cDNA芯片研发及其在肥胖患者脂肪基因差异表达研究中的应用

为了加快基因功能的研究,利用已有的来源于不同组织的cDNA克隆,并通过交换和购买补充了低丰度和染色体覆盖不完全的部分cDNA,研制开发出具有相当代表性、覆盖较完全的高密度cDNA表达型基因芯片,每张芯片上含有384个质控DNA和12 630个cDNA探针,其中包括12 508个Unigene和122个表达序列标签(EST).利用这些芯片,对肥胖患者及正常人内脏脂肪组织基因表达谱进行了初步研究,并发现在肥胖患者内脏脂肪组织差异表达的基因,其中上调的有与凋亡相关的基因、与免疫有关的基因以及与能量代谢有关的基因等,而下调的主要是与脂肪酸及胆固醇合成有关的基因,对这些基因进一步的功能研究将为阐明肥胖发生机制奠定基础.     

利用生物信息学研究肥胖与2型糖尿病患者肝组织基因表达变化

目的:利用芯片数据分析工具对GEO基因芯片数据进行数据挖掘,系统分析肥胖与2型糖尿病患者肝组织相关基因表达的变化,探讨肥胖与2型糖尿病的联系及糖尿病早期预防和诊断的新靶点。方法:首先在公共芯片数据库中选择肥胖与2型糖尿病相关芯片数据(GSE15653),利用R等芯片数据分析工具分析肥胖与2型糖尿病患者肝组织基因的表达变化,并预测相关差异表达基因在血中蛋白表达。结果:肥胖患者与正常人肝组织比较发现412个差异表达基因,其中上调表达基因212个,下调表达基因200个,2型糖尿病患者中控制良好者与正常人肝组织比较发现486个差异表达基因,其中上调表达基因253个,下调表达基因233个,而2型糖尿病患者中控制不良者与正常人肝组织比较发现1051个差异表达基因,其中上调表达基因560个,下调表达基因491个;2型糖尿病控制良好者与肥胖患者肝组织有263个相同的表达变化基因,而2型糖尿病控制不良者与肥胖患者肝组织有131个相同的表达变化基因;结合蛋白质组学结果分析肥胖与2型糖尿病相关的差异表达基因中有30个蛋白表达产物是分泌型蛋白。结论:肥胖及2型糖尿病患者肝组织与正常肝组织比较基因表达均发生明显变化,其基因表达变化数目随疾病的严重性增加而增多,而且2型糖尿病的控制情况与肝组织基因表达变化有密切关系。肥胖与2型糖尿病相关的差异表达基因中表达分泌型蛋白的可进一步用于研发监测疾病发生发展的候选靶分子。     

抵抗素与肥胖和2型糖尿病

抵抗素（resistin）是近年来新发现的一个由脂肪组织特异表达分泌的细胞因子,其在前脂肪细胞分化过程中抑制脂肪生成。众多的研究显示抵抗素可诱导脂肪、肝脏及肌肉组织产生胰岛素抵抗,损伤机体的糖脂代谢功能。由于胰岛素抵抗在一些其他代谢性疾病及并发症如动脉粥样硬化及高血压发病机制中也发挥重要作用,提示抵抗素有可能介入了这些疾病的发病过程。本文简要介绍抵抗素的结构、分布及表达调控,并重点分析抵抗素在胰岛素抵抗中的作用。     

抵抗素与肥胖和2型糖尿病

抵抗素(resistin)是近年来新发现的一个由脂肪组织特异表达分泌的细胞因子,其在前脂肪细胞分化过程中抑制脂肪生成.众多的研究显示抵抗素可诱导脂肪、肝脏及肌肉组织产生胰岛素抵抗,损伤机体的糖脂代谢功能.由于胰岛素抵抗在一些其他代谢性疾病及并发症如动脉粥样硬化及高血压发病机制中也发挥重要作用,提示抵抗素有可能介入了这些疾病的发病过程.本文简要介绍抵抗素的结构、分布及表达调控,并重点分析抵抗素在胰岛素抵抗中的作用.     

