乌司他丁对重度脓毒症诱导的ALI/ARDS模型大鼠肺组织中蛋白C及活化蛋白C含量的影响

目的:探讨乌司他丁对重度脓毒症诱导的急性肺损伤(acute lung injury,ALI)/急性呼吸窘迫征(acute respiratory distress syndrome,ARDS)模型大鼠肺组织中蛋白C(protein C,PC)及活化蛋白C(activated protein C,APC)含量的影响.方法:对27只体质量为200～250 g的清洁级雄性SD大鼠采用盲肠结扎穿孔法联合0.9％氯化钠溶液灌洗法制作重度脓毒症诱导的ALI/ARDS大鼠模型,以PaO2/FiO2≤300作为ALI/ARDS模型大鼠纳入标准,随机分为对照组、乌司他丁5万U/kg组、乌司他丁10万U/kg组,每组9只.各组均在造模成功后立即予乌司他丁或0.9％氯化钠溶液静脉注射,乌司他丁5万U/kg组予乌司他丁5万U/kg静脉注射,乌司他丁10万U/kg组予乌司他丁10万U/kg静脉注射,对照组予0.9％氯化钠溶液静脉注射,静脉注射持续时间为10 min.造模后1、2、3h分批处死大鼠,取肺组织制备组织匀浆,采用酶联免疫吸附试验(ELISA)法测定其PC及APC含量.结果:(1)乌司他丁10万U/kg组大鼠造模后1、2、3h肺组织PC的含量分别为(5196±981)ng/mL、(6156±1256) ng/mL、(6481±1141) ng/mL,均高于乌司他丁5万U/kg组[(3691±812) ng/mL、(4367±1547)ng/mL、(5313±1356)ng/mL]和对照组[(3676±579)ng/mL、(3248±851)ng/mL、(3188±1808) ng/mL],且差异有统计学意义(P＜0.01);(2)乌司他丁10万U/kg组大鼠造模后1、2、3h肺组织APC含量分别为(2798±380)ng/mL、(2964±731)ng/mL、(3156±805)ng/mL,均高于乌司他丁5万U/kg组[(1596±516)ng/mL、(1994±196) ng/mL、(1712±456)ng/mL],但差异无统计学意义(P＞0.05);两组APC含量均高于对照组[(1459±395)ng/mL、(1300±348)ng/mL、(1141±318) ng/mL],差异均有统计学意义(P＜0.05).各组大鼠PC和APC含量组内比较差异均无统计学意义(P＞0.05).结论:乌司他丁有助于提高重度脓毒症诱导的ALI/ARDS模型大鼠肺组织PC和APC的含量,并呈剂量依赖趋势,它可能对脓毒症诱导的ALI/ARDS大鼠肺组织有保护作用.     

饱和氢气生理盐水对油酸致急性肺损伤大鼠肺的保护作用

目的 探讨饱和氢气生理盐水是否对油酸所致大鼠急性肺损伤具有保护作用.方法 将健康成年SD大鼠30只,随机分为对照组、油酸+生理盐水组和油酸+饱和氢气生理盐水组,每组10只.对照组(假手术组):予股动脉置管留取血气标本,待血流动力学稳定后经尾静脉注入生理盐水0.1 mL/kg;油酸+生理盐水组:注入高纯度油酸0.1 mL/kg,余同对照组;油酸+饱和氢气生理盐水组:注入高纯度油酸5 min后给予腹腔注射饱和氢气生理盐水5 mL/kg,余同油酸+生理盐水组;同时予前两组腹腔注射等量生理盐水,3 h后再次留取动脉血标本并处死大鼠.检测血气分析,肺组织髓过氧化物酶(MPO)活性、丙二醛(MDA)及肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和NF-κB p65水平;观察肺组织病理变化.结果 与注射油酸前比较,生理盐水治疗组和饱和氢气生理盐水治疗组在注射油酸3 h后PaO2显著降低(P<0.05),且显著低于对照组(P<0.05).此外,饱和氢气生理盐水治疗组PaO2显著高于生理盐水治疗组(P<0.05).与对照组比较,生理盐水治疗组和饱和生理氢气盐水治疗组肺组织MPO活性及MDA水平显著增加(P<0.01,P<0.05),且饱和氢气生理盐水治疗组显著低于生理盐水治疗组(P<0.05).生理盐水治疗组和饱和氢气生理盐水治疗组肺组织TNF-α、IL-1β和NF-κB p65水平显著高于对照组(P<0.05),且饱和氢气生理盐水治疗组显著低于生理盐水治疗组(P<0.05).结论 饱和氢气生理盐水能够降低大鼠油酸肺损伤模型中肺的损伤程度,其机制可能与氢分子在体内的选择性抗氧化作用有关.     

