AMPK调节骨骼肌细胞GLUT4基因表达的机制研究

腺苷酸活化蛋白激酶(AMPK)能调节运动/肌肉收缩所引起的骨骼肌细胞葡萄糖转运蛋白4(GLUT4)基因的表达,但至今它的调节机制不清.研究显示在非运动刺激引起的细胞信号事件中由组蛋白去乙酰化酶(HDACs)以及组蛋白乙酰化酶(HATs)控制的组蛋白乙酰化状态是调节基因表达的重要机制,所以我们假设AMPK信号途径是通过征用HDACs中的HDAC5(在骨骼肌细胞内高表达)来实现对运动/肌肉收缩引起的GLUT4基因表达控制.细胞分为正常浓度葡萄糖对照组(NGLU组)、正常浓度AICAR组(NGLU AICAR组)、高浓度对照组(HGLU组)、高浓度AICAR组(HGLU AICAR组).用5 mmol/L和20 mmol/L葡萄糖浓度培养骨骼肌细胞后,NGLU AICAR组和HGLU AICAR组与肌肉收缩模拟信号刺激5-氨基-4-甲酰胺咪唑核糖核苷酸(AICAR)孵育.AICAR能激活NGLU组骨骼肌细胞AMPKα2、减少骨骼肌细胞核HDAC5蛋白、促使HDAC5与骨骼肌细胞加强因子(MEF2)蛋白分离和上调GLUT4基因的表达;相反,高浓度葡萄糖延迟由AICAR引起的AMPKα2磷酸化、AMPKα2向细胞核转入、HDAC5向细胞核转出和GLUT4基因的表达.实验结果说明在不同葡萄糖浓度下的骨骼肌细胞GLUT4基因表达变化都对应着上游AMPK蛋白和下游HDAC5蛋白的变化,AMPK可能是征用转录抑制子HDAC5来调节MEF2的活性而达到控制肌肉收缩所引起的GLUT4基因表达.     

Glucose and free fatty acid utilization during prolonged exercise in prepubertal boys in relation to catecholamine responses

Summary Ten prepubertal boys performed 60-min cycle exercise at about 60% of their maximal oxygen uptake as previously measured. To measure packed cell volume, plasma glucose, free fatty acids (FFA), glycerol and catecholamines, blood samples were drawn at rest using a heparinized cathether and at the 15th, 30th and 60th min of the exercise and after 30 min of recovery. At rest, the blood glucose concentrations were at the lowest values for normal. Exercise induced a small decrease of blood glucose which was combined with an abrupt increase of the noradrenaline concentration during the first 15 min. The FFA and glycerol concentrations increased throughout the exercise linearly with that of adrenaline. Compared to adults, the FFA uptake expressed per minute and per litre of oxygen uptake was greater in children. These results suggested that it is difficult for children to maintain a constant blood glucose concentration and that prolonged exercise provided a real stimulus to hypoglycaemia. An immediate and large increase in noradrenaline concentration during exercise and a greater utilization of FFA was probably used by children to prevent hypoglycaemia.     

Effect of insulin on sex differences in the intake of glucose solutions in rats

Adult male and female Wistar rats maintained on ad lib diet were given a choice between tap water and a solution of glucose in the concentration of either 5 or 12%. Both sexes exhibited a marked preference for glucose solutions. With the 5% solution the volume intake was similar in both sexes and the total calorie intake was normal. With the 12% solution the volume intake was higher in females than in males, while in both sexes the total calorie intake was increased to a similar (maximum acceptable) level. Treatment with Protamine Zinc Insulin (PZI) in a daily dose of 40 U/kg b.w. markedly increased the intake of the 5% solution in both sexes, but significantly more in females than in males, thus revealing sex differences which were not manifest in untreated rats. PZI treatment had little effect on 12% glucose solution intake, presumably because with this solution the total calorie intake was increased to a maximum already in untreated rats.     