利拉鲁肽联合二甲双胍对2型糖尿病合并肥胖患者胰岛β细胞功能以及内脏脂肪水平的影响

目的分析利拉鲁肽联合二甲双胍对2型糖尿病（T2DM）合并肥胖患者胰岛β细胞功能以及内脏脂肪水平的影响。 方法选取重庆两江新区第一人民医院2016年3月至2018年3月收治的194例T2DM合并肥胖患者,将其随机分为研究组、对照组,各97例,均给予为期16周的二甲双胍治疗,研究组加用利拉鲁肽皮下注射。比较两组患者治疗前后血糖、胰岛β细胞功能指数（HOMA-β）、内脏脂肪水平（VFL）等指标变化,总结利拉鲁肽在T2DM合并肥胖治疗中的临床价值。计数资料采用卡方检验,计量资料采用t检验。 结果研究组不良反应发生率为29.90﹪,对照组为23.71﹪,组间比较差异无统计学意义（χ²= 0.946,P > 0.05）。两组患者治疗后FPG、2 hPG、BMI均较治疗前下降,研究组治疗后FPG、2 hPG、BMI分别为（7.12±1.35）?mmol/?L、（9.03±2.66）?mmol/L、（26.32±1.60）kg/m²,均低于对照组的（7.83± 1.19）?mmol/L、（10.57±2.39）?mmol/?L、（27.74±1.66）kg/m²,差异有统计学意义（t = 3.886、4.241、6.066,P均?< 0.05）。两组患者治疗后HOMA-β较治疗前升高,HOMA-IR较治疗前下降,研究组治疗后HOMA-β为155.69±24.55,高于对照组的117.49±21.98,其HOMA-IR为2.30±0.71,低于后者的3.20±0.64,差异有统计学意义（t = 11.407、9.273,P均< 0.05）。两组患者治疗后VFL、脂肪率均较治疗前下降,研究组治疗后VFL、脂肪率低于对照组,且其治疗后肌肉含量较治疗前下降,差异有统计学意义（P < 0.05）。 结论在二甲双胍的基础上联合利拉鲁肽能够通过改善胰岛β细胞功能、减少内脏脂肪,达到改善T2DM合并肥胖患者的胰岛素抵抗的目的,是一种安全、有效的治疗方案。     

运动增加肥胖和2型糖尿病患者体内GLP-1水平的机制及意义

胰高血糖素样肽-1 (glucagon-like peptide-1, GLP-1)的减少在肥胖和2型糖尿病(type 2 diabetes mellitus, T2DM)中的重要作用为运动降低体重和血糖、改善胰岛素敏感性,以及防治肥胖及T2DM的机制研究提供了一个新视角。近年来的研究发现,运动可能通过谷氨酰胺(glutamine, Gln)、白介素-6 (interleukin-6,IL-6)、游离脂肪酸(free fatty acid, FFA)、交感-肾上腺髓质系统介导GLP-1的增加。增加的GLP-1可发挥改善胰岛β细胞功能,促进β细胞增殖、抑制β细胞凋亡、抑制食欲和胃排空以及降低chemerin等作用,从而提高胰岛素敏感性、减少能量摄入和改善血糖水平。这可能是运动防治肥胖、T2DM的机制之一,但仍需更多研究证实。该文就运动增加肥胖和2型糖尿病患者体内GLP-1水平的作用、机制及其生物学意义做一综述。     

