STZ诱导大鼠1型糖尿病进行性脑萎缩的磁共振成像及组织化学研究

1型糖尿病(T1DM)是一种慢性代谢疾病,主要表现为胰岛素分泌量较正常情况下降,会对人体的多个器官和系统造成持续性的损伤．关于糖尿病的横向研究发现糖尿病患者相比于正常人存在着显著的脑萎缩,但关于糖尿病引起的脑萎缩随时间发生进行性改变的研究比较少见．实验采用腹腔注射链脲佐菌素(STZ)来诱导建立大鼠的1型糖尿病模型,运用磁共振成像(MRI)的方法对萎缩的脑区进行定位并在造模后12周和20周两个时间点对脑萎缩的程度进行对比分析,然后运用组织化学染色的方法观察在MRI上出现进行性萎缩的脑区中的神经元所发生的病理改变．MRI的结果表明：STZ诱导的T1DM大鼠相比于正常对照组大鼠出现了显著性的全脑体积、灰质体积和白质体积的萎缩,并且在多个白质脑区和灰质脑区均出现了萎缩程度随着病程的延长而逐渐加重．组织化学染色的结果发现,STZ诱导的T1DM大鼠相对于正常对照组大鼠在体感皮层、运动皮层和海马CA3区,均出现明显的神经元萎缩现象．     

基于1H NMR的代谢组学方法研究葛根芩连汤对高果糖诱导胰岛素抵抗大鼠粪便代谢组的影响

研究表明过量高果糖摄入可诱导胰岛素抵抗（IR）.葛根芩连汤（GQD）是临床常用的治疗2型糖尿病和胰岛素抵抗的中药之一,但其对果糖诱导的胰岛素抵抗的肠道菌群影响的相关研究甚少.本文采用基于核磁共振氢谱（¹H NMR）的代谢组学方法研究了GQD对高果糖诱导的胰岛素抵抗大鼠粪便代谢组的调控作用.通过连续8周给予大鼠10%果糖水喂养,成功建立胰岛素抵抗大鼠模型,GQD治疗组在果糖水喂养第5周至第8周同时给予GQD（18.2g/kg/day）灌胃.结果表明：与正常对照组相比,胰岛素抵抗模型组大鼠的饮水量、饮食量、体重和IR指数明显增加,空腹血糖值在第8周明显上升.在第8周末期,GQD治疗组的IR指数较胰岛素抵抗模型组显著降低,并且代谢紊乱得到改善：与胰岛素抵抗模型组相比,GQD治疗组胰岛素抵抗中丙酸、乙酸、琥珀酸、牛磺酸和甘油水平增加;而正丁酸、丙氨酸和谷氨酸的含量降低.这一结果表明GQD能够改善胰岛素抵抗所引起的大鼠氨基酸代谢、脂肪酸氧化和肠道微生物代谢等代谢紊乱状态.     

Ⅱ型糖尿病人与大鼠红细胞拉曼光谱学比较

使用激光共聚焦拉曼光谱仪测量正常大鼠红细胞、正常人红细胞、糖尿病STZ造模大鼠红细胞、糖尿病四氧嘧啶造模大鼠红细胞和人Ⅱ型糖尿病红细胞的拉曼光谱,应用主成分分析(principal component analysis,PCA)结合支持向量机(support vector machines,SVM)分类器对数据进行判别分析,然后采用类间距离判断两种造模方法与人Ⅱ型糖尿病的接近程度。结果发现糖尿病红细胞与正常红细胞的拉曼光谱存在明显差异,糖尿病在酰胺 ⅥCO变形振动谱带处峰高显著,并在酰胺ⅤN—H变形振动谱带处谱线出现偏移,属于磷脂的脂酰基C—C骨架1 130 cm-1谱线增强,1 088 cm-1谱线强度减弱,说明糖尿病红细胞膜的通透性增强。PCA结合SVM可以很好地区分以上5类红细胞的拉曼光谱,分类器测试结果表明分类准确度达100%。通过分别计算两种造模方法与人Ⅱ型糖尿病的类间距离,发现STZ造模法更接近人Ⅱ型糖尿病。由此得出结论：拉曼光谱法可以用于糖尿病诊断,大鼠糖尿病STZ造模法更接近人类Ⅱ型糖尿病。     

