预耐力训练预防急性酒精性肝损伤——肝脏抗氧化能力与线粒体解偶联关系的研究

为了研究预耐力训练对急性酒精性肝损伤肝脏抗氧化能力与解偶联关系的影响,以SD大鼠建立急性酒精性肝损伤模型,以12周无负重游泳为运动手段,测定肝脏线粒体锰过氧化物歧化酶(Manganese superoxide dismutase,MnSOD)活性及线粒体三磷酸腺苷(Adenosine Triphosphate,ATP)合成活力,肝脏线粒体磷氧比(Phosphorus oxygen ratio,P/O)的变化和肝脏组织MnSOD、解偶联蛋白2(Uncoupling protein 2,UCP2)和肝细胞癌下调的线粒体转运蛋白(HCCdown-regulated mitochondrial carrier protein,HDMCP)mRNA表达.结果发现:未训练大鼠急性酒精摄入导致肝脏MnSOD酶活性、MnSOD、UCP2和HDMCP mRNA表达著升高,而ATP合成活力和P/O显著降低;预耐力训练后再给予急性酒精摄入使肝脏UCP2和HDMCPmRNA表达均显著低于直接急性酒精摄入大鼠,而肝脏线粒体P/O和MnSODmRNA表达均显著高于直接酒精摄入大鼠.结论:耐力训练可以有效提高肝脏抗氧化能力,降低解偶联蛋白表达,从而提高肝脏线粒体能量合成能力,达到预防急性酒精性肝损伤的目的.     

三种蝙蝠线粒体解偶联蛋白2（UCP2）基因的克隆和进化分析

根据几种哺乳动物UCP2基因的保守区设计一对简并引物,扩增马铁菊头蝠（Rhinolophus ferrumequinum）、长翼蝠（Miniopterus fuliginosus）和犬蝠（Cynopterus sphinx）的UCP2基因的全部编码区序列.测序结果表明,三种蝙蝠UCP2编码区全长930 bp,编码309个氨基酸,推测的氨基酸序列包含线粒体内膜载体蛋白的3个特征结构及解偶联蛋白（UCPs）的特征序列.序列分析表明,蝙蝠与其它哺乳动物UCP2的氨基酸推导序列有很高的同源性,为90.6%~97.0%.进化分析表明,UCP2基因在哺乳动物中进化过程中非常保守,受到强烈的纯化选择压力作用（ω=0.063）.Branch-specific 模型分析表明,UCP2基因在蝙蝠支系与其它不能飞行的哺乳动物、冬眠蝙蝠与非冬眠蝙蝠的进化过程所受到的选择压力无明显差异（P>0.05）.这说明在整个哺乳动物进化过程中UCP2对其能量代谢的调控均起到了重要作用.然而,UCP2如何参与哺乳动物能量调控仍有待于进一步研究.     

黄连生物碱治疗糖尿病——解偶联蛋白2

用不同浓度(4.00,2.00,1.00,0.50和0.25 mg/L)的黄连生物碱(黄连碱、小檗碱、表小檗碱、巴马汀)处理肝癌细胞(HepG2),应用HPLC和蛋白质印迹技术检测黄连生物碱在HepG2细胞中的吸收及黄连生物碱对解偶联蛋白2(UCP2)表达的影响.同一浓度下,HepG2细胞内黄连碱的浓度最高,小檗碱次之,其余生物碱含量较低;UCP2的表达量,黄连碱组最高、小檗碱次之,且随着黄连碱浓度和表小檗碱浓度的不断增加,细胞内UCP2相对表达量也增加,其它两组对UCP2表达影响不明显.黄连碱与小檗碱可以促进线粒体UCP2的表达且与治疗糖尿病并发症相关联.     