胰岛素与硒联合作用糖尿病大鼠骨骼肌胰岛素信号转导通路中PI3K和GLUT4蛋白的表达

背景:PI3K是骨骼肌中胰岛素信号转导途径级联反应中关键的蛋白分子之一,其表达异常可影响GLUT4的合成、分泌、移位等变化,从而使血糖升高.目的:通过检测胰岛素和硒联合应用对糖尿病大鼠骨骼肌胰岛素信号转导通路有关蛋白激酶(PI3K、GLUT4)表达的影响.方法:SD雄性大鼠随机分为正常组、糖尿病组、糖尿病+胰岛素组、糖尿病+亚硒酸钠组、糖尿病+胰岛素+亚硒酸钠组.除正常组外,其余4组大鼠腹腔注射链尿佐菌素50 mg/kg复制大鼠糖尿病模型.正常组和糖尿病自由进水和进食;糖尿病+胰岛索组按1 U/(kg·d)皮下注射胰岛素,糖尿病+亚硒酸钠组按180 μg/(kg·d)管饲亚硒酸钠,糖尿病+胰岛素+亚硒酸钠组两药联合用药持续4周;应用免疫印迹和免疫组织化学法检测各实验组骨骼肌胞浆中PI3K和胞膜上GLUT4蛋白表达.结果与结论:免疫组织化学方法所得结果与免疫印迹一致.胰岛索和硒联合应用能明显增加骨骼肌细胞胞浆中PI3K和胞膜上GLUT4蛋白的表达量,说明胰岛索和硒联合应用是通过PI3K途径和增加骨骼肌组织GLUT4蛋白的表达来增强胰岛素信号转导.     

二甲双胍对糖尿病大鼠心肌保护作用的形态学观察

目的:观察传统的降糖药二甲双胍对糖尿病大鼠心肌的保护作用。方法:实验于2004-03/10在锦州医学院试验动物中心完成。①取雄性SD大鼠一次性尾静脉注射新鲜配置的20g/L链脲佐菌素溶液60mg/kg制备糖尿病模型,取造模成功(空腹血糖≥13.9mmol/L)大鼠50只单纯随机分为糖尿病组10只,胰岛素组和二甲双胍组各20只,另取10只尾静脉注射枸橼酸钠缓冲液的正常大鼠为正常对照组。②成模后第4天起,糖尿病组隔天皮下注射鱼精蛋白胰岛素1～2U/次维持存活,胰岛素组皮下注射普通胰岛素(4～6U/次,每天两三次),二甲双胍组灌胃二甲双胍20～30mg/(kg·d),正常对照组每天灌胃等量生理盐水。实验过程中胰岛素组和二甲双胍组大鼠空腹血糖控制在6.4mmol/L以下。③治疗12周后,大鼠断头处死,采血测血糖和血浆胰岛素,取左心室心肌组织,常规制作电镜切片,JEOL1200EX电子显微镜观察。结果:60只大鼠进入结果分析。①空腹血糖水平:胰岛素组和二甲双胍组显著低于糖尿病组[(7.84±3.6),(8.16±3.13),(14.00±4.00)mmol/L,P<0.05],与正常对照组比较差异不显著[(5.76±0.84)mmol/L,P>0.05]。②空腹血浆胰岛素水平:二甲双胍组与正常对照组比较差异不明显[(11.01±4.21),(9.48±3.56)mU/L,P>0.05],但低于胰岛素组和糖尿病组[(33.86±14.73),(23.39±13.85)mU/L,P<0.05]。③心肌超微结构:糖尿病组大鼠心肌纤维内肌节紊乱,肌原纤维变性,线粒体增多,常有肿胀变性。经二甲双胍和胰岛素治疗后糖尿病大鼠心肌超微结构病理改变明显减轻。结论:二甲双胍能抑制糖尿病大鼠的心肌病变,对心肌有良好的保护作用。     

槟榔碱对2型糖尿病大鼠胰腺β细胞PDX-1 mRNA表达的影响

目的:探讨槟榔碱对2型糖尿病大鼠β细胞分泌功能及胰腺十二指肠同源盒因子-1(pancreas-duodenum homeobox-1,PDX-1)mRNA表达的影响。方法:采用高果糖饲料饲养Wistar大鼠12周制备2型糖尿病大鼠模型,40只实验动物随机分为5组:对照组、模型组、模型+1 mg/kg槟榔碱组、模型+5 mg/kg槟榔碱组、模型+10 mg/kg槟榔碱组。药物注射4周后股动脉取血检测空腹血糖、三酰甘油、总胆固醇、高密度脂蛋白、低密度脂蛋白、血清胰岛素,处死大鼠取胰腺组织,石蜡切片HE染色观察组织形态学变化,RT-PCR检测β细胞内PDX-1及胰岛素mRNA的表达水平。结果:(1)高糖作用引起Wistar大鼠胰腺β细胞PDX-1及胰岛素mRNA的表达水平显著下降(P0.01);(2)1 mg/kg和5 mg/kg槟榔碱处理能显著上调高糖喂养Wistar大鼠胰腺β细胞PDX-1与胰岛素mRNA表达水平(P0.05,P0.01)。结论:槟榔碱可以通过上调PDX-1和胰岛素基因表达,改善高糖环境下的β细胞胰岛素合成和分泌功能的损伤。     