斑头雁脂肪酸的气相色谱-质谱分析

采用冷冻干燥和改进的Bligh-Dyer方法提取脂肪酸,用GC-MS测定了斑头雁腿肌、胸肌组织和卵的脂肪酸组成情况,含有脂肪酸27种,均为长链脂肪酸(C14.0以上).其中饱和脂肪酸19种,占总含量的80.577％-87.701 ％;不饱和脂肪酸8种,占脂肪酸总含量的12.298％-19.423％.在斑头雁的腿肌、胸肌和卵组织中,脂肪酸组成差异不大(P＜0.05).肌肉中含有的脂肪酸种类要多于卵组织中含有的脂肪酸种类.在斑头雁组织含有10种带有甲氧基的脂肪酸,分别占腿肌脂肪酸总量的36.175％,胸肌脂肪酸的10.401％,卵脂肪酸的45.339％.     

ICP-AES测定苦瓜籽矿质元素及GC-MS鉴定其油脂脂肪酸

苦瓜籽中活性成分的研究逐渐引起人们注意,但其矿质元素组成未见报道,其油脂脂肪酸主要成分也说法不一。文章选取海南产大苦瓜籽用电感耦合等离子体原子发射光谱仪(ICP-AES)法测定矿质元素,发现其中K,Mg和P含量最高;Cr和Zn的含量分别高达5.165和45.45μg.g-1,在植物性食品中较为罕见。以超临界CO2萃取其中油脂,得率为36.89%,碱法衍生为脂肪酸甲酯后,气相色谱-质谱联用法(GC-MS)鉴定苦瓜籽油脂中脂肪酸组成,发现其饱和脂肪酸(SFA)占36.71%,主要为硬脂酸;单不饱和脂肪酸(MUFA)仅占3.33%,主要为油酸(LA);多不饱和脂肪酸(PUFA)占59.96%,主要为α-桐酸,含量高达54.26%。明确苦瓜籽微量元素与脂肪酸组成,对进一步挖掘其药、食用价值具有重要的理论指导意义。     

糖尿病大鼠视神经病变方向性扩散加权MRI表现

利用扩散加权磁共振成像技术研究了链脲佐菌素（STZ）诱导大鼠糖尿病4周时视神经的病理改变. 测量平行和垂直于视神经方向上的水质子的表观扩散系数,分别用ADC_∥和ADC_⊥来表示. 初步结果显示模型组左右侧视神经ADC_∥没有出现一致的改变,而ADC_⊥都有下降的趋势,但没有达到统计显著性. 该结果提示STZ诱导4周后扩散成像测量参数ADC_⊥的改变可能反映了糖尿病引起的视神经轴突损伤.     

银离子络合法提取核桃油中不饱和脂肪酸

对皂化后的核桃油进行酸化获得混合脂肪酸,再对其进行甲酯化得混合脂肪酸甲酯,采用硝酸银络合萃取法提取混合脂肪酸甲酯中的不饱和脂肪酸甲酯。实验结果表明,以1mol/L氢氧化钾-无水乙醇溶液进行皂化,40%甲醇水溶液溶剂,硝酸银浓度为2mol/L,0℃的条件下络合萃取2h效果最佳,不饱和脂肪酸含量由72.3%增加到96.3%。     

微孔草新品系种子油中脂肪酸成分的比较

以气相色谱法(GC)测定了微孔草种子油中脂肪酸的含量。发现微孔草种子油中多不饱和脂肪酸含量在84.49%—85.73%之间;其中α-亚麻酸含量在13.74%—16.66%之间,微孔草新品系种子油中α亚麻酸含量均大于北微1号;γ-亚麻酸含量在6.46%—7.64%之间,微孔草新品系种子油中γ-亚麻酸含量均小于北微1号。     