岩藻黄素降脂活性及其作用机制研究进展

岩藻黄素(Fucoxanthin)是海洋褐藻中主要的叶黄素类类胡萝卜素,也是褐藻中主要的降脂活性成分之一.岩藻黄素具有抗肿瘤、抗炎症、抗肥胖等多种生物活性,在代谢调节方面,岩藻黄素能促进脂肪分解和脂肪酸氧化,上调白色脂肪组织中线粒体解偶联蛋白1(UCP1)的表达,具有显著的抗肥胖功效,但其具体作用机制还需要进一步研究....     

运动中线粒体活性氧产生机制的研究进展

线粒体呼吸链是运动内源性活性氧(ROS)产生的主要来源,内源性ROS产生增多,可导致脂质、蛋白质及核酸等物质的氧化损伤.研究表明,ROS产生可引起质子漏,质子漏增加可降低ROS生成,提示ROS与质子漏之间存在一种反馈回路;解偶联蛋白(UCPs)参与了这种反馈调节,但其确切分子机制尚不清楚.从运动中线粒体ROS产生及其机制等方面进行了综述.     

解偶联蛋白预防肥胖、糖尿病的作用与机制

解偶联蛋白（UCPs,uncouplingproteins）是一类线粒体内膜上的载体,属于线粒体载体超家族,可以将H＋从线粒体内膜渗漏到线粒体基质中,减少ATP的合成并产生热能．了解UCPs的生理作用及调控机制可为肥胖和糖尿病的治疗提供一种新途径．     

牦牛UCP1基因生物学特性及组织表达分析

旨在克隆牦牛解偶联蛋白1(UCP1)基因序列,并对其进行生物信息学分析,进一步研究该基因在牦牛各组织的表达规律.以牦牛为实验对象,利用RT-PCR克隆得到牦牛UCP1基因全长序列,生物信息学分析牦牛CDS区,qPCR检测UCP1基因在牦牛不同组织中的表达模式.结果表明,UCP1基因全长为954 bp(GenBank N...     

褐色脂肪组织中的解偶联蛋白

褐色脂肪组织的调节产热等方面的功能近年来一直倍受关注,尤其是存在其中的解偶联蛋白更是近年来研究的热点,本文概述了解偶联蛋白及其基因的结构、解偶联蛋白的生理作用。     

线粒体外膜通道蛋白VDAC与靶向肿瘤细胞凋亡

VDAC(Voltage-dependent anion channel)是位于线粒体外膜上的一种主要通道蛋白,参与线粒体内外物质和能量的运输,在线粒体与细胞其它部位的通讯中起重要调节作用.近年来研究发现,VDAC也是线粒体与其它蛋白质相互作用的功能结合位点,可与多种凋亡调节蛋白(如HK-Ⅰ/Ⅱ、Bcl-2家族蛋白、tubulin、MAP2/4等)以及非蛋白调节因子相互作用,参与调控细胞凋亡.因此,VDAC成为线粒体凋亡通路中一种关键的靶蛋白.本文对近年来VDAC在肿瘤细胞凋亡中的作用机制进行简要综述.     

运动中的线粒体功能

综述了线粒体的生物功能.线粒体合成ATP是人体运动的主要能量来源,线粒体的功能历来受到运动科学界的重视.研究发现,运动科学中许多重要问题的研究均涉及线粒体的生物功能,包括运动中机体的疲劳发生、能量供应及其调控、运动适应以及运动中的细胞凋亡等.     

Superoxide activates mitochondrial uncoupling proteins.

Uncoupling protein 1 (UCP1) diverts energy from ATP synthesis to thermogenesis in the mitochondria of brown adipose tissue by catalysing a regulated leak of protons across the inner membrane. The functions of its homologues, UCP2 and UCP3, in other tissues are debated. UCP2 and UCP3 are present at much lower abundance than UCP1, and the uncoupling with which they are associated is not significantly thermogenic. Mild uncoupling would, however, decrease the mitochondrial production of reactive oxygen species, which are important mediators of oxidative damage. Here we show that superoxide increases mitochondrial proton conductance through effects on UCP1, UCP2 and UCP3. Superoxide-induced uncoupling requires fatty acids and is inhibited by purine nucleotides. It correlates with the tissue expression of UCPs, appears in mitochondria from yeast expressing UCP1, and is absent in skeletal muscle mitochondria from UCP3 knockout mice. Our findings indicate that the interaction of superoxide with UCPs may be a mechanism for decreasing the concentrations of reactive oxygen species inside mitochondria.     