一种基于硫酸葡聚糖铁复合物的胰岛素缓释微胶囊及其降血糖活性

背景：利用各种不同聚合物为载体材料，包裹胰岛素等蛋白多肽类药物的微球缓释系统有可能克服此类药物稳定性差、体内半衰期短、易水解变性的缺陷。目的：制备一种胰岛素缓释微胶囊，并观察其体外释放效果和体内降血糖活性。设计、时间及地点：对比观察实验，于2007—05／2008-01在哈尔滨工业大学生物医学中心纳米药物与生物传感器实验室完成。材料：以硫酸葡聚糖和Fe^3＋为壁材，采用静电吸引层层自组装技术制备胰岛素缓释微胶囊INS（DS／Fe^3＋）。Wistar雄性大鼠腹腔注射链脲佐菌素制备糖尿病大鼠模型。方法：通过体外释放实验观察INS（DS／Fe^3＋）的缓释效果。取糖尿病模型大鼠18只，随机分成3组，其中两组分别皮下注射胰岛素注射液5U／kg或胰岛素微胶囊100U／kg，第3组灌胃胰岛素微胶囊100U／kg。主要观察指标：胰岛素微胶囊的包封率、载药量、体外释放效果及体内降血糖活性。结果：胰岛素缓释微胶囊INS（DS／Fe^3＋）的包封率和载药量分别在60％和40％以上：体外释放实验显示INS（DS／Fe^3＋）有较好的缓慢释放特性，随着包裹层数增加，药物释放速度减慢；体内活性实验表明皮下注射胰岛素微胶囊在体内能够保持6～10h的降血糖效果，灌胃给药未表现出降血糖效果。结论：以硫酸葡聚糖铁为载体，静电吸引层层自组装法制备的胰岛素缓释微胶囊稳定性好，能够维持较长时间的降血糖效果。     

胰岛素的气雾疗法

或许有一天你同病人讨论如何进行胰岛素吸入治疗 ,药物经肺粘膜吸收而进入体循环。目前气雾吸入胰岛素仍处于研究早期。如果研究论证了这种给药模式 ,那么对于因注射胰岛素而给生活带来不便的人群来说无疑是一大惊喜。7名患 型糖尿病的男女患者 (全部口服降糖药 )参与了一项研究。他们按 1 .5 U/kg的标准在餐前 5 min气雾吸入胰岛素监测血糖及胰岛素水平。在另一时段 ,同样的自愿者吸入安慰气雾剂以作对照。而在这两次试验中他们并不清楚自己吸入了什么。该 7名受试者在停用降糖药 4d后改用胰岛素喷雾剂的早晨 ,按 1 U/kg剂量吸入胰岛素 (…     

硫化氢对油酸诱导急性肺损伤大鼠细胞外基质的调节作用

目的 探讨外源性硫化氢(H2S)对油酸诱导的急性肺损伤(ALI)大鼠细胞外基质(ECM)的调节作用.方法 雄性SD大鼠35只,随机分为对照组(n=5)、油酸组(n=15)、油酸+硫氢化钠(NaHS)组(n=15).油酸组大鼠尾静脉注射油酸0.1 mL/kg;油酸+NaHS组先腹腔注射NaHS 56 μmol/kg(溶于0.5 mL生理盐水中),30 min后再尾静脉注射油酸0.1 mL/kg.以上两组均分为2、4、6 h三个观察点,每个点5只.对照组大鼠尾静脉注射0.1 mL/kg生理盐水,观察6 h.在观察时间结束后,麻醉大鼠,行支气管肺泡灌洗,灌洗液(BALF)沉渣行瑞士染色进行中性粒细胞(PMN)分类计数;取右肺上叶,计算湿重/干重(W/D)值;用ELISA法检测血浆、肺组织匀浆基质金属蛋白酶-9(MMP-9)、组织金属蛋白酶抑制剂-1(TIMP-1)水平;用敏感硫电极法检测H2S水平.结果 与对照组比较,油酸组2、4、6 h PMN显著升高(P均<0.01);与油酸组比较,油酸+NaHS组各时间点PMN显著降低(P均<0.05).与对照组比较,油酸组2、4、6 h肺W/D值显著升高(P均<0.01);与油酸组比较,油酸+NaHS组4、6 h肺W/D值显著降低(P均<0.05).与对照组比较,油酸组2、4、6 h血浆、肺组织匀浆H2S水平显著降低(P均<0.01);与油酸组比较,油酸+NaHS组血浆、肺组织匀浆H2S水平4、6 h显著升高(P均<0.05).与对照组比较,油酸组2、4 h血浆、肺组织匀浆及6 h血浆MMP-9水平显著升高(P均<0.05);与油酸组比较,油酸+NaHS组4 h 血浆MMP-9水平显著降低(P<0.05),2、6 h血浆、肺组织匀浆及 4 h 肺组织匀浆MMP-9水平降低,但差异无统计学意义(P均>0.05).与对照组比较,油酸组4 h 血浆、肺组织匀浆及6 h肺组织匀浆TIMP-1水平显著降低(P均<0.05);与油酸组比较,油酸+NaHS组2、4、6 h血浆、肺组织匀浆TIMP-1水平升高,但差异无统计学意义(P均>0.05).结论 外源性H2S可能通过调节肺组织MMP-9和TIMP-1水平的变化,发挥对油酸诱导大鼠ALI 的细胞外基质的调节作用.     

