>366.45000.4</td><td>blood collection tube for superchem panel</td></tr><tr><td>K&lt;sub&gt;2&lt;/sub&gt;EDTA, 2 ml</td><td>VWR</td><td>95057-239</td><td>blood collection tubes</td></tr><tr><td>Aprotinin, 100 mg</td><td>Sigma</td><td>A1153-100MG</td><td>blood collection tube protease additive</td></tr><tr><td>22 G x 1&quot; Catheters</td><td>Midwest Veterinary Suppy</td><td>193.75250.2</td><td>I.V. catheter&amp;nbsp;</td></tr><tr><td>Injection Plug W/ Cap</td><td>Midwest Veterinary Suppy</td><td>001.11500.2</td><td>%50 dextrose infusion port</td></tr><tr><td>Porus Tape, 1/2&quot; x 10 yd</td><td>Midwest Veterinary Suppy</td><td>001.85000.2</td><td>maintain adherance of catheters and hep. Locks</td></tr><tr><td>Chlorhexidine Solution 2%</td><td>Midwest Veterinary Suppy</td><td>193.08855.3</td><td>prep catheter site</td></tr><tr><td>70% Ethanol</td><td>VWR</td><td>71001-654</td><td>prep catheter site</td></tr><tr><td>tourniquet</td><td>Webster</td><td>07-8003432</td><td></td></tr><tr><td>3-way stopcock</td><td>Midwest Veterinary Supply</td><td>366.28510.4</td><td>hep. lock</td></tr><tr><td>37&quot; extension set</td><td>Webster</td><td>07-8454200</td><td>hep. lock</td></tr><tr><td>Exel 50-60cc LL Syringes</td><td>Midwest Veterinary Suppy</td><td>001.12250.2</td><td>Heparinized saline flush</td></tr><tr><td>250 ml bag 0.9% saline</td><td>Webster</td><td>07-8365593</td><td>flush</td></tr><tr><td>1,000 U Heparin, 10 ml</td><td>Webster</td><td>07-883-4916</td><td></td></tr><tr><td>Ketamine (Ketaset) 100&amp;nbsp;mg/ml</td><td>Fort Dodge</td><td>(AV ordered)</td><td></td></tr><tr><td>Precision Xtra glucose test strips 50/bx</td><td>Abbott (American Diabetes Wholesale)</td><td>9381599728K7</td><td>test baseline/T3 blood glucose levels</td></tr><tr><td>Masimo Rad 57</td><td>DRE</td><td>6052057V</td><td>pulse-oximeter</td></tr><tr><td>Pavia rectal thermometer</td><td>Patterson</td><td>07-8391335</td><td></td></tr><tr><td>Precision Xtra Glucometer</td><td>Abbott</td><td>9381599728K7</td><td>Handheld glucometer</td></tr></tbody></table>

characterization of metabolic status in nonhuman primates with the intravenous glucose tolerance test

静脉内葡萄糖耐量试验（IVGTT）起着葡萄糖稳态的表征了关键作用。当与血清生化型材，包括在这两个馈血糖水平和禁食状态，糖化血红蛋白，胰岛素水平，饮食的临床病史，身体组成和体重状态，正常和异常的血糖控制的评估一起可以制成。一个IVGTT的解释是通过在葡萄糖和胰岛素水平的变化的测量随时间的关系做了葡萄糖挑战。要考虑的关键组成部分是:在关系达到T0（葡萄糖输注结束），葡萄糖清除率ķ从快速葡萄糖清除的斜率在第20分钟，衍生（T1至T20），时间峰的葡萄糖和胰岛素水平返回到葡萄糖基线和曲线（AUC）下的面积。这些IVGTT措施将显示来自健康t特性变化的葡萄糖稳态举动OA患病代谢状态5。这里我们将描述非人灵长类动物（猕猴和食蟹猕猴），这是在人类中II型糖尿病（T2D）的最相关的动物模型和IVGTT和这些动物的临床概况从表征贫健康，肥胖dysmetabolic，和T2D状态8，10，11。

在图1中所示的结果是示范从成熟，健康和糖尿病猕猴在30分钟IVGTT的过程中典型的葡萄糖和胰岛素的曲线。从健康和晚期糖尿病猴子数据示为了对比，从代谢的表征的范围的两个极端处的动物之间的明显的差异。此IVGTT协议已经通过在具有类似的结果恒河猴作者成功使用。 一个健康的猕猴（恒河猴和食蟹猴）的禁?...