UCP2 mediates ghrelin's action on NPY/AgRP neurons by lowering free radicals

The gut-derived hormone ghrelin exerts its effect on the brain by regulating neuronal activity. Ghrelin-induced feeding behaviour is controlled by arcuate nucleus neurons that co-express neuropeptide Y and agouti-related protein (NPY/AgRP neurons). However, the intracellular mechanisms triggered by ghrelin to alter NPY/AgRP neuronal activity are poorly understood. Here we show that ghrelin initiates robust changes in hypothalamic mitochondrial respiration in mice that are dependent on uncoupling protein 2 (UCP2). Activation of this mitochondrial mechanism is critical for ghrelin-induced mitochondrial proliferation and electric activation of NPY/AgRP neurons, for ghrelin-triggered synaptic plasticity of pro-opiomelanocortin-expressing neurons, and for ghrelin-induced food intake. The UCP2-dependent action of ghrelin on NPY/AgRP neurons is driven by a hypothalamic fatty acid oxidation pathway involving AMPK, CPT1 and free radicals that are scavenged by UCP2. These results reveal a signalling modality connecting mitochondria-mediated effects of G-protein-coupled receptors on neuronal function and associated behaviour.     

Oxidant stress evoked by pacemaking in dopaminergic neurons is attenuated by DJ-1

Parkinson's disease is a pervasive, ageing-related neurodegenerative disease the cardinal motor symptoms of which reflect the loss of a small group of neurons, the dopaminergic neurons in the substantia nigra pars compacta (SNc). Mitochondrial oxidant stress is widely viewed as being responsible for this loss, but why these particular neurons should be stressed is a mystery. Here we show, using transgenic mice that expressed a redox-sensitive variant of green fluorescent protein targeted to the mitochondrial matrix, that the engagement of plasma membrane L-type calcium channels during normal autonomous pacemaking created an oxidant stress that was specific to vulnerable SNc dopaminergic neurons. The oxidant stress engaged defences that induced transient, mild mitochondrial depolarization or uncoupling. The mild uncoupling was not affected by deletion of cyclophilin D, which is a component of the permeability transition pore, but was attenuated by genipin and purine nucleotides, which are antagonists of cloned uncoupling proteins. Knocking out DJ-1 (also known as PARK7 in humans and Park7 in mice), which is a gene associated with an early-onset form of Parkinson's disease, downregulated the expression of two uncoupling proteins (UCP4 (SLC25A27) and UCP5 (SLC25A14)), compromised calcium-induced uncoupling and increased oxidation of matrix proteins specifically in SNc dopaminergic neurons. Because drugs approved for human use can antagonize calcium entry through L-type channels, these results point to a novel neuroprotective strategy for both idiopathic and familial forms of Parkinson's disease.     

鲨鱼肝蛋白粗提物对大鼠肝线粒体抗氧化功能的影响

摘以400mg／kg2次腹腔注射硫代乙酰胺（TAA）建立大鼠急性肝损伤模型,并在注射TAA前1h以80mg／kg2次腹腔注射鲨鱼肝蛋白粗提物进行预防,研究了鲨鱼肝蛋白粗提物对大鼠肝线粒体抗氧化功能的影响。注射TAA后,大鼠肝脏线粒体腺苷二磷酸（ADP）诱导的氧消耗、呼吸控制率（RCR）、磷／氧比（P／O）、氧化磷酸化率均低于对照组,而预防组线粒体ADP诱导的氧消耗、呼吸控制率、P／O和氧化磷酸化效率均高于模型组;TAA降低了线粒体中谷胱甘肽、谷胱甘肽还原酶、谷胱甘肽过氧化物酶的水平,而鲨鱼肝蛋白粗提物则使线粒体中谷胱甘肽过氧化物酶和超氧化物歧化酶的水平明显升高,说明鲨鱼肝蛋白粗提物能部分修复线粒体受损的呼吸功能,增强线粒体抗氧化能力。     