夏枯草及抗炎1号注射液经肝动脉联合灌注 治疗大鼠晚期肝癌的实验研究

目的:观察夏枯草及抗炎1号注射液经肝动脉联合灌注对大鼠晚期肝癌的治疗作用。方法:建立30只二乙基亚硝胺(DEN)诱发大鼠晚期原发性肝癌模型,随机分为中药组、化疗组和盐水组,每组各10只。经胃、十二指肠动脉至肝固有动脉分别灌注生理盐水0.5mL、夏枯草注射液0.6g/kg+抗炎1号注射液0.45g/kg、顺铂(DDP)4mg/kg+羟基喜树碱(HCPT)1.5mg/kg+5-氟脲嘧啶(5-FU)34mg/kg。结果:与化疗组和盐水组比较,中药组大鼠的一般情况较好,肝重/体重低,病理性肝损害轻,生存率高(90％对60％,70％,P〈0.05)。结论:经肝动脉联合灌注夏枯草及抗炎1号注射液较灌注化疗药物对诱发性大鼠晚期肝癌具有更好的作用。     

糖尿病早期大鼠坐骨神经电生理改变及胰岛素的影响

目的:观察糖尿病早期大鼠坐骨神经电生理改变及胰岛素对其的影响。方法:实验于2005-09在南通大学药理教研室完成。从18周龄健康雄性SD大鼠36只中随机数字表法取28只造模,8只作为正常对照组。给予大鼠一次性腹腔注射链脲菌素造成胰岛素依赖性糖尿病模型,正常对照组给予等体积的柠檬酸盐缓冲液。给药72h后造模大鼠尾静脉取血,邻甲苯胺法测定血糖浓度,非空腹血糖大于16.0mmol/L纳入糖尿病模型。将纳入糖尿病模型的大鼠随机分为糖尿病模型组和胰岛素治疗组。造模成功后的第2天,每天给予胰岛素治疗组大鼠皮下注射精蛋白锌胰岛素注射液(长效胰岛素,40U/mL),给药时间在下午5点左右,胰岛素起初剂量4U/只,次日上午测定其血糖值,控制大鼠非空腹血糖值在10mmol/L以下(不低于3.9mmol/L),调整胰岛素用量为4～6U。2周时停止给药,并再次测定大鼠的血糖和体质量。糖尿病模型组不作任何处理。糖尿病模型组与正常对照组与胰岛素治疗组同时间进行血糖和体质量的测定。麻醉各组大鼠分离、暴露坐骨神经,测定其感觉神经动作电位潜伏期、波幅及传导速度。结果数据行方差分析,两两比较使用Scheffe检验。结果:在28只造模SD大鼠给予链脲菌素后第72h,有15只大鼠血糖值为16.05～20.89mmol/L,超过16.0mmol/L,造模成功,纳入糖尿病模型。将造模成功的糖尿病模型大鼠分为糖尿病模型组8只,胰岛素治疗组7只;和正常对照组大鼠8只一起进入结果分析。①糖尿病模型组坐骨神经感觉神经动作电位的传导速度与波幅和正常对照组相比显著降低,差异有显著性意义[(40.34±1.68),(55.47±9.05)m/s,P<0.01]O[(510.73±22.10),(637.37±29.28)μV,P<0.01]O潜伏期延长[(1.54±0.13),(1.19±0.13)ms,P<0.01]。给予胰岛素治疗后能有效逆转这些变化,坐骨神经感觉神经动作电位传导速度增加至(48.57±1.86)m/s,波幅上升至(593.72±22.62)μV,潜伏期缩短至(1.31±0.06)ms,与糖尿病模型组比较差异有显著性意义。②造模后2周糖尿病模型组血糖值显著高于正常对照组,差异有显著性意义[(20.1±2.0),(5.1±0.6)mmol/L,P<0.01];给予胰岛素治疗后血糖值显著降低至(6.6±1.8)mmol/L,与糖尿病模型组比较差异有显著性意义(P<0.01)。结论:在糖尿病早期(2周)即存在感觉神经功能的损害,起始因素为高血糖,通过胰岛素控制血糖后,能有效防止神经病损。     

Intra-individual variability of the metabolic effect of inhaled insulin together with an absorption enhancer

