扩展高胰岛素-正葡萄糖钳夹技术的建立

目的建立扩展高胰岛素-正葡萄糖钳夹技术.方法应用扩展葡萄糖钳夹技术(即联合葡萄糖钳夹、3-3H标记葡萄糖示踪技术及间接测热技术)对9例正常体重、正常糖耐量者进行方法学研究.结果(1)在优势浓度胰岛素、血浆葡萄糖稳定在正常水平状态下肝糖产生完全被抑制,未见到升糖激素(皮质醇、生长激素、胰高血糖素)显著兴奋及内源性胰岛素释放.(2)扩展高胰岛素-正葡萄糖钳夹稳态期,胰岛素介导的机体葡萄糖利用率(葡萄糖消失率)较基础状态显著增加[(5.86±0.65)mg*kg-1*min-1比(2.45±0.15)mg*kg-1*min-1,P＜0.01],并以糖原合成为主;脂肪氧化被显著抑制,游离脂肪酸浓度显著下降.结论扩展高胰岛素-正葡萄糖钳夹技术已成功建立;在外源性胰岛素-葡萄糖代谢的稳定状态下,机体对葡萄糖利用显著增加,脂肪分解显著减少.     

应用高胰岛素正葡萄糖钳夹技术评价我国LADA患者的胰岛素抵抗

目的应用高胰岛素正葡萄糖钳夹技术评估我国成人隐匿性自身免疫性糖尿病（LADA）患者的胰岛素抵抗。方法研究对象分为LADA组、2型糖尿病（T2DM）组、正常对照（NC）组。糖尿病患者均为初诊未治,经2周胰岛素强化治疗血糖控制达标后进行研究。对全部研究对象空腹测定临床参数及生化指标,应用高胰岛素正葡萄糖钳夹技术测定胰岛素敏感性指数（ISI）。结果LADA组FC-P及Fins显著低于NC组（P〈0．05）,年龄、BMI、wHR、SBP、DBP、TC、TG、LDL—C、HDL-C与NC组比较,均无统计学差异。LADA组BMI、WHR、SBP、Fins、FC-P、TC、TG、LDL-C均显著低于T2DM组（P均〈0．05）。NC组、LADA组、T2DM组ISI分别为12．83±1．09、6．70±0．71、3．80±0．20,三组间比较有统计学差异（P〈0．05）。结论与NC组相比,LADA患者存在胰岛素抵抗,其程度较T2DM组轻。     

两步法高胰岛素-正常葡萄糖钳夹技术的建立

目的　建立两步法高胰岛素 正常葡萄糖钳夹技术。　方法　应用两步法高胰岛素-正常葡萄糖钳夹术(第一阶段胰岛素输注速率为 30 mU·m-2 ·min-1,第二阶段为 120 mU·m-2 ·min-1),评估10名健康女性志愿者的胰岛素敏感性,同时监测钳夹试验过程中循环血内源性升糖激素(胰高血糖素、皮质醇、生长激素)和游离脂肪酸(FFA)浓度变化。　结果　(1)两步法高胰岛素 正常葡萄糖钳夹术第一阶段(60~90 min)和第二阶段(150~180 min) 稳态血胰岛素分别维持在(46±7)mU/L和(218±27)mU/L,稳态葡萄糖利用率分别为(7.2±1.4)mg·kg-1·min-1和(13.1±1.2)mg·kg-1·min-1;(2)钳夹试验过程中,血清胰高血糖素、皮质醇和 FFA水平较基础值均有显著下降,而生长激素和C肽水平无明显改变。　结论　成功建立了两步法高胰岛素 正常葡萄糖钳夹术;钳夹试验过程中高浓度的胰岛素抑制了胰高血糖素、皮质醇的分泌,并减少脂肪分解。     