腹部脂肪分布与2型糖尿病合并肥胖患者胰岛素抵抗和骨密度的关系研究

目的:探讨腹部脂肪分布与2型糖尿病合并肥胖患者胰岛素抵抗和骨密度的相关性。方法:选择2017年2月至2019年7月青岛大学附属医院内分泌与代谢病科收治的159例肥胖合并2型糖尿病患者(观察组)和同期100例单纯性肥胖且口服葡萄糖糖耐量试验(OGTT)正常者(对照组)。采用多层螺旋CT(MSCT)测量腹腔内脂肪面积(VFA)、腹内脂肪体积(IAFV)、全腹脂肪体积(TAV),并计算IAFV/TAV;采用全自动生化分析仪检测受试者血脂、空腹血糖(FPG)、空腹胰岛素(FINS)水平,并计算胰岛素抵抗指数(HOMA-IR);采用骨密度仪测量腰椎骨密度、股骨骨密度、髋骨骨密度。分析VFA、IAFV、TAV、IAFV/TAV与胰岛素抵抗和骨密度之间相关性。结果:观察组VFA、IAFV、IAFV/TAV、FPG、HOMA-IR、FINS、总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)高于对照组(P0.05),高密度脂蛋白胆固醇(HDL-C)、腰椎骨密度、股骨骨密度、髋骨骨密度低于对照组(P0.05),TAV与对照组比较差异无统计学意义(P0.05)。Pearson相关分析结果示VFA、IAFV、IAFV/TAV与FPG、HOMA-IR、FINS、TC、TG、LDL-C呈正相关(P0.05),与腰椎、股骨、髋骨骨密度、HDL-C呈负相关(P0.05),偏相关分析结果显示,VFA、IAFV、IAFV/TAV与HOMA-IR呈正相关(P0.05),与腰椎、股骨、髋骨骨密度呈负相关(P0.05)。结论:腹内脂肪异常堆积与2型糖尿病合并肥胖患者胰岛素抵抗和骨密度具有显著相关性,临床可通过检测腹内脂肪分布情况预估2型糖尿病合并肥胖患者发生胰岛素抵抗和骨质疏松的风险。     

脂肪细胞分泌蛋白质Acrp30及其生理功能

脂肪组织是人体最大的能量储存库,在需要时动用储存的脂肪,向机体供能。脂肪组织同时也是一个极其重要的分泌器官。Acrp30(adipocyte complete related protein 30 kD,)就是由脂肪组织分泌的一种活性蛋白质,参与能量代谢、炎症反应等生理过程,与肥胖、Ⅱ型糖尿病、动脉粥样硬化、心血管疾病等疾病有密切的关系。     

内脏脂肪素,一种新发现的肽类激素

内脏脂肪素（visfatin）,即：前B细胞集落增强因子（pre—B cellcolony—enhancing factor,PBEF）,是一种新发现的具有结合并激活胰岛素受体、模拟胰岛素作用的肽类激素。首先在内脏脂肪中发现,在骨髓、肝脏、肌肉等多种组织中均有表达。并具有组织特异性。内脏脂肪素具有多种生物学活性,如降血糖、模拟胰岛素样作用、增加胰岛素敏感性、参与炎症应答、延缓中性粒细胞凋亡、调节脂代谢和自分泌、旁分泌、内分泌等作用。目前内脏脂肪素在机体内的具体作用机制尚不十分清楚．但其发现可能为研究相关疾病的发病机制及治疗提供新的思路。     

苦荞主食干预对2型糖尿病伴不同程度肥胖患者体成分及血脂的影响

目的：研究苦荞作为主食对2型糖尿病伴超重或肥胖患者的体成分、血糖及血脂的影响。方法：以109名2型糖尿病伴超重或肥胖患者为研究对象,对照组54人完成4w的营养指导和健康培训,试验组55人除接受健康培训以外,采用纯苦荞食品作为主食替代部分精制米面,完成了为期4w的主食干预。干预前后进行问卷调查、体格测量并采集空腹静脉血。测定体成分、空腹血糖（FBG）、糖化血红蛋白（HbA1c）、甘油三酯（TG）、总胆固醇（TC）、高密度脂蛋白胆固醇（HDL-C）和低密度脂蛋白胆固醇（LDL-C）。结果：与仅接受营养指导和健康培训的患者相比,超重患者和肥胖患者食用苦荞后,膳食结构中蛋白质和膳食纤维的含量显著升高。同时,TC和LDL-C显著降低,且呈现显著的组间差异。此外,肥胖患者的腰围和腰臀比在干预后显著降低,且呈现显著的组间差异。结论：苦荞作为主食替代部分精制米面,每天食用量高于100g/d可以增加蛋白质及膳食纤维的摄入量,改善膳食结构,有效降低2型糖尿病伴肥胖患者的腰围,控制血液中胆固醇浓度。     

Identification of differentially expressed genes in omental adipose tissues of obese patients by suppression subtractive hybridization