OBJECTIVE: To study the metabolic effect and the variability of the effect elicited by inhalation of 87.2 U insulin powder combined with an absorption enhancer. The metabolic effect was compared with that of 10.2 U regular insulin injected subcutaneously and of 5.5 U regular insulin given intravenously RESEARCH DESIGN AND METHODS: In this single-center open euglycemic glucose clamp study 13 healthy male volunteers received 5 insulin administrations on separate study days: once as an intravenous dose, once as a subcutaneous injection, and 3 times by inhalation, in randomized order. Glucose infusion rates (GIRs) necessary to keep blood glucose concentrations constant at 5.0 mmol/l were determined over an 8-h period after administration. RESULTS: After inhalation of the insulin powder aerosol, the onset of action was substantially more rapid than after subcutaneous insulin injection, and maximal action was reached earlier (86+/-47 vs. 182+/-53 min, P<0.0001). The maximal glucose infusion rate after inhalation of insulin was comparable to that after subcutaneous insulin injection (9.2+/-2.6 vs. 8.8+/-2.8 mg x kg(-1) x min(-1), NS). The metabolic effect in the first 2 h after inhalation was significantly greater than that after subcutaneous insulin injection (amount of glucose infused: 0.88+/-0.25 vs. 0.59+/-0.20 g x kg(-1) x 120 min(-1), P<0.0001). However, the total metabolic effect after inhalation and subcutaneous injection was comparable (2.50+/-0.76 vs. 2.56+/-0.69 g x kg(-1) x 480 min(-1), NS). The relative bioefficacy of inhaled insulin calculated in relation to the data from the subcutaneous insulin application was 12.0+/-3.5% (absolute bioefficacy 10.1+/-3.1%) but was highest in the first 2 h after application (18.5+/-3.7%; absolute bioefficacy 8.2+/-4.1%). The intraindividual variability of the metabolic response induced by insulin inhalation was 14+/-9% for the maximal glucose infusion rate, 15+/-10% for the time-to-maximal effect, and 16+/-12% for the total amount of glucose infused. CONCLUSIONS: This feasibility study shows that inhaled insulin with an absorption enhancer has a pronounced metabolic effect compared with the results of a previous study of inhaled insulin without an enhancer. The intraindividual variability of the metabolic effect was comparable with that of inhaled and subcutaneously injected insulin.     

低血糖对肌肉损伤影响的实验研究

目的;研究胰岛素剂量和低血糖持续时间对血清酶活性影响,确定低血糖损伤的脏器。方法：根据胰岛素剂量和低血糖持续时间的不同,将30只家兔分为5组：A2组,胰岛素2U／kg低血糖持续30min;A10组,胰岛素10U/kg低血糖持续30min;B2组,胰岛素2U／kg低血糖持续60min;B10组：胰岛素10U/kg低血糖持续60min;C对照组：胰岛素10U/kg加50％葡萄糖注射不诱发低血糖。胰岛素注射前后对血清酶的活性及肌酸激酶（CK）同功酶的分布进行观察。结果：所有低血糖组其血清CK的活性较对照组明显增高,且在A10和B10组CK活性的升高持续24h,而血清ALT、AST、LDH的升高仅见于B10组。此外,主要存在于心肌和骨骼肌中的CK－Band4在B10组可见显著升高。结论：低血糖所致血清酶活性及CK－Band4的升高是由于肌肉损伤的结果而非肝脏的损伤,低血糖持续的时间和胰岛素剂量可以影响脏器损伤的程度。     

胰岛素治疗高血压动脉硬化性脑梗死的实验研究

目的：观察胰岛素（ Ｉｎｓ）对高血压动脉硬化大鼠脑梗死的疗效。方法：５０ 只肾血管性高血压大鼠（ Ｒ Ｈ Ｒ）复制成大脑中动脉闭塞（ Ｍ Ｃ Ａｏ）模型,随机分４ 组： Ａ 组１２ 只（ Ｉｎｓ ２１ Ｕ／ｋｇ）, Ｂ组１２ 只〔 Ｉｎｓ ２１ Ｕ／ｋｇ＋ ５０％ 葡萄糖（２ ｇ／ｋｇ）〕, Ｃ组 １２ 只〔 Ｉｎｓ ４５ Ｕ／ｋｇ＋ ５０％ 葡萄糖（４ ｇ／ｋｇ）〕和 Ｄ组 １４ 只（生理盐水 ４ ｍ ｌ／ｋｇ）。各组均于 Ｍ Ｃ Ａｏ 后即注射胰岛素, Ｍ Ｃ Ａｏ 后 ４ 小时和２４ 小时检查神经功能,２４ 小时处死大鼠取脑,测大脑体积和梗死灶体积。结果： Ａ 组的血糖较其他组有统计学意义的下降（ Ｐ均＜ ００１）, Ｃ组的神经功能障碍评级、梗死灶体积及其与大脑体积的百分比的减少都有统计学意义（ Ｐ 均＜ ００１）, Ａ、 Ｂ、 Ｄ组间比较则无差异（ Ｐ 均＞００５）。结论：胰岛素对缺血脑组织具有不依赖于其降糖作用的直接保护作用, Ｒ Ｈ Ｒ Ｍ Ｃ Ａｏ 后注射胰岛素在较高剂量时才显示疗效,这可能与高血压致脑血管发生病变有关。     

Cotherapy with recombinant human insulin-like growth factor I and insulin improves glycemic control in type 1 diabetes. RhIGF-I in IDDM Study Group