Watch this Scientific Journal Video about Characterization of Metabolic Status in Nonhuman Primates with the Intravenous Glucose Tolerance Test at JoVE.com

Characterization of Metabolic Status in Nonhuman Primates with the Intravenous Glucose Tolerance Test

该协议的目的是提出一种标准方法来执行静脉内葡萄糖耐量试验（IVGTTs）评估在非人灵长类动物的血糖控制和从健康评估dysmetabolic其代谢状态。

代谢状态的表征非人灵长类动物与静脉葡萄糖耐量试验

The intravenous glucose tolerance test (IVGTT) plays a key role in the characterization of glucose homeostasis. When taken together with serum biochemical ...

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Research

JoVE Journal

Medicine

10.8K 次观看.  Crown Bioscience. 该协议的目的是提出一种标准方法来执行静脉内葡萄糖耐量试验（IVGTTs）评估在非人灵长类动物的血糖控制和从健康评估dysmetabolic其代谢状态。 

视频: Characterization of Metabolic Status in Nonhuman Primates with the Intravenous Glucose Tolerance Test

Analysis of Non-Human Primate Pancreatic Islet Oxygen Consumption

The measurement of oxygen consumption in spheroid clusters of cells, such as ex vivo pancreatic islets, has historically been challenging. We demonstrate the measurement of islet oxygen consumption using a 96-well microplate designed for the measurement of oxygen consumption in spheroids. In this assay, spheroid microplates are coated with a cell and tissue adhesive on the day prior to the assay. We utilize a small volume of adhesive solution to encourage islet adherence to only the bottom of the well. On the day of the assay, 15 islets are loaded directly into the base of each well using a technique that ensures optimal positioning of islets and accurate measurement of oxygen consumption. Various aspects of mitochondrial respiration are probed pharmacologically in non-human primate islets, including ATP-dependent respiration, maximal respiration, and proton leak. This method allows for consistent, reproducible results using only a small number of islets per well. It can theoretically be applied to any cultured spheroids of similar size.

This protocol demonstrates the accurate and reproducible measurement of oxygen consumption in non-human primate pancreatic islets. The islet loading techniques and coating of the microplate provide a framework for efficient measurement of respiration in other types of cultured spheroids.

非人类灵长类动物胰腺胰岛耗氧量分析

The measurement of oxygen consumption in spheroid clusters of cells, such as ex vivo pancreatic islets, has historically been challenging. We demonstrate ...

科研

发育生物学

Intraperitoneal Glucose Tolerance Test, Measurement of Lung Function, and Fixation of the Lung to Study the Impact of Obesity and Impaired Metabolism on Pulmonary Outcomes

Obesity and respiratory disorders are major health problems. Obesity is becoming an emerging epidemic with an expected number of over 1 billion obese individuals worldwide by 2030, thus representing a growing socioeconomic burden. Simultaneously, obesity-related comorbidities, including diabetes as well as heart and chronic lung diseases, are continuously on the rise. Although obesity has been associated with increased risk for asthma exacerbations, worsening of respiratory symptoms, and poor control, the functional role of obesity and perturbed metabolism in the pathogenesis of chronic lung disease is often underestimated, and underlying molecular mechanisms remain elusive. This article aims to present methods to assess the effect of obesity on metabolism, as well as lung structure and function. Here, we describe three techniques for mice studies: (1) assessment of intraperitoneal glucose tolerance (ipGTT) to analyze the effect of obesity on glucose metabolism; (2) measurement of airway resistance (Res) and respiratory system compliance (Cdyn) to analyze the effect of obesity on lung function; and (3) preparation and fixation of the lung for subsequent quantitative histological assessment. Obesity-related lung diseases are probably multifactorial, stemming from systemic inflammatory and metabolic dysregulation that potentially adversely influence lung function and the response to therapy. Therefore, a standardized methodology to study molecular mechanisms and the effect of novel treatments is essential.

The incidence of obesity is rising and increases the risk of chronic lung diseases. To establish the underlying mechanisms and preventive strategies, well-defined animal models are needed. Here, we provide three methods (glucose-tolerance-test, body plethysmography, and lung fixation) to study the effect of obesity on pulmonary outcomes in mice.

腹腔葡萄糖耐受试验、肺功能测定和肺固定研究肥胖和代谢障碍对肺部结局的影响

Obesity and respiratory disorders are major health problems. Obesity is becoming an emerging epidemic with an expected number of over 1 billion obese ...

