低剂量三氯化钐对实验性非胰岛素依赖型糖尿病大鼠胰岛的影响

观察了经腹腔注射低剂量三氯化钐（０．０５ｍｇ／ｋｇ）对链脲佐菌素引志的非胰岛素依赖型糖尿病（ＮＤＩＤＤＭ）大鼠胰岛形态和功能的影响。结果表明，三氯化钐治疗组糖耐量改善、血清胰岛水平长高、血清胰镐血糖素水平降低、平均单个胰岛面积和胰岛β细胞数量明显增多，肝细胞内糖原含量增多。提示低剂量三氯化钐对实验性ＮＩＤＤＭ大鼠有一定的治疗作用。     

Synthesis of （S）-2-Ethoxy-3-Phenylpropanoic Acid Derivatives and Their Insulin-Sensitizing Activity

A series of （S）-2-ethoxy-3-phenylpropanoic acid derivatives were synthesized and their insulin-sensitizing activities were evaluated in 3T3-L1 cells. Compounds 1b, 1d, 1e and 1f exhibited more potent insulin-sensitizing activity than rosiglitazone.     

二价铜离子对人胰岛淀粉样蛋白(hIAPP)(11～28)多肽聚集的影响

人胰岛淀粉样蛋白(hIAPP)与Ⅱ型糖尿病(T2DM)密切相关,被认为是导致胰岛β细胞凋亡的致病因素之一,研究发现环境因素(如金属离子、pH值和温度等)对hIAPP的聚集过程有很大影响。本文采用多种生物物理的实验方法,研究了二价铜离子对hIAPP及其片段聚集的影响。原子力显微镜(AFM)和硫代黄素T(ThT)荧光的测量表明,铜离子能够明显地抑制hIAPP(11~28)聚集成纤维,其抑制程度随铜离子浓度的增加而明显加剧。显微傅里叶变换红外光谱(Micro-FTIR)的结果表明,铜离子能够抑制hIAPP多肽中α螺旋结构向β折叠的转变。另外,氨基酸定点突变实验结果表明,hIAPP(11~28)中的组氨酸(His18)可能对多肽的聚集行为和金属铜离子的相互作用起到了决定性的影响。     

Synthesis,Kinetic and Conformational Studies of 2-Substituted-5-(β-d-glucopyranosyl)-pyrimidin-4-ones as Potential Inhibitors of Glycogen Phosphorylase

Dysregulation of glycogen phosphorylase, an enzyme involved in glucose homeostasis, may lead to a number of pathological states such as type 2 diabetes and cancer, making it an important molecular target for the development of new forms of pharmaceutical intervention. Based on our previous work on the design and synthesis of 4-arylamino-1-(β-d-glucopyranosyl)pyrimidin-2-ones, which inhibit the activity of glycogen phosphorylase by binding at its catalytic site, we report herein a general synthesis of 2-substituted-5-(β-d-glucopyranosyl)pyrimidin-4-ones, a related class of metabolically stable, C-glucosyl-based, analogues. The synthetic development consists of a metallated heterocycle, produced from 5-bromo-2-methylthiouracil, in addition to protected d-gluconolactone, followed by organosilane reduction. The methylthio handle allowed derivatization through hydrolysis, ammonolysis and arylamine substitution, and the new compounds were found to be potent (μM) inhibitors of rabbit muscle glycogen phosphorylase. The results were interpreted with the help of density functional theory calculations and conformational analysis and were compared with previous findings.     

An HPLC method for the determination of adenosine diphosphate: An important marker of hexokinase activity in metabolic diseases

Hexokinases play a critical role in the cellular uptake and utilization of glucose. As such, they are of fundamental importance to all cells. By catalyzing glucose to produce glucose‐6‐phosphate, hexokinases control the first irreversible step of glucose metabolism and initiate all major pathways of glucose consumption. Our objective was to develop and validate highly sensitive and selective high‐performance liquid chromatography with photodiode array detector (HPLC‐PDA) assays allowing the determination of adenosine diphosphate, which was used for the determination of hexokinase activity. Samples were analyzed by HPLC‐PDA using a C₁₈ analytical column (250 × 4.6 mm) for chromatographic separation. Optimal detection was achieved based on isocratic elution with a mobile phase consisting of a mixture of sodium phosphate monobasic buffer and methanol. This method met all of the requirements of specificity, sensitivity, linearity, precision, accuracy and stability generally accepted in bioanalytical chemistry and was successfully applied to a study of hexokinase activity in an alloxan‐induced diabetic rat model. Determination of hexokinase activity will permit characterization of cellular metabolic state in many diseases, such as cancer and diabetes.     