OBJECTIVE: To study the effects of 12 weeks of cotherapy with recombinant human IGF-I (rhIGF-I) and insulin on glycemic control in patients with type 1 diabetes. RESEARCH DESIGN AND METHODS: The study population consisted of 223 patients who ranged in age from 11-66 years and were randomized in a double-blind study to receive 12 weeks of treatment with twice-daily subcutaneous injections of placebo (n = 54), or rhIGF-I at a dose (A.M/P.M) of 40/40 micrograms/kg (n = 56), 80/40 micrograms/kg (n = 57), or 80/60 micrograms/kg (n = 56), while continuing to receive standard insulin therapy. Patients were instructed to test blood glucose levels four times daily and adjust insulin doses to optimize blood glucose control. HbAlc, insulin requirements, body weight, and parameters of the IGF-IGF-binding protein axis were assessed before and during treatment. RESULTS: All groups were comparable at baseline with respect to mean age, gender distribution, duration of diabetes, HbAlc, and BMI. Cotherapy with rhIGF-I/insulin produced a mean decrease in HbAlc of 1.2%, compared with a 0.7% decrease in HbAlc for patients receiving intensified insulin therapy alone (P < or = 0.01). Subjects receiving rhIGF-I/insulin cotherapy also decreased their daily insulin usage by 11-19%, compared with a 7% increase in insulin usage reported by the placebo group. Moreover, the incidence of hypoglycemia was similar in subjects treated with rhIGF-I/Insulin cotherapy compared with those treated with insulin alone, despite the better glycemic control of the former group. The 40/40 dose of rhIGF-I was well tolerated. Higher doses of rhIGF-I did not further improve efficacy yet were associated with unacceptable levels of adverse events, including edema, jaw pain, and early worsening of retinopathy. CONCLUSIONS: These results demonstrate that rhIGF/insulin cotherapy improves glycemic control in patients with type 1 diabetes better than optimized insulin management alone; longer-term trials would be required to determine an acceptable benefit-risk profile.     

Control of postprandial plasma glucose by an oral insulin product (HIM2) in patients with type 2 diabetes

OBJECTIVE: The objectives of this exploratory study were to assess the postprandial glucose-lowering effects and evaluate the safety and tolerability of single, escalating doses of an oral insulin product, hexyl-insulin monoconjugate 2 (HIM2), in patients with type 2 diabetes. Subcutaneous insulin and oral placebo were also administered for comparison. RESEARCH DESIGN AND METHODS: Eighteen patients with type 2 diabetes were enrolled in this randomized, single-blind, placebo-controlled, three-way crossover, dose-escalation study. A single dose of each of the following study drugs was administered to each patient on 3 separate days: oral HIM2 (at one of three dose levels: 0.375, 0.5, or 1.0 mg/kg), subcutaneous regular insulin (8 units Humulin R), and oral placebo. At 30 min after dosing, patients ingested a standardized test meal (16 oz/720 calories of Boost Plus). Serial blood samples were collected for determination of plasma glucose and insulin concentrations during the 4-h postdose period. RESULTS: The mean glucose area under the curve for 0 to 240 min (AUC(0-240)) values were lower following administration of 0.5 and 1.0 mg/kg HIM2 vs. placebo (1,097.1 vs. 1,196.9 and 801.1 vs. 992.1 mg x h(-1) x dl(-1), respectively). This difference was statistically significant at the 1.0-mg/kg HIM2 dose level. Insulin exposure, as measured by insulin AUC(0-240) values, for the 0.375-, 0.5-, and 1.0-mg/kg dose levels of HIM2 were 169.9, 193.1, and 230.8 micro U x h(-1) x ml(-1), respectively; insulin AUC(0-240) values for placebo were 165.8, 196.1, and 169.2 micro U x h(-1) x ml(-1), respectively. The mean glucose AUC(0-240) values were similar following administration of 0.5 and 1.0 mg/kg HIM2 vs. subcutaneous insulin (1,097.1 vs. 1,048.0 and 801.1 vs. 875.2 mg x h(-1) x dl(-1), respectively). For pooled data from the 0.5- and 1.0-mg/kg dose groups, the HIM2/subcutaneous insulin ratios for the 2-h postprandial glucose concentration (0.97, 95% CI 0.90-1.06), maximum postprandial glucose concentration (0.99, 95% CI 0.93-1.06), and glucose AUC(0-240) (0.98, 95% CI 0.9-1.06) were within 10% of unity, implying glucodynamic equivalence. Although HIM2 (0.5 and 1.0 mg/kg) and subcutaneous insulin (8 units) provided comparable control of postprandial plasma glucose concentrations, HIM2 resulted in peripheral insulin concentrations that were lower than subcutaneous insulin (mean insulin AUC(0-240) of 193.1 vs. 233.6 and 230.8 vs. 270.3 micro U x h(-1) x ml(-1), respectively). CONCLUSIONS: Single, oral doses of HIM2 were safe and well tolerated. HIM2 (0.5 and 1.0 mg/kg) was more effective than placebo and as effective as subcutaneous regular insulin (8 units) at controlling postprandial glycemia with respect to the following parameters: 2-h postprandial glucose concentration, maximum glucose concentration, and glucose AUC(0-240). This occurred even though peripheral insulin concentrations were lower following the administration of HIM2 (0.5 and 1.0 mg/kg) than subcutaneous insulin. Thus, HIM2 therapy may control postprandial glycemia without causing peripheral hyperinsulinemia in patients with type 2 diabetes.     