To identify differentially expressed genes between obese individuals and normal control, we have undertaken suppression subtractive hybridization (SSH). Omental adipose tissues were obtained via abdominal surgery for appendicitis in both 13 obese subjects [BMI (body mass index) >30 kg/m2] and 13 normal subjects (BMI >18 and <25 kg/m2). Following SSH, about one thousand clones were sequenced and found to derive from 426 different genes. These predominately expressed genes included genes involved in lipid metabolism, cytokines, signal transduction, GLUT4 translocation, cell cycle and growth, cytoskeleton, and others. Although more detailed analyses are necessary, it is anticipated that further study of genes identified will provide insights into their specific roles in the etiology of obesity.     

Kefir prevented excess fat accumulation in diet-induced obese mice

Excessive body fat accumulation can result in obesity, which is a serious health concern. Kefir, a probiotic, has recently shown possible health benefits in fighting obesity. This study investigated the inhibitory effects of 0.1 and 0.2% kefir powder on fat accumulation in adipose and liver tissues of high-fat diet (HFD)-induced obese mice. Kefir reduced body weight and epididymal fat pad weight and decreased adipocyte diameters in HFD-induced obese mice. This was supported by decreased expression of genes related to adipogenesis and lipogenesis as well as reduced proinflammatory marker levels in epididymal fat. Along with reduced hepatic triacylglycerol concentrations and serum alanine transaminase and aspartate transaminase activities, genes related to lipogenesis and fatty acid oxidation were downregulated and upregulated, respectively, in liver tissue. Kefir also decreased serum triacylglycerol, total cholesterol, and low-density lipoprotein–cholesterol concentrations. Overall, kefir has the potential to prevent obesity.     

低氧对肥胖小鼠棕色脂肪组织相关基因表达的影响及其机制

目的：分析肥胖小鼠在低氧暴露后棕色脂肪组织的差异表达基因及通路,以探讨低氧影响棕色脂肪组织活化的机制。方法：30只雄性C57BL/6J小鼠,其中8只为普通对照组（N,n=8）;其余饲喂高脂饲料8周后,肥胖建模成功小鼠随机分为两组：肥胖对照组（OB,n=8）和肥胖低氧组（H,n=8）。H组进行11.2%氧浓度8 h/d,6天/周共4周的低氧暴露。4周后,测试血糖、血脂,取肩胛处棕色脂肪组织进行mRNA表达谱芯片扫描和生物信息学分析。利用KOBAS2.0软件对所筛选差异表达基因,并对参与关键生物过程和信号通路的差异基因进行实时荧光qPCR验证。结果：干预结束后,H组较OB组体重和血脂血糖水平显著降低;OB组较N组的上调差异基因802个,下调1 175个,差异基因的功能主要集中在糖脂合成代谢及免疫炎症反应过程;H组较OB组上调基因297个,下调228个,主要参与的生物过程有糖脂代谢、脂质转运过程、肌肉组织发育过程及脉管系统发育过程;低氧暴露调节肥胖机体棕色脂肪的通路主要集中在HIF-1、PI3K-Akt、FoxO和ErbB信号通路等过程。结论：11.2%氧气暴露可通过调节一系列棕色脂肪相关基因表达而提高棕色脂肪活性,从而下调肥胖机体体重。     

High visceral fat mass and high liver fat are associated with resistance to lifestyle intervention

Objective: High visceral adipose tissue (VAT) and high liver fat (LF) are associated with the metabolic syndrome and diabetes. We studied changes in these two fat depots during weight loss and analyzed whether VAT and LF at baseline predict the response to lifestyle intervention. Research Methods and Procedures: One hundred twelve subjects (48 men and 64 women; age, 46 ± 11 years; BMI, 29.2 ± 4.4 kg/m²) were studied after a follow up‐time of 264 ± 60 (SD) days. Insulin sensitivity was estimated from the oral glucose tolerance test. Body fat depots were quantified using magnetic resonance imaging and spectroscopy. Results: Cross‐sectionally high VAT (r = ?0.22, p = 0.02) and high LF (r = ?0.36, p < 0.0001) were independently associated with low insulin sensitivity. With intervention, BMI (?3.0%), VAT (?12.0%), and LF (?33.0%) were reduced (all p < 0.001). Insulin sensitivity was improved (+17%, p < 0.01). The changes in BMI (r = ?0.41), VAT (r = ?0.28), and LF (r = ?0.39) were associated with the increase in insulin sensitivity (all p < 0.01). High VAT (r = ?0.28, p = 0.01) and high LF (r = ?0.38, p < 0.01) at baseline were associated with a lesser increase in insulin sensitivity. Discussion: Baseline values and changes in BMI, VAT, and LF are related to changes in insulin sensitivity during lifestyle intervention. Subjects with high VAT and LF have a lower chance of profiting from lifestyle intervention and may require intensified lifestyle prevention strategies or even pharmacological approaches to improve insulin sensitivity.     