新疆紫花苜蓿籽脂肪酸化学成分的分析

采用索氏提取法提取新疆紫花苜蓿籽中的脂肪酸成分,进行甲酯化处理后用气相色谱-质谱联用技术分离和鉴定脂肪酸成分的组成和含量。结果表明,从紫花苜蓿籽中共分离鉴定出11种脂肪酸成分,占总量的86.71%,主要包括9,12,15-十八碳三烯酸（37.06%）、亚油酸（25.25%）、8-十四烯酸（8.1%）、棕榈酸（10.33%）等脂肪酸,其中不饱和脂肪酸占62.31%。     

油酸和亚油酸的激光拉曼光谱

不同的不饱和脂肪酸各自具有其不同的生理功能,但常见的不饱和脂肪酸产品大部分为几种脂肪酸的混合物,故在应用前对不纯的脂肪酸产品进行组成分析是必须的。测量了不饱和脂肪酸产品组分中最常见的油酸和亚油酸的拉曼光谱,确定了各拉曼谱线的振动模归属,分析了其分子的构象特征。该结果为研究长链不饱和脂肪酸的振动能级结构及能级间跃迁等做了基础工作,丰富了有机物分子的价键数据和性质。同时详细分析比较了油酸和亚油酸拉曼光谱的差异,为定性鉴别脂肪酸产品的成分提供了一种简便有效的方法,对拉曼光谱在地沟油检测方面的应用具有重要的指导意义。     

Microstructure abnormity in the optic nerve of type 1 diabetic rats revealed by diffusion tensor imaging study

Diabetic retinopathy (DR) is one of a major complication of type 1 diabetes mellitus (T1DM) and a leading cause of blindness. Evidence of animal study has shown that it is not only a microvasucular lesion of the eye, but also a neurodegeneration disease of the visual system. However, the in vivo imaging evidence of axonal degeneration in the diabetic optic nerve is scarce. Diffusion tensor imaging (DTI) technique has been proved to be an effective tool to track the integrity of the nerve fibers in the central nervous system. In this study, type 1 diabetes was induced by intraperitoneally injecting a single dose of streptozotocin (STZ) into Sprague-Dawley rats. DTI combined with histological assessments was carried out on the optic nerve to clarify the microstructural alterations underlying DTI indices changes at 4 weeks (4 w), 8 weeks (8 w) and 12 weeks (12 w) after STZ induction. The retinal changes were analyzed by pathological evaluations at 4 weeks (4 w) and 12 weeks (12 w) after STZ induction. DTI results showed significantly decreased mean diffusivity (MD) and axial diffusivity (Da) in diabetic optic nerve compared to controls at 12 w. Atrophy in diabetic nerves was monitored by high resolution T₂-weighted images. Axonal degeneration without myelin loss of the optic nerve was confirmed by histological examination. Moreover, there are positive correlations between decreased diffusivities (MD and Da) in the optic nerve and reduced total axolemmal area. The diabetic rats showed intense glial activity since 4 w and thinning of the thickness in inner plexiform layer and nerve fiber layer at 12 w in the retina. In conclusion, DTI could in vivo monitor the progression of optic nerve degeneration in diabetes and the findings in our study would help supply axonal protection for DR in preclinical practice.     

Assessment of peripheral tissue perfusion disorder in streptozotocin-induced diabetic rats using dynamic contrast-enhanced MRI

Purpose

To assess peripheral tissue perfusion disorder in streptozotocin (STZ)-induced diabetic rats by using dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI).