Increase in insulin response to glucose in the rat chronically treated with clonidine

Summary Effect of chronic clonidine treatment on the response to glucose of rat pancreatic B-cells was investigated. Clonidine treatment was carried out for 10 days by dissolving the drug into drinking water at a concentration of 10 g/ml. Control rats were given drug-free tap water. Serum insulin responses to glucose (750 mg/kg, i. v.) of clonidinetreated rats were much smaller than those of control rats. However, after 1 day's withdrawal of clonidine, the rise in the serum insulin level induced by glucose was approximately 2-fold larger in clonidine-treated rats as compared to that in control rats. Since clonidine treatment decreased body weight of the rat by 10%–20% in 10 days, the same experiments were carried out with rats whose body weight loss was made comparable to that of clonidine-treated rats by restricting food for 10 days. Then, some animals of the group thus treated had food-restriction discontinued for 1 day. In both of the above two groups, no increment in glucoseinduced rise in serum insulin level was observed. Islets of Langerhans isolated from clonidine-treated rats showed pronounced insulin releasing capacity in response to glucose. Insulin content per islet of the clonidine-treated rat was slightly larger than that of control rat. These results indicate that the enhancement of serum insulin response to glucose following clonidine treatment is mainly attributable to the hyper-responsiveness developed in the pancreatic B-cells.     

Effects of glucose in protected ischemic kidneys

Summary Energy reserves (TAN) and anaerobic substrates (glucose, glycogen) are lower in renal than in myocardial tissue. Euro-Collins-solution contains nearly 200 mmol/l glucose, while the HTK-solution of Bretschneider contains none. Therefore the influence of glucose on kidney lactate production, on energy reserves (TAN), intrarenal pH and on morphology during the protection of ischemic kidneys was analysed using either Euro-Collins-solution, or modified Euro-Collins-solution, containing mannitol instead of glucose, or HTK-solution with and without the addition of 5, 10 and 20 mmol/l glucose. Glucose content changed during kidney perfusion with Euro-Collins-solution from about 60 to 800 mol/g_dw. While intrarenal pH decreased from 7.1 to 5.1 in Euro-Collins-kidneys during 420 min of ischemia at 25°C, pH decreased to 6.7 with the modified, mannitol containing Euro-Collins-solution. In HTK-protected kidneys intrarenal pH decreased with increasing glucose addition to the solution. Although Total Adenine Nucleotides are highest at the end of ischemia with Euro-Collins-solution, structural protection after the same ischemic stress was best in HTK-protected kidneys without glucose addition. We conclude that glucose stimulated lactate production, reduced interstitial pH in the kidney even in combination with a highly buffered solution and that it might cause greater membrane permeability leading to a structural detoriation. Mannitol seemed more appropriate than glucose in this respect, although other substances, which provide energy substrate and prevent structural damage, may exist.Supported by the Deutsche Forschungsgemeinschaft, SFB 89 —Kardiologie Göttingen     

A pharmacokinetic approach to the establishment of biopharmaceutic characteristics of different acetylsalicylic acid formulations in man

Eleven acetylsalicylic acid (ASA) formulations were administered to 26 healthy volunteers in a cross-over design. The properties of the preparations differed from conventional, effervescent, buffered to buccal. The objectives of this study were:

• 1 Consideration of the general aspects of a biopharmaceutical study: which parameter for which biopharmaceutic characteristic?
• 2 Measurement of the kinetic parameters of ASA: first-pass effect, mean residence time, mean appearance time, total body clearance, apparent volume of distribution, half-lives, etc.
• 3 Comparison of the formulations.