Combined metformin and insulin therapy for patients with type 2 diabetes mellitus

OBJECTIVE: This study was undertaken to assess the effects of combined treatment with insulin and metformin in patients with type 2 diabetes mellitus in whom dietary measures, weight control, and oral antihyperglycemic therapy had failed. BACKGROUND: Insulin resistance in peripheral tissues, increased hepatic gluconeogenesis, and impaired insulin secretion are the underlying factors in the development of type 2 diabetes. Metformin is a biguanide antihyperglycemic agent that increases peripheral insulin sensitivity, reduces hepatic gluconeogenesis, and decreases intestinal glucose absorption. METHODS: Thirty-one patients (24 women, 7 men; mean age, 61.8 years; mean body mass index [BMI], 28.0 kg/m2) were enrolled in this randomized, double-blind, 2-way, crossover, placebo-controlled study. Patients with type 2 diabetes who were treated previously with insulin or oral hypoglycemic agents and who had a glycosylated hemoglobin (HbA1c) level >9% or a fasting blood glucose level >8 mmol/L were included. Patients who were being treated with oral agents were switched to insulin therapy and required to maintain stable blood glucose control for 2 months prior to randomization. Patients received insulin plus either metformin 1,700 mg/d or placebo for 5 months, followed by a 2-month washout period, and were then crossed over to the other treatment arm for 5 months of additional treatment (total treatment period: 12 months). RESULTS: Thirty patients completed the study; 1 patient withdrew early because of hypoglycemia. Compared with placebo, metformin produced significant reductions from overall baseline in mean daily insulin dose requirement (-8.69 units (17.2%], P < 0.001), HbA1c level (-0.74 [9.9%], P = 0.005), serum fructosamine level (-44.40 micromol/L, P = 0.026), 24-hour blood glucose profile (P = 0.008), and total cholesterol level (-0.42 mmol/L, P = 0.005). No treatment effects were observed on body weight, blood pressure, serum high-density lipoprotein cholesterol levels, or serum triglyceride levels. There was no correlation between BMI and reduction in HbA1C. No major side effects were reported. CONCLUSIONS: Combination therapy with metformin and insulin improves glycemic control and reduces insulin requirements. with no major side effects, in patients with type 2 diabetes and may improve the risk profile in this patient population.     

CordyMax Cs-4 improves glucose metabolism and increases insulin sensitivity in normal rats

OBJECTIVE: To evaluate effects of CordyMax trade mark Cs-4, a mycelial fermentation product of Cordyceps sinensis, on improving glucose metabolism and insulin sensitivity. DESIGN: An in vivo pharmacology study. Subjects and Study Interventions: Adult Wistar rats, male and female, were given CordyMax 250 or 500 mg/kg per day or placebo for 17 days by gavage. OUTCOME MEASUREMENTS: Fasting blood glucose, fasting plasma insulin, glucose-insulin index, and oral glucose tolerance. RESULTS: Rats fed Cs-4 at either 250 or 500 mg/kg showed significantly reduced fasting blood glucose after the 17-day treatment, by 27% and 24% from baselines respectively (both p < 0.001). Examination of fasting plasma insulin demonstrated a 37% decrease in the high dose treatment groups (p = 0.012). Glucose-insulin index, an index of insulin sensitivity, increased by 10% and 17% in both 250 and 500 mg/kg groups (p = 0.008 and p = 0.0001, respectively). Oral glucose tolerance tests showed significantly improved glucose tolerance at 0.5, 1.0, and 2.0 hours after oral administration of a bolus of glucose (the area under the glucose curve: p = 0.05-0.006), but no change at 5 hours. CONCLUSION: CordyMax Cs-4 is effective in lowering basal blood glucose and plasma insulin, improving glucose metabolism by enhancing insulin sensitivity, and improving oral glucose tolerance.     