高葡萄糖钳夹技术的建立

目的　在国内建立检测胰岛素 (Ins)分泌功能的精确方法 :高葡萄糖钳制技术。　方法　对 2 5名居住上海地区的正常中国人 (男 16名 ,女 9名 )进行高葡萄糖钳夹实验。空腹平卧后两侧上肢静脉埋管。一侧持续输注 2 0 %葡萄糖溶液 ,通过负反馈调节机制使血糖始终维持在较基础血糖升高 7.9mmol/L的水平。另一侧静脉定时取血测定Ins水平 ,以获得双相Ins分泌值 ,评价胰岛 β细胞功能。　结果　 (1)受试者血浆葡萄糖浓度在钳夹开始后的指定时间内 (14min)能达到实验所要求的高糖平台 [(12 .86± 0 .11)mmol/L] ,并能维持此水平至实验结束 ;(2 )在维持高糖平台期间 ,每 5min检测的血糖变异系数为 4.2 % ;(3 )可观察到持续高糖水平下双相Ins的分泌形态 ,第一时相Ins分泌为 (2 47.8± 2 3 .5)mU/L ,第二时相为 (72 .8± 6.1)mU/L。 (4)高糖钳夹可同时获得个体的Ins敏感性 (ISI) 19.9± 2 .4,但较正糖钳夹值为高。　结论　 (1)成功建立了高葡萄糖钳夹技术 ,评价了正常个体的双相Ins分泌及最大Ins分泌。 (2 )高葡萄糖钳夹能同时了解机体的胰岛素抵抗 (IR)情况 ,反映不同人群ISI的差异     

葡萄糖钳夹技术在糖尿病研究中的应用

本文介绍葡萄糖钳夹技术在糖尿病研究中的应用。通过葡萄糖钳夹技术可研究在正常或高血糖情况下机体的胰岛素敏感性，胰岛素分泌和葡萄糖，脂类和蛋白质代谢等变化。     

不同糖耐量人群HOMA-IR与正常血糖高胰岛素钳夹试验的相关性分析

目的 评价各项胰岛素抵抗指标,尤其是稳态模型评估的胰岛素抵抗指数(HOMA-IR)与正常血糖高胰岛素钳夹试验间的相关性.方法 正常糖耐量(NGT)组、糖耐量受损(IGT)组和2型糖尿病组共142名受试者均接受体重、身高、腰围、空腹血糖和胰岛素测定,计算体重指数(BMI)、腰臀比、胰岛素作用指数(IAI)和稳态模型评估的胰岛素抵抗指数(HOMA-IR),所有受试者均接受正常血糖高胰岛素钳夹试验.结果 IAI、HOMA-IR及腰围仅在NGT组中与葡萄糖清除率(GDR)具有相关性,腰臀比在NGT组和2型糖尿病组中与GDR具有相关性,而BMI则在NGT组、IGT组和2型糖尿病组中均与GDR具有相关性.结论 使用常用的胰岛素抵抗评价方法尤其HOMA-IR要注意其适用条件,避免不适当应用.     

应用高葡萄糖钳夹技术评价女性非酒精性脂肪性肝病胰岛β细胞功能和胰岛素抵抗

目的评价女性非酒精性脂肪性肝病(NAFLD)胰岛β细胞功能和胰岛素抵抗。方法应用高葡萄糖钳夹技术检测了9例NAFLD和7例正常对照组胰岛β细胞功能和胰岛素敏感性,采用梯度回波化学位移MR I评估了肝脏同相(IP)像值和反相(OP)像值,并测定了皮下脂肪面积(SA)、内脏脂肪面积(VA)。结果NAFLD组空腹胰岛素(FINS)、胰岛素分泌第一时相(1Ph)、葡萄糖代谢清除率(GDR)、胰岛素敏感指数(ISI)低于正常对照组(P<0.05)。结论女性NAFLD患者存在早期胰岛功能损害和胰岛素抵抗。     

经典1型糖尿病患者胰岛素抵抗的高胰岛素正葡萄糖钳夹研究

目的 应用高胰岛素正葡萄糖钳夹技术评价1 型糖尿病(T1DM)患者胰岛素抵抗程度.方法 选择新诊断T1DM患者8例,初诊未治2型糖尿病(T2DM)患者8例,均经2周胰岛素强化治疗后进行高胰岛素正葡萄糖钳夹检测胰岛素敏感性指数(ISI),并与正常糖耐量志愿者10例进行比较.结果 T1DM组的年龄、BMI、空腹胰岛素(FIns)及空腹C-肽等各项参数显著低于正常对照组(P＜0.05),而腰臀比(WHR)、收缩压(SBP)、舒张压(DBP)、TC、TG、LDI-C、HDL-C均与正常对照组差异无统计学意义.T1DM组与T2DM组比较,年龄、BMI、WHR、FIns、空腹C-肽、SBP、TC、TG均显著低于T2DM组(P＜0.05).正常对照组、T1DM组、T2DM组ISI分别为12.83±1.09、9.95±0.50、3.80±0.20,3组间ISI差异有统计学意义(P＜0.05).结论 高胰岛素正葡萄糖钳夹检测结果显示,TIDM患者的ISI显著低于正常人,但高于T2DM患者.     