Most of the formulations yield mean residence times for ASA of 0.3–1.0h, which do not differ significantly (p > 0.05). For most of the products the first-pass effect is about 40 per cent; the average values of the apparent volume of distribution and whole body clearance, corrected for the first-pass effect, are about 201 and 650 ml min^?1, respectively. Peak levels are reached slowly for the buccal formulations, and rapidly for the buffered products. It is difficult, especially for ASA, to characterize the gastro-intestinal absorption with pharmacokinetic model parameters, because the first-pass effect is large and often elimination of ASA is faster than absorption. The model-independent approach has the special advantages of calculating reliable pharmacokinetic parameters, and creating theoretical possibilities to characterize the absorption patterns of the different formulations in a quantitative way. No significant differences in the values of the parameters are found between most of the formulations. The ASA first-pass effect is reasonably constant and buccal application has no advantage. Enteric coating of the outer layer of ASA formulations causes inconsistent absorption and may be categorized under ‘artificial mistakes’.     

脂肪因子与妊娠期糖尿病及糖脂代谢

妊娠期糖尿病（gestational diabetes mellitus,GDM）是妊娠期的主要并发症之一,随着我国二孩政策的实施、生活水平的提高、婚姻观念的转变,在肥胖、高龄和遗传易感性等高危因素的影响下,GDM的发病率呈逐年上升趋势。目前GDM的具体发病机制尚不明确,可能与胰岛素抵抗及胰岛β细胞功能缺陷有关。GDM孕妇体内糖代谢的紊乱会进一步导致脂质代谢的异常,从而引发不良妊娠结局。其中脂肪因子,如促代谢因子（betatrophin）、趋化素（chemerin）、C1q/肿瘤坏死因子相关蛋白（CTRP9）和瘦素（leptin）等在GDM的相关发病机制中可能起到一定作用。综述脂肪因子在调节糖脂代谢中的作用,及其在GDM病理发生中的作用。     

硒补充剂对妊娠期糖尿病患者血糖和血脂代谢及妊娠结局的影响分析

目的探讨硒补充剂对妊娠期糖尿病患者血糖、血脂代谢和妊娠结局的影响。方法将2018年7月-2019年6月盘锦市中心医院收治的45例妊娠期糖尿病患者随机分为对照组21例和硒酵母组24例。两组患者均给予健康教育、饮食指导、运动指导等常规治疗,硒酵母组在常规治疗基础上给予硒酵母片治疗。比较两组治疗前后空腹血糖、胰岛素抵抗指数、总胆固醇、甘油三酯、低密度脂蛋白胆固醇、高密度脂蛋白胆固醇水平及妊娠结局。结果治疗后两组空腹血糖和胰岛素抵抗指数均较治疗前下降,且硒酵母组两项指标均低于对照组,差异均具有统计学意义(P<0.05)。治疗后两组孕妇总胆固醇、甘油三酯、低密度脂蛋白胆固醇水平较治疗前下降,且硒酵母组三项指标水平低于对照组,差异均具有统计学意义(P<0.05)。治疗后硒酵母组高密度脂蛋白胆固醇水平高于治疗前,差异具有统计学意义(P<0.05)。对照组治疗前后以及两组治疗后高密度脂蛋白胆固醇水平差异均无统计学意义(P>0.05)。硒酵母组新生儿高胆红素血症发生率和新生儿住院率均低于对照组,差异均具有统计学意义(P<0.05)。两组其他不良妊娠结局差异均无统计学意义(P>0.05)。结论硒补充剂能够进一步改善妊娠期糖尿病患者的血糖和血脂代谢,同时,也能降低新生儿高胆红素血症发生率和住院率,值得推广使用。     