Prediction of a glucose appearance function from foods using deconvolution

The glycaemic response of an insulin-treated diabetic patientgoes through many transitory phases, leading to a steady stateglycaemic profile following a change in either insulin regimenor diet. Most models attempting to model the glucose and insulinrelationship try to model the effect of oral or injected glucoserather than that from the digestion of food. However, it isclear that a better understanding of the glycaemic responsewould arise from consideration of intestinal absorption fromthe gut. It is assumed that this type of absorption can be modelledby a so-called glucose appearance function (systemic appearanceof glucose via glucose absorption from the gut) predicting theglucose load from the food. Much research has been carried outin the areas of hepatic balance, insulin absorption and insulinindependent/dependent utilization. However, little is knownabout intestinal absorption patterns or their correspondingglucose appearance profiles. The strategy under investigation herein is to use deconvolutionor backward engineering. By starting with specific results i.e.blood glucose and insulin therapy, it is possible to work backwardsto predict the glucose forcing functions responsible for theoutcome. Assuming compartmental consistency, this will allowa clearer insight into the true gut absorption process, If successful,the same strategy can be applied to more recent glucose andinsulin models to further our understanding of the food to bloodglucose problem. This paper investigates the Lehmann-Deutsch modified model ofglucose and insulin interaction, created from the model proposedby Berger-Rodbard. The model attempts to simulate the steadystate glycaemic and plasma insulin responses, independent ofthe initial values from which the simulation is started. Glucoseenters the model via both intestinal absorption and hepaticglucose production. We considered a 70kg male insulindependentdiabetic patient with corresponding hepatic and insulin sensitivityparameters of 0.6 and 0.3 respectively. Net hepatic glucosebalance was modelled piecewise by linear and symmetric functions.A first-order Euler method with step size of 15 minutes wasemployed. For the simulation, only Actrapid and NPH injectionswere considered. The injection of insulin and the glucose fluxto the gut were started simultaneously to avoid any delay associatedwith gastric emptying. The systemic appearance of glucose was compared from two viewpoints, not only to assess the strategic principle, but alsoto assess the suitability of the modifications made by Lehmannand Deutsch. The first is a forward prediction using the compartmentalstructure. This analysis involves the rate of gastric emptyingwithout time delay. The second is a backward prediction fromexperimentally observed blood glucose profiles. Investigationsinvolved porridge, white rice and banana containing the samecarbohydrate content (25 g). Results obtained from the firstanalysis were dependent on the rate of gastric emptying, especiallyits ascending and descending branches. Results from the secondanalysis were dependent on the dose and type of insulin administered.Both predicted profiles showed consistency with physiologicalreasoning, although it became apparent that such solutions couldbe unstable. Furthermore, both types of prediction were similarin structure and appearance, especially in simulations for porridgeand banana. This emphasized the consistency and suitabilityof both analyses when investigating the compartmental accuracyand limitations within a model. The new strategic approach was deemed a success within the model,and the modifications made by Lehmann and Deutsch appropriate.We suggest that a gastric emptying curve with a possible gastricdelay is the way forward in regulating the appearance of glucosevia gut absorption. The Lehmann-Deutsch gastric curve is describedby either a trapezoidal or triangular function dependent onthe carbohydrate content of the meal. However, it was clearfrom the results obtained that carbohydrate content is onlyone factor in carbohydrate absorption, and further progressmust inevitably involve other food characteristics and propertiesif we are to improve the glucose flux.     