用葡萄糖钳夹技术评价各种简易胰岛素抵抗指数计算公式

目的探讨以金标准正糖钳夹技术评价各种简易胰岛素抵抗指数计算公式的准确性。方法选取糖耐量正常（NGT）者14例，IGT者23例，T2DM者8例，以钳夹技术及各简易胰岛素抵抗指数计算公式（1／Fins，FPG／FIns，HOMA—IR，IAI，QUICKI）测定IR程度，以钳夹技术试验结果葡萄糖输注速率（GIR）为评价标准，对各个指数进行评价。结果QUICKI、IAI、HOMA—IR、FIns在NGT组和IGT组中与GIR显著相关，在T2DM组不相关。与GIR的相关程度以QUICKI、IAI最强、HOMA—IR及1／FIns次之。FPG／FIns与GIR无明显相关。结论在NGT和IGT人群中，QUICKI、IAI、HOMA—IR和1／FIns能够比较准确地评估IR程度；但在Ins分泌严重缺陷的T2DM人群中，应用则受到限制。     

长效胰岛素类似物葡萄糖钳夹研究结果分析

高胰岛素-正葡萄糖钳夹技术通过同时外源性输注胰岛素和葡萄糖,将血糖维持在设定的稳态,最初用于评价机体的胰岛素敏感状况,近年来也用于评估胰岛素制剂的药理学特点,被公认为是评价胰岛素作用的"金标准".     

血糖钳技术评价两种胰岛素抵抗动物模型

目的建立血糖钳方法,检验所建高脂、高果糖饮食诱发的胰岛素抵抗模型以及谷氨酸钠(monosodiumglutamate,MSG)所致的肥胖性大鼠胰岛素抵抗模型.方法腹腔注射小剂量链脲霉素(streptozotocin,STZ),30mg/kg轻度损伤胰岛β细胞,造成大鼠糖耐量异常后加喂高果糖高脂肪饲料,并饮用10%果糖水,造成胰岛素抵抗.吡格列酮20mg/kg@d-1治疗后,以血糖钳方法测定葡萄糖输注速率(glucoseinfusionrate,GIR);另一模型为具胰岛素抵抗特征的MSG肥胖性大鼠.新生Wistar大鼠出生第二天起皮下注射L-谷氨酸钠4g/kg@d1,连续7天,造成肥胖.10月龄时,用中药复方千金黄连丸加味提取物,相当于生药10g/kg@d-1治疗,并以血糖钳方法测定对葡萄糖输注速率的影响,且分析了反映胰岛素敏感性的GIR值同血脂、脂肪重、体重、血胰岛素水平及胰岛素敏感指数等指标的相关性.结果两模型组同相应正常对照组相比,胰岛素敏感性均明显降低,给药后均有显著提高.且GIR反映的敏感性同胰岛素敏感指数呈较好的正相关,而与空腹血胰岛素水平、血甘油三酯、血总胆固醇以及腹脂重、腹脂/体重值呈负相关.结论高脂高果糖饮食诱发的大鼠模型及MSG肥胖性大鼠均具有明显的胰岛素抵抗,分别以吡格列酮以及中药复方提取物治疗后,抵抗性均有明显改善.     

Evaluation of insulin sensitivity in patients with Klinefelter's syndrome: a hyperinsulinemic euglycemic clamp study

Patients with Klinefelter’s syndrome have a higher incidence of diabetes mellitus and the percentage of insulin resistance was reported to be high in these patients. However, little is known about the insulin sensitivity assessed by the hyperinsulinemic euglycemic clamp in these patients. In the present study, subjects included 13 newly diagnosed patients with Klinefelter’s syndrome, and 9 age- and body mass index-matched healthy males. The hyperinsulinemic euglycemic clamp was performed in all patients and controls. Insulin resistance was present in five (38.5%) patients with Klinefelter’s syndrome. Compared with control subjects, patients with Klinefelter’s syndrome had elevated plasma concentrations of fasting insulin, follicle-stimulating hormone, luteinizing hormone, estradiol, and sex hormone-binding globulin, whereas they had reduced plasma free testosterone and total testosterone concentrations. The multivariate linear regression analysis showed that fasting glucose, fasting insulin, free testosterone, and total testosterone were independently associated with M-value. In conclusion, the present study by using hyperinsulinemic euglycemic clamp indicates the high prevalence of insulin resistance in Klinefelter’s syndrome patients. However, these patients did not have reduced mean M-values compared with the controls, although their plasma insulin levels were significantly elevated. It is possible that hyperinsulinemia may be the primary metabolic abnormality rather than insulin resistance.     