妊娠期糖尿病患者产后糖代谢异常转归影响因素分析

目的探讨妊娠期糖尿病(GDM)患者产后糖代谢异常(AGM)转归及其影响因素。方法选择2019年1月至12月,于四川大学华西第二医院孕期被诊断为GDM,并于产后4~12周进行75 g口服葡萄糖耐量试验(OGTT)筛查的1175例单胎妊娠产妇为研究对象。根据其产后糖代谢是否正常,将其分为研究组(n=361,产后AGM者)与对照组(n=814,产后糖代谢正常者)。采用回顾性分析方法,收集受试者一般临床资料及孕期与产后4~12周75 g OGTT结果等,并采用成组t检验或χ^(2)检验进行统计学分析。对GDM患者产后AGM转归相关影响因素进行单因素分析与多因素非条件logistic回归分析,探讨其AGM转归的独立影响因素。本研究遵循的程序符合病例收集医院伦理委员会制定的伦理学标准,得到该伦理委员会批准[审批文号:医学科研2021伦审批第(181)号]。结果①24~28孕周时,1175例GDM患者75 g OGTT结果提示,空腹血糖(FPG)及OGTT 1、2 h血糖指标中,1、2、3项升高者分别为639例(54.4%)、373例(31.7%)与163例(13.9%)。②产后4~12周时,1175例GDM患者75 g OGTT结果提示,产后糖代谢正常者为814例(69.3%),AGM为361例(30.7%),包括空腹血糖受损(IFG)为19例(1.6%),糖耐量受损(IGT)为294例(25.0%),IFG+IGT为23例(2.0%),疑似2型糖尿病(T2DM)患者为25例(2.1%)。③产后AGM转归影响因素的单因素分析结果显示,研究组GDM患者年龄、糖尿病家族史发生率,24~28孕周OGTT 1、2 h血糖值,以及2项血糖指标(OGTT 1、2 h血糖)均升高与3项血糖指标(FPG及OGTT1、2 h血糖)均升高者所占比例,均显著高于对照组,而研究组仅1项血糖指标(FPG或OGTT 1 h血糖)升高者所占比例,则显著低于对照组,2组比较,差异均有统计学意义(P<0.05)。④多因素非条件logistic回归分析结果:模型1将受试者年龄、糖尿病家族史及24~28孕周OGTT 1、2 h血糖值进行多因素logistic回归分析结果显示,糖尿病家族史及24~28孕周OGTT 1、2 h血糖值,均为GDM患者产后AGM转归的独立危险因素(OR=1.693、1.205、1.355,95%CI:1.208~2.373、1.088~1.335、1.204~1.524,P=0.002、<0.001、<0.001)。模型2将受试者年龄、糖尿病家族史、24~28孕周OGTT血糖指标升高项目进行多因素logistic回归分析结果显示,糖尿病家族史及24~28孕周OGTT 2项血糖指标(OGTT 1、2 h血糖)升高与3项血糖指标均升高,均为GDM患者产后AGM转归独立危险因素(OR=1.668、1.421、1.747,95%CI:1.192~2.333、1.035~1.952、1.195~2.553,P=0.003、0.030、0.004);24~28孕周仅FPG或OGTT 1 h血糖升高为其独立保护因素(OR=0.401、0.646,95%CI:0.240~0.670、0.418~0.997,P<0.001、=0.048)。结论对于GDM患者产后AGM转归,临床应关注其年龄、糖尿病家族史、孕期OGTT结果等指标。对GDM高危人群进行上述指标持续监测与规范干预,是健全GDM孕前-孕期-产后全程管理的重要环节。     

氟尿嘧啶对大鼠小肠的损伤作用研究

目的：探讨氟尿嘧啶对大鼠小肠的损伤作用,方法：通过胃管连续2d给予大鼠尿嘧啶125mk/kg.d,观察给药后第1天－第7天大鼠每日饮食量的变化,给药前和给药后的第8天分别称大鼠体重,给药后第8天测定门静脉血流量及门静脉压,分析小肠形态学变化,观察小肠对色氨酸吸收能力,测定动脉血谷氨酰胺的浓度,结果：给药后第1天－第7天大鼠饮食量均明显低于正常,尤其是第2天－第4天最为严重,第8天大鼠体重下降,小肠结构有明显损伤,门静脉血流量减少,门静脉压下降,小肠对色氨酸的吸收及动脉血谷氨酸胺浓度明显下降,与对照组相比的均有显著差异,结论：氟尿嘧啶可导致大鼠小肠结构和功能的明显障碍。