Lispro Mix25 insulin as premeal therapy in type 2 diabetic patients

OBJECTIVE: Insulin Mix25 is a new premixed insulin analog containing 25% insulin lispro and 75% neutral protamine lispro (NPL) suspension (NPL insulin). The aim of the study was to compare serum glucose and insulin responses after breakfast in type 2 diabetic patients who received Mix25, premixed regular/NPH (30%/70%), or NPH insulin before the meal. RESEARCH DESIGN AND METHODS: We studied 22 type 2 diabetic patients of age 62 +/- 1 years, BMI 30 +/- 1 kg/m2, duration of diabetes 15 +/- 2 years, duration of insulin therapy 6 +/- 1 years, insulin dose 65 +/- 6 U/day, and HbA1c 7.9 +/- 0.2%. Ten healthy individuals (age 56 +/- 1 years, BMI 28 +/- 1 kg/m2) served as control subjects. Each patient (except healthy subjects, who were studied once each) was studied three times in a double-blind, randomized fashion. After an overnight fast, the patients received 36 +/- 4 U of test insulin. Ten minutes after insulin injection, the patients ingested a breakfast meal (512 kcal, 60% carbohydrate, 20% fat, and 20% protein), identical in all studies. Blood samples were taken before and at 10- to 30-min intervals for 240 min after the breakfast meal. RESULTS: The peak rise in serum glucose was lower after Mix25 (76 +/- 7 mg/dl) than after 30/70 (94 +/- 5 mg/dl, P < 0.05) or NPH (113 +/- 4 mg/dl, P < 0.005) insulin. The incremental area under the serum glucose curve was 36% smaller after Mix25 than after 30/70 (P < 0.01) and 56% smaller than after NPH (P < 0.005) insulin. The peak rise in serum insulin concentration was higher after Mix25 (103 +/- 18 mU/l) than after 30/70 (87 +/- 13 mU/l, P < 0.05) or NPH (62 +/- 12 mU/l, P < 0.01) insulin. The incremental area under the serum insulin curve was higher after Mix25 than after 30/70 during the first 2-3 h (P < 0.02), but the difference disappeared by the end of the 4-h follow-up period. After Mix25 injection, there was an inverse correlation between the glucose response to a meal and insulin dose (r = -0.56, P < 0.01) or the incremental area under the serum insulin curve (r = -0.39, P < 0.05). No such correlations were observed with the other insulins. CONCLUSIONS: Because of its faster initial absorption rate, the new premixed insulin analog Mix25 reduces blood glucose response to a breakfast meal in type 2 diabetic patients compared with premixed 30/70 (regular/NPH) or NPH insulin.     

A direct efficacy and safety comparison of insulin aspart, human soluble insulin, and human premix insulin (70/30) in patients with type 2 diabetes

OBJECTIVE: Because there are limited data on the comparison of insulin aspart and mixed insulin in type 2 diabetes, this trial was performed to compare the efficacy and safety of preprandial insulin aspart with human soluble insulin (HI) and human premix (70% NPH/30% regular) insulin (MIX). RESEARCH DESIGN AND METHODS: A total of 231 type 2 diabetic patients were randomized to insulin aspart (n = 75), HI (n = 80), or MIX (n = 76) for 3 months. Insulin aspart and HI were administered with or without bedtime NPH insulin. A total of 204 patients completed the trial according to protocol. HbA1c, 7-point blood glucose, insulin dosage, and hypoglycemic episodes were recorded. The primary end point was "change of HbA1c" from baseline to last visit. Analysis for equivalence was performed by t tests with three subtests. RESULTS: HbA1c decreased 0.91 +/- 1.00 for insulin aspart, 0.73 +/- 0.87 for HI, and 0.65 +/- 1.10 for MIX with the following confidence intervals: insulin aspart HI (-0.21 to 0.57, P = 0.025), insulin aspart MIX (-0.17 to 0.69, P = 0.092), and HI-MIX (-0.33 to 0.48, P = 0.006). Postprandial blood glucose decreased in the insulin aspart group: 0.44 mmol/l to >1.67 mmol/l compared with HI and 1.1 mmol/l to >1.67 mmol/l compared with MIX. Preprandial insulin doses were similar in the insulin aspart and HI groups (10-14.5 U). Hypoglycemic events per month were 0.56 HI, 0.40 insulin aspart, and 0.19 MIX. CONCLUSIONS: Statistically, insulin aspart was not equivalent to another treatment in terms of HbA1c reduction. Insulin aspart treatment resulted in improved HbA1c and postprandial blood glucose. The application of insulin aspart was safe and well tolerated.     

Intravenous insulin infusion to simulate subcutaneous absorption. Bioavailability and metabolic sequelae.

OBJECTIVE: To determine the bioavailability and bioactivity of subcutaneously injected insulin. RESEARCH DESIGN AND METHODS: A randomized block design with six male mongrel dogs as subjects. In protocol 1, purified pork insulin was infused intravenously to simulate the pattern of appearance in the blood that would have been expected from subcutaneous injection. Three intravenous doses (0.05, 0.10, and 0.15 U/kg) were infused on separate days in a pattern (0-300 min) designed to approximately simulate the absorption rate of subcutaneously injected insulin. In protocol 2, interscapular subcutaneous injections of pork insulin at 0.10 U/kg were made. RESULTS: Integrated insulin, decrement in plasma glucose, and maximal glucose clearance for subcutaneous injection experiments were similar to intravenous infusion of equal dose (P greater than 0.10) but significantly different from low-dose infusions (P less than 0.025). Similar results were observed for hepatic glucose output and glucose uptake. Hypoglycemia elicited counterregulatory responses that appeared to be under a threshold differentiated at a plasma glucose of approximately 3 mM. Integrated insulin was plotted against insulin dose to create dose-response curves for intravenous data. The curve was then used to predict the actual appearance rate of insulin in plasma for subcutaneous injection. The estimated bioavailability of subcutaneous insulin was 103.0 +/- 10.5% of the injected dose. CONCLUSIONS: We concluded that, in dogs, insulin delivered subcutaneously in the interscapular area is not significantly degraded before absorption, resulting in metabolic effects equal to intravenous insulin infusion of equivalent dose.