Rapid biochemical profiling of endothelial dysfunction in diabetes,hypertension and cancer metastasis by hierarchical cluster analysis of Raman spectra

Raman spectroscopy is a label free, versatile, simple and fast method that is increasingly used to detect pathological changes in the cells and tissues that could be useful in medical diagnostics. In this work, we tested the hypothesis that Raman spectroscopy may serve to detect endothelial dysfunction in murine models of lifestyle diseases associated with endothelial dysfunction. For that purpose, we analysed spectra from ex vivo vessels taken from mice with diabetes, hypertension and cancer metastasis. We extracted 50–70 random, single spectra, recorded in 0.2 s, from endothelium of mice with diseases and respective control animals and subjected them to hierarchical cluster analysis. Independently on the sample preparation protocol, very good discrimination was obtained for three‐tested murine models, i.e. diabetes, hypertension and cancer metastasis. Obtained sensitivity and specificity parameters were between 93% and 96% (with the exception of sensitivity in the diabetes model equalled to 88%). Our results show that single, random spectra of endothelium, recorded in less than a second, contains enough information on biochemical content of the endothelium to detect endothelial dysfunction. Furthermore, we demonstrated that biochemical profile of the endothelial dysfunction in diabetes, hypertension or cancer metastasis differs with a very high specificity and sensitivity. This conclusion can be a good starting point for the development of in vivo fast diagnostic methodology of endothelium in the future. Copyright © 2016 John Wiley & Sons, Ltd.     

Effect of nicotinamide on early graft failure following intraportal islet transplantation

Intraportal islet transplantation (IPIT) may potentially cure Type 1 diabetes mellitus; however, graft failure in the early post-transplantation period presents a major obstacle. In this study, we tested the ability of nicotinamide to prevent early islet destruction in a syngeneic mouse model. Mice (C57BL/6) with chemically-induced diabetes received intraportal transplants of syngeneic islet tissue in various doses. Islets were cultured for 24 h in medium with or without 10 mM nicotinamide supplementation. Following IPIT, islet function was confirmed by an intraperitoneal glucose tolerance test (IPGTT) and hepatectomy. The effects of nicotinamide were evaluated by blood glucose concentration, serum monocyte chemoattractant protein-1 (MCP-1) concentration, and immunohistology at 3 h and 24 h after IPIT. Among the various islet doses, an infusion of 300 syngeneic islets treated with nicotinamide exhibited the greatest differences in glucose tolerance between recipients of treated and untreated (i.e., control) islets. One day after 300 islet equivalent (IEQ) transplantation, islets treated with nicotinamide were better granulated than the untreated islets (P = 0.01), and the recipients displayed a slight decrease in serum MCP-1 concentration, as compared to controls. After 15 days, recipients of nicotinamide-pretreated islets showed higher levels of graft function (as measured by IPGTT) than controls. The pretreatment also prolonged graft survival (> 100 days) and function; these were confirmed by partial hepatectomy, which led to the recurrence of diabetes. Pretreatment of islet grafts with nicotinamide may prevent their deterioration on the early period following IPIT in a syngeneic mouse model.     

Potential role of HMG CoA reductase inhibitor on oxidative stress induced by advanced glycation endproducts in vascular smooth muscle cells of diabetic vasculopathy

Advanced glycation endproducts (AGEs)-induced vascular smooth muscle cell (VSMCs) proliferation and formation of reactive oxygen species (ROS) are emerging as one of the important mechanisms of diabetic vasculopathy but little is known about the antioxidative action of HMG CoA reductase inhibitor (statin) on AGEs. We hypothesized that statin might reduce AGEs-induced intracellular ROS of VSMCs and analyzed the possible mechanism of action of statin in AGEs-induced cellular signaling. Aortic smooth muscle cell of Sprague-Dawley rat (RASMC) culture was done using the different levels of AGEs stimulation in the presence or absence of statin. The proliferation of RASMC, ROS formation and cellular signaling was evaluated and neointimal formation after balloon injury in diabetic rats was analyzed. Increasing concentration of AGEs stimulation was associated with increased RASMC proliferation and increased ROS formation and they were decreased with statin in a dose-dependent manner. Increased NF-κB p65, phosphorylated ERK, phosphorylated p38 MAPK, cyclooxygenase-2, and c-jun by AGEs stimulation were noted and their expression was inhibited by statin. Neointimal formation after balloon injury was much thicker in diabetic rats than the sham-treated group but less neointimal growth was observed in those treated with statin after balloon injury. Increased ROS formation, subsequent activation of MAPK system and increased VSMC proliferation may be possible mechanisms of diabetic vasculopathy induced by AGEs and statin may play a key role in the treatment of AGEs-induced diabetic atherosclerosis.