Materials and Methods

A rat diabetes model was produced by intravenous injection of STZ. Diabetic rats were sustainably treated with either saline or insulin using an Alzet osmotic pump. Hind paw tissue perfusion was measured by signal intensity (SI) enhancement after gadolinium diethylenetriaminepentaacetic acid injection in DCE-MRI study and quantified using the initial area under the SI-time curve (IAUC). Peripheral tissue uptake of [¹⁴C]iodoantipyrine (IAP) was also determined as a marker of tissue blood flow for comparison with the IAUC value indicating tissue perfusion.

Results

STZ caused hyperglycemia at 1 and 2 weeks after injection. Treatment with insulin significantly alleviated hyperglycemia. At 2 weeks after STZ injection, peripheral tissue perfusion was clearly reduced in the diabetic rats and its reduction was significantly improved in the insulin-treated diabetic rats. Tissue perfusion evaluated by DCE-MRI was similar to the tissue blood flow measured by [¹⁴C]IAP uptake.

Conclusion

Our findings demonstrated that DCE-MRI can assess peripheral tissue perfusion disorder in diabetes. DCE-MRI could be suitable for noninvasive evaluation of peripheral tissue perfusion in both preclinical and clinical studies. It may also be useful for developing novel drugs to protect against diabetic vascular complications.     

A quantitative study of sodium tungstate protective effect on pancreatic beta cells in streptozotocin-induced diabetic rats

Diabetes is a major public health problem. Development of new therapies that are able to improve glycemia management, cure diabetes, and can even protect from it, are of great interest. This study investigated the protective effect of sodium tungstate against STZ-induced beta-cell damages by means of stereological methods. Sixty rats were divided into six groups: control (C), tungstate-treated control (TC), STZ-induced diabetic (D), STZ-induced diabetic rats were treated by sodium tungstate from 1 week before STZ injection (TDB), food-restricted diabetic (FRD), and diabetic rats treated with sodium tungstate 1 week after STZ administration (TDA). Stereological estimation of pancreas volume, islets volume density, volume-weighted mean islets volume and mass of beta cells, islets, and pancreas and total number of islets were done. Islets volume density, volume-weighted mean islets volume, and mass of beta cells, islets, and pancreas of TDB group was significantly higher than D, FRD and TDA groups (P < 0.001) and was comparable to controls (C and TC groups). Total number of islets, pancreas wet weight and volume did not show any significant changes between these groups (P > 0.05).Results suggested that sodium tungstate preserves pancreatic beta cells from STZ-induced damages and diabetes induction in rats.     

Antidiabetic activity of Armillaria mellea polysaccharides: Joint ultrasonic and enzyme assisted extraction

Armillaria mellea polysaccharides (AMPs) were obtained by ultrasonic assisted extraction (U), enzyme assisted extraction (E) and ultrasonic-enzyme assisted extraction (UE), respectively. The yield of UE-AMPs (6.32 ± 0.14%) was 1.64 times higher than that of U-AMPs (3.86 ± 0.11%) and 1.21 times higher than that of E-AMPs (5.21 ± 0.09%); meanwhile, the highest total sugar content and the lowest protein content were found in UE-AMPs. AMPs obtained from the three extraction methods had the same monosaccharide composition but in different proportions, allowing UE-AMPs to have the most potent antioxidant activity. The antidiabetic activity of UE-AMPs was investigated in streptozotocin (STZ)-induced diabetic mice. UE-AMPs, when given by gavage, greatly prevented weight loss, increased water intake, and considerably decreased blood glucose levels in diabetic mice, which were dose-dependent (P < 0.05). In addition, UE-AMPs also had a positive effect on the reduction of lipid levels in the blood, oxidative damage and liver function impairment. The pathological observation by hematoxylin-eosin staining (HE) revealed that UE-AMPs protected the organs of mice from diabetic complications (liver disease and nephropathy). Hence, our findings demonstrate that UE-AMPs are a suitable choice for improving diabetes and its complications and have great application prospects in the fields of natural medicine and functional food.     