免疫与感染

Determining Glucose Metabolism Kinetics Using 18F-FDG Micro-PET/CT

This paper describes the use of 18F-FDG and micro-PET/CT imaging to determine in vivo glucose metabolism kinetics in mice (and is transferable to rats). Impaired uptake and metabolism of glucose in multiple organ systems due to insulin resistance is a hallmark of type 2 diabetes. The ability of this technique to extract an image-derived input function from the vena cava using an iterative deconvolution method eliminates the requirement of the collection of arterial blood samples. Fitting of tissue and vena cava time activity curves to a two-tissue, three compartment model permits the estimation of kinetic micro-parameters related to the 18F-FDG uptake from the plasma to the intracellular space, the rate of transport from intracellular space to plasma and the rate of 18F-FDG phosphorylation. This methodology allows for multiple measures of glucose uptake and metabolism kinetics in the context of longitudinal studies and also provides insights into the efficacy of therapeutic interventions.

Determining Glucose Metabolism Kinetics Using 18F-FDG Micro-PET/CT

This study describes a protocol that uses 18F-FDG and positron emission tomography/computed tomography (PET/CT) imaging, together with kinetic modelling, to quantify the in vivo, real-time uptake of 18F-FDG into tissues.

确定葡萄糖代谢动力学研究<sup&gt; 18</sup&gt; F-FDG微PET / CT

This paper describes the use of 18F-FDG and micro-PET/CT imaging to determine in vivo glucose metabolism kinetics in mice (and is transferable to rats). ...

医学

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

Obesity represents the most important single risk factor in the pathogenesis of type 2 diabetes, a disease which is characterized by a resistance to insulin-stimulated glucose uptake and a gross decompensation of systemic glucose metabolism. Despite considerable progress in the understanding of glucose metabolism, the molecular mechanisms of its regulation in health and disease remain under-investigated, while novel approaches to prevent and treat diabetes are urgently needed. Diet derived glucose stimulates the pancreatic secretion of insulin, which serves as the principal regulator of cellular anabolic processes during the fed-state and thus balances blood glucose levels to maintain systemic energy status. Chronic overfeeding triggers meta-inflammation, which leads to alterations in peripheral insulin receptor-associated signaling and thus reduces the sensitivity to insulin-mediated glucose disposal. These events ultimately result in elevated fasting glucose and insulin levels as well as a reduction in glucose tolerance, which in turn serve as important indicators of insulin resistance. Here, we present a protocol for the generation and metabolic characterization of high-fat diet (HFD)-fed mice as a frequently used model of diet-induced insulin resistance. We illustrate in detail the oral glucose tolerance test (OGTT), which monitors the peripheral disposal of an orally administered glucose load and insulin secretion over time. Additionally, we present a protocol for the insulin tolerance test (ITT) to monitor whole-body insulin action. Together, these methods and their downstream applications represent powerful tools to characterize the general metabolic phenotype of mice as well as to specifically assess alterations in glucose metabolism. They may be especially useful in the broad research field of insulin resistance, diabetes and obesity to provide a better understanding of pathogenesis as well as to test the effects of therapeutic interventions.

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

The current article describes the generation and metabolic characterization of high-fat diet-fed mice as a model of diet-induced insulin resistance and obesity. It further features detailed protocols to perform the oral glucose tolerance test and the insulin tolerance test, monitoring whole-body alterations of glucose metabolism in vivo.

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)

Obesity represents the most important single risk factor in the pathogenesis of type 2 diabetes, a disease which is characterized by a resistance to ...

Body Composition and Metabolic Caging Analysis in High Fat Fed Mice

Alterations to body composition (fat or lean mass), metabolic parameters such as whole-body oxygen consumption, energy expenditure, and substrate utilization, and behaviors such as food intake and physical activity can provide important information regarding the underlying mechanisms of disease. Given the importance of body composition and metabolism to the development of obesity and its subsequent sequelae, it is necessary to make accurate measures of these parameters in the pre-clinical research setting. Advances in technology over the past few decades have made it possible to derive these measures in rodent models in a non-invasive and longitudinal fashion. Consequently, these metabolic measures have proven useful when assessing the response of genetic manipulations (for example knockout or transgenic mice, viral knock-down or overexpression of genes), experimental drug/compound screening and dietary, behavioral or physical activity interventions. Herein, we describe the protocols used to measure body composition and metabolic parameters using an animal monitoring system in chow-fed and high fat diet-fed mice.