小鼠扩展胰岛素钳夹术中糖代谢的变化

目的建立小鼠扩展胰岛素钳夹技术并观察其体内糖代谢的变化。方法分别采用[3-^3H]葡萄糖和2-脱氧-^3H葡萄糖（2-DOG）作为示踪剂,建立自由状态下小鼠正糖-高胰岛素钳夹技术,并观察其体内糖代谢的动态改变。结果在钳夹稳定状态,血糖稳定在基础水平（7.0±1.3mmol/L）,在高血浆胰岛素水平下,血浆FFA明显抑制（从1.45±0.15到0.45±0.08mmol/L,P〈0.01）。葡萄糖输注率为46.7±4.7mg·kg^-1·min^-1,葡萄糖清除率为11.2±0.6mg·kg^-1·min^-1,肝糖产生被完全抑制,骨骼肌葡萄糖摄取率为20.9±2.5μmol·100g^-1·min^-1,而TG和TC浓度较钳夹前明显降低（P均〈0.01）。平均血糖变异系数为5.19%,平均GIR变异系数为9%。结论采用上述示踪剂建立的小鼠扩展胰岛素钳夹技术,能准确评价小鼠机体胰岛素敏感性,同时能动态反映体内糖代谢变化。     

Evaluation of insulin sensitivity in hyperprolactinemic subjects by euglycemic hyperinsulinemic clamp technique

The background and aim of the study is to evaluate insulin sensitivity in hyperprolactinemic subjects via euglycemic hyperinsulinemic clamp technique. Sixteen hyperprolactinemic subjects and 12 healthy subjects were included in the study. HOMA-B and HOMA-IR values of groups were calculated. Euglycemic hyperinsulinemic clamp technique was performed in both groups, and the M value of the groups was defined. Mann–Whitney U and chi-square tests were used in statistical analysis. Basal insulin level of hyperprolactinemic patients were higher than the control group (6.85 ± 4.68; 3.66 ± 0.88 μU/ml respectively; P < 0.05). Mean HOMA-IR and HOMA-B values of patients were higher than control group (1.49 ± 1.30; 0.78 ± 0.27 respectively; P = 0.02 and 136.28 ± 72.53; 64.77 ± 23.31, respectively, P < 0.001). M values of the patients were statistically lower than the control group (5.64 ± 2.36; 7.05 ± 1.62 kg/mg/min respectively; P < 0.05). (1) Hyperprolactinemic patients were more insulin resistant than control subjects. (2) Insulin resistance in hyperprolactinemic patients is not associated with obesity or anthropometric parameters such as fat content, waist circumference and BMI.     

Insulin sensitivity assessment in uncomplicated obese women: comparison of indices from fasting and oral glucose load with euglycemic hyperinsulinemic clamp

BACKGROUND AND AIM: Obesity is associated with a great variability to insulin sensitivity degree. Several formulae developed from measurements in the fasting state and during the oral glucose tolerance test (OGTT) have been proposed to assess insulin sensitivity. AIM: In this work we sought to compare the published insulin sensitivity indices with the metabolized glucose index obtained by hyperinsulinemic euglycemic clamp in uncomplicated obese subjects. Uncomplicated obesity provides a good model in order to study insulin sensitivity per se. METHODS AND RESULTS: In this protocol, 65 obese women affected by uncomplicated obesity (without impaired glucose tolerance, diabetes, hypertension and dyslipidemia) underwent 2 h OGTT and euglycemic hyperinsulinemic clamp. Common formulae obtained in the fasting state and from a 2h OGTT were calculated. Simple linear regression analysis showed that ISI (r=0.592, p=0.01), 2 h OGIS (r=0.576, p=0.02), MCRest (r=0.507, p=0.02), 120 insulin (r=-0.494, p=0.03) and fasting insulin (r=-0.382, p =0.02) are significantly correlated to the M index obtained from the hyperinsulinemic euglycemic clamp. The Bland-Altman plot confirmed the good agreement between indices from OGTT and the clamp. CONCLUSION: OGTT-derived indices provide a good assessment of insulin sensitivity in obesity. OGTT could easily be applied in a large number of obese patients in order to obtain information on both glucose tolerance and insulin sensitivity.