Identification of differentially expressed genes in normal and tumor human gastric tissue

The search for differentially expressed genes in gastric cancer may help define molecular alterations and molecular diagnosis of gastric cancer. Using the differential display PCR technique, we identified 18 genes that are differentially expressed between normal and tumor human gastric tissues. Their expressions were verified with reverse Northern blot analysis and Northern blot analysis. Oxidative phosphorylation-related genes, antizyme inhibitor of ornithine decarboxylase, protein phosphatase-1beta, 35-kDa peroxisomal membrane protein, and cystic fibrosis transmembrane conductance receptor were highly expressed in tumor tissue, whereas pepsinogen A, Na-K ATPase alpha subunit, nerve growth factor receptor, and alpha-tropomyosin were highly expressed in normal tissue. In addition, 3 unknown genes were found to be differentially expressed in paired gastric tissues. These differentially expressed genes may provide significant opportunities for further understanding of gastric carcinogenesis and the molecular diagnosis of gastric cancer.     

White-to-brown transdifferentiation of omental adipocytes in patients affected by pheochromocytoma

In all mammals, white adipose tissue (WAT) and brown adipose tissue (BAT) are found together in several fat depots, forming a multi-depot organ. Adrenergic stimulation induces an increase in BAT usually referred to as “browning”. This phenomenon is important because of its potential use in curbing obesity and related disorders; thus, understanding its cellular mechanisms in humans may be useful for the development of new therapeutic strategies. Data in rodents have supported the direct transformation of white into brown adipocytes. Biopsies of pure white omental fat were collected from 12 patients affected by the catecholamine-secreting tumor pheochromocytoma (pheo-patients) and compared with biopsies from controls. Half of the omental fat samples from pheo-patients contained uncoupling protein 1 (UCP1)-immunoreactive-(ir) multilocular cells that were often arranged in a BAT-like pattern endowed with noradrenergic fibers and dense capillary network. Many UCP1-ir adipocytes showed the characteristic morphology of paucilocular cells, which we have been described as cytological marker of transdifferentiation. Electron microscopy showed increased mitochondrial density in multi- and paucilocular cells and disclosed the presence of perivascular brown adipocyte precursors. Brown fat genes, such as UCP1, PR domain containing 16 (PRDM16) and β3-adrenoreceptor, were highly expressed in the omentum of pheo-patients and in those cases without visible morphologic re-arrangement. Of note, the brown determinant PRDM16 was detected by immunohistochemistry only in nuclei of multi- and paucilocular adipocytes. Quantitative electron microscopy and immunohistochemistry for Ki67 suggest an unlikely contribution of proliferative events to the phenomenon. The data support the idea that, in adult humans, white adipocytes of pure white fat that are subjected to adrenergic stimulation are able to undergo a process of direct transformation into brown adipocytes. This article is part of a Special Issue entitled Brown and White Fat: From Signaling to Disease.     

Functional classes of bronchial mucosa genes that are differentially expressed in asthma

Background  

Asthma pathogenesis and susceptibility involves a complex interplay between genetic and environmental factors. Their interaction modulates the airway inflammation and remodelling processes that are present even in mild asthma and governs the appearance and severity of symptoms of airway hyperresponsiveness. While asthma is felt to develop as the result of interaction among many different genes and signalling pathways, only a few genes have been linked to an increased risk of developing this condition.