This protocol describes the use of a body composition analyzer and metabolic animal monitoring system to characterize body composition and metabolic parameters in mice. An obesity model induced by high-fat feeding is used as an example for the application of these techniques.

高脂喂养小鼠体内成分及代谢锁定分析

Alterations to body composition (fat or lean mass), metabolic parameters such as whole-body oxygen consumption, energy expenditure, and substrate ...

Acclimation Prior to an Intraperitoneal Insulin Tolerance Test to Mitigate Stress-Induced Hyperglycemia in Conscious Mice

The insulin tolerance test is commonly used in metabolic studies to assess whole body insulin sensitivity in rodents. It is a relatively simple test that involves measurement of blood glucose levels over time following a single intraperitoneal injection of insulin. Given that it is performed in the conscious state and blood is often collected via a tail snip, it has the potential to elicit a stress response from animals due to anxiety associated with handling and blood collection. As such, a stress-induced rise in blood glucose can occur, making it difficult to detect and interpret the primary endpoint measure, namely an insulin-mediated reduction in blood glucose. This has been seen in many mouse strains, and is quite common in diabetic db/db mice, where glucose levels can increase, rather than decrease, after insulin administration. Here, we describe a method of acclimating mice to handling, injections and blood sampling prior to performing the insulin tolerance test. We find that this lowers stress-induced hyperglycemia and results in data that more accurately reflects whole body insulin sensitivity.

Stress-induced elevations in glucose levels can confound interpretation of data derived from a conscious intraperitoneal insulin tolerance test in mice. In this article, we describe a method to acclimate the mice to handling, injections and blood sampling prior to performing the insulin tolerance test in order to limit stress-induced hyperglycemia.

The insulin tolerance test is commonly used in metabolic studies to assess whole body insulin sensitivity in rodents. It is a relatively simple test that ...

Evaluation of Hepatic Glucose Production in a Polycystic Ovary Syndrome Mouse Model

Polycystic ovary syndrome (PCOS) is a common disease that results in disorders of glucose metabolism, such as insulin resistance and glucose intolerance. Dysregulated glucose metabolism is an important manifestation of the disease and is the key to its pathogenesis. Therefore, studies involving evaluation of glucose metabolism in PCOS are of utmost importance. Very few studies have quantified hepatic glucose production directly in PCOS models using non-radioactive glucose tracers. In this study, we discuss step-by-step instructions for the quantification of the rate of hepatic glucose production in a PCOS mouse model by measuring M+2 enrichment of [6,6-2H2]glucose, a stable isotopic glucose tracer, via gas chromatography - mass spectrometry (GCMS). This procedure involves creation of stable isotopic glucose tracer solution, use of tail vein catheter placement and infusion of the glucose tracer in both fasting and glucose-rich states in the same mouse in tandem. The enrichment of [6,6-2H2]glucose is measured using pentaacetate derivative in GCMS. This technique can be applied to a wide variety of studies involving direct measurement of the rate of hepatic glucose production.

This study describes the direct measurement of hepatic glucose production in a polycystic ovary syndrome mouse model by using a stable isotopic glucose tracer via tail vein in both fasting and glucose-rich states in tandem.

多囊卵巢综合征小鼠模型中肝葡萄糖产生的评价

Polycystic ovary syndrome (PCOS) is a common disease that results in disorders of glucose metabolism, such as insulin resistance and glucose intolerance. ...

Glucose Uptake Measurement and Response to Insulin Stimulation in In Vitro Cultured Human Primary Myotubes

Skeletal muscle is the largest glucose deposit in mammals and largely contributes to glucose homeostasis. Assessment of insulin sensitivity of muscle cells is of major relevance for all studies dedicated to exploring muscle glucose metabolism and characterizing metabolic alterations. In muscle cells, glucose transporter type 4 (GLUT4) proteins translocate to the plasma membrane in response to insulin, thus allowing massive entry of glucose into the cell. The ability of muscle cells to respond to insulin by increasing the rate of glucose uptake is one of the standard readouts to quantify muscle cell sensitivity to insulin. Human primary myotubes are a suitable in vitro model, as the cells maintain many features of the donor phenotype, including insulin sensitivity. This in vitro model is also suitable for the test of any compounds that could impact insulin responsiveness. Measurements of the glucose uptake rate in differentiated myotubes reflect insulin sensitivity.
In this method, human primary muscle cells are cultured in vitro to obtain differentiated myotubes, and glucose uptake rates with and without insulin stimulation are measured. We provide a detailed protocol to quantify passive and active glucose transport rates using radiolabeled [3H] 2-deoxy-D-Glucose ([3H]2dG). Calculation methods are provided to quantify active basal and insulin-stimulated rates, as well as stimulation fold.