Ⅱ型糖尿病肾病小鼠肾水提物代谢组学研究

应用基于核磁共振的代谢组学方法研究Ⅱ型糖尿病模型db/db小鼠已发展到糖尿病肾病时肾脏代谢表型的变化.结合偏最小二乘法-判别分析(PLS-DA),结果显示,与正常组相比,模型组肾水提物中的乳酸和糖等代谢物的含量显著升高,而谷氨酸、乙酸、胆碱和甘氨酸的含量显著降低,缬氨酸、肌酐、尼克酰胺、苯丙氨酸和酪氨酸含量略有降低.实验结果表明糖尿病肾病动物模型db/db小鼠的代谢特征不同于正常小鼠.代谢组学分析方法作为辅助手段,为糖尿病肾病的发病机制提供了良好的数据支持.     

Magnetic resonance imaging (MRI) and pathophysiology of the rat kidney in streptozotocin-induced diabetes.

Proton magnetic resonance imaging was performed on rats before induction of diabetes with streptozotocin (STZ) and at 2 and 12 days postinduction. Images revealed an increase in maximal longitudinal and axial dimensions of the kidneys at 2 days and a further increase at 12 days. Similarly, an increase in the size of the remaining kidney was seen in a rat which underwent uninephrectomy as a positive control. Two major differences were observed between the kidney undergoing compensatory hypertrophy and those developing diabetic nephropathy: (i) Expansion of the renal vasculature was seen only in images of the diabetic rat; (ii) A loss in conspicuity of the normal corticomedullary junction was seen in the T2-weighted images of the diabetic rat but not in the uninephrectomized rat. Histologic examination revealed that the medulla increased to a size greater than the cortex during diabetic nephropathy whereas the medullary volume was less than that of the cortex during compensatory hypertrophy. In vitro T1 relaxation times in cortex, outer medulla and inner medulla of kidneys from control rats were measured and compared with the same respective regions in diabetic rats. When these values were correlated with tissue water content, a linear increase in relaxation rate versus percent water content from cortex to inner medulla was found in the control kidneys, but this correlation was absent in diabetic nephropathy. These studies demonstrate that MRI is an effective noninvasive tool for studying the course of renal hypertrophy and hydration changes in the development of renal disease in STZ-induced diabetes in the rat.     

Atmospheric plasma discharge sterilization effects on whole cell fatty acid profiles of Escherichia coli and Staphylococcus aureus

As an alternative to conventional sterilization methods the plasma ions created from high voltage produce radicals which have a direct impact on microorganisms in water. Cell membrane fluidity is an important factor in survival as the fluidity allows for essential cell functions in which pH and fatty acids play a major role. This study investigated the effect of plasma application upon pH and fatty acid profiles of Escherichia coli and Staphylococcus aureus. Results showed that plasma sterilization is not predominantly a result of changes to pH levels or fatty acid contents in the bacterial cell membrane.     

Label‐free optical detection of type II diabetes based on surface‐enhanced Raman spectroscopy and multivariate analysis

Surface‐enhanced Raman scattering (SERS) spectroscopy was first employed to detect oxyhemoglobin (OxyHb, the common type of hemoglobin) variation in type II diabetic development without using exogenous reagents. Using silver nanoparticles as SERS‐active substrate, high‐quality SERS spectra are obtained from blood OxyHb samples of 49 diabetic patients and 40 healthy volunteers. Tentative assignment of the observed SERS bands indicates specific structural changes of OxyHb molecule in diabetes, including heme transformation and globin variation. Furthermore, partial least squares and principal component analysis combined with linear discriminate analysis diagnostic algorithms are employed to analyze and classify the SERS spectra acquired from diabetic and healthy OxyHb, yielding the diagnostic accuracies of 90.0% and 95.5%, respectively. This exploratory work suggests that the silver nanoparticles‐based OxyHb SERS method in combination with multivariate statistical analysis has great potential for the label‐free detection of type II diabetes. Copyright © 2014 John Wiley & Sons, Ltd.