Glucose Uptake Measurement and Response to Insulin Stimulation in In Vitro Cultured Human Primary Myotubes

In this method, human primary muscle cells are cultured in vitro to obtain differentiated myotubes and glucose uptake rates are measured. We provide a detailed protocol to quantify rates in basal and insulin-stimulated states using radiolabeled [3H] 2-deoxy-D-Glucose.

葡萄糖摄取测量和对胰岛素刺激的反应<em&gt;体外</em&gt;培养的人类初级肌管

Skeletal muscle is the largest glucose deposit in mammals and largely contributes to glucose homeostasis. Assessment of insulin sensitivity of muscle ...

生物学

Using a Combination of Indirect Calorimetry, Infrared Thermography, and Blood Glucose Levels to Measure Brown Adipose Tissue Thermogenesis in Humans

In mammals, brown adipose tissue (BAT) is activated rapidly in response to cold in order to maintain body temperature. Although BAT has been studied greatly in small animals, it is difficult to measure the activity of BAT in humans. Therefore, little is known about the heat-generating capacity and physiological significance of BAT in humans, including the degree to which components of the diet can activate BAT. This is due to the limitations in the currently most used method to assess the activation of BAT-radiolabeled glucose (fluorodeoxyglucose or 18FDG) measured by positron emission tomography-computerized tomography (PET-CT).
This method is usually performed in fasted subjects, as feeding induces glucose uptake by the muscles, which can mask the glucose uptake into the BAT. This paper describes a detailed protocol for quantifying total-body human energy expenditure and substrate utilization from BAT thermogenesis by combining indirect calorimetry, infrared thermography, and blood glucose monitoring in carbohydrate-loaded adult males. To characterize the physiological significance of BAT, measures of the impact of BAT activity on human health are critical. We demonstrate a protocol to achieve this by combining carbohydrate loading and indirect calorimetry with measurements of supraclavicular changes in temperature. This novel approach will help to understand the physiology and pharmacology of BAT thermogenesis in humans.

Here, we present a protocol to quantify the physiological significance of the impact of brown adipose tissue (BAT) activity on human metabolism. This is achieved by combining carbohydrate loading and indirect calorimetry with measurements of supraclavicular changes in temperature. This novel approach can help develop a pharmacological target for BAT thermogenesis in humans.

使用间接量热法、红外热成像和血糖水平的组合来测量人类棕色脂肪组织的产热

In mammals, brown adipose tissue (BAT) is activated rapidly in response to cold in order to maintain body temperature. Although BAT has been studied ...

Assessment of the Metabolic Effects of Isocaloric 2:1 Intermittent Fasting in Mice

Intermittent fasting (IF), a dietary intervention involving periodic energy restriction, has been considered to provide numerous benefits and counteract metabolic abnormalities. So far, different types of IF models with varying durations of fasting and feeding periods have been documented. However, interpreting the outcomes is challenging, as many of these models involve multifactorial contributions from both time- and calorie-restriction strategies. For example, the alternate day fasting model, often used as a rodent IF regimen, can result in underfeeding, suggesting that health benefits from this intervention are likely mediated via both caloric restriction and fasting-refeeding cycles. Recently, it has been successfully demonstrated that 2:1 IF, comprising 1 day of fasting followed by 2 days of feeding, can provide protection against diet-induced obesity and metabolic improvements without a reduction in overall caloric intake. Presented here is a protocol of this isocaloric 2:1 IF intervention in mice. Also described is a pair-feeding (PF) protocol required to examine a mouse model with altered eating behaviors, such as hyperphagia. Using the 2:1 IF regimen, it is demonstrated that isocaloric IF leads to reduced body weight gain, improved glucose homeostasis, and elevated energy expenditure. Thus, this regimen may be useful to investigate the health impacts of IF on various disease conditions.

The current article describes a detailed protocol for isocaloric 2:1 intermittent fasting to protect and treat against obesity and impaired glucose metabolism in wild-type and ob/ob mice.

小鼠间间间禁食的代谢效应评估 2:1 间歇禁食

Intermittent fasting (IF), a dietary intervention involving periodic energy restriction, has been considered to provide numerous benefits and counteract ...

代谢状态的表征非人灵长类动物与静脉葡萄糖耐量试验

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