糯谷猪解耦联蛋白3和心脏型脂肪酸结合蛋白在骨骼肌中的表达和定位

 为了探索解耦联蛋白3（uncoupling protein 3,UCP3）和心脏型脂肪酸结合蛋白（heart fatty acid binding protein,H FABP）对骨骼肌细胞中脂肪代谢的调节作用,采用RT PCR和免疫组织化学方法,研究了地方猪品种糯谷猪骨骼肌细胞中UCP3和H FABP基因的表达和细胞内定位,并与外二元杂交猪相比较,结果表明,糯骨猪的肌细胞及其细胞间隙较大,血清游离脂肪酸和肌内脂肪含量较高; 但UCP3和H FABP基因表达量和蛋白分布密度均较杂交猪低。说明糯谷猪肌细胞中UCP3和H FABP的表达量较少,肌细胞中ATP的生成效率较高,对脂肪酸供能的依赖性较低,脂肪得以贮存。提示UCP3和H FABP主要以蛋白量的变化参与猪骨骼肌细胞的脂肪代谢调节,从而影响地方猪种的肉质等性状。     

Glucose sensing by POMC neurons regulates glucose homeostasis and is impaired in obesity

A subset of neurons in the brain, known as 'glucose-excited' neurons, depolarize and increase their firing rate in response to increases in extracellular glucose. Similar to insulin secretion by pancreatic beta-cells, glucose excitation of neurons is driven by ATP-mediated closure of ATP-sensitive potassium (K(ATP)) channels. Although beta-cell-like glucose sensing in neurons is well established, its physiological relevance and contribution to disease states such as type 2 diabetes remain unknown. To address these issues, we disrupted glucose sensing in glucose-excited pro-opiomelanocortin (POMC) neurons via transgenic expression of a mutant Kir6.2 subunit (encoded by the Kcnj11 gene) that prevents ATP-mediated closure of K(ATP) channels. Here we show that this genetic manipulation impaired the whole-body response to a systemic glucose load, demonstrating a role for glucose sensing by POMC neurons in the overall physiological control of blood glucose. We also found that glucose sensing by POMC neurons became defective in obese mice on a high-fat diet, suggesting that loss of glucose sensing by neurons has a role in the development of type 2 diabetes. The mechanism for obesity-induced loss of glucose sensing in POMC neurons involves uncoupling protein 2 (UCP2), a mitochondrial protein that impairs glucose-stimulated ATP production. UCP2 negatively regulates glucose sensing in POMC neurons. We found that genetic deletion of Ucp2 prevents obesity-induced loss of glucose sensing, and that acute pharmacological inhibition of UCP2 reverses loss of glucose sensing. We conclude that obesity-induced, UCP2-mediated loss of glucose sensing in glucose-excited neurons might have a pathogenic role in the development of type 2 diabetes.     

Coenzyme Q is an obligatory cofactor for uncoupling protein function

Uncoupling proteins (UCPs) are thought to be intricately controlled uncouplers that are responsible for the futile dissipation of mitochondrial chemiosmotic gradients, producing heat rather than ATP. They occur in many animal and plant cells and form a subfamily of the mitochondrial carrier family. Physiological uncoupling of oxidative phosphorylation must be strongly regulated to avoid deterioration of the energy supply and cell death, which is caused by toxic uncouplers. However, an H+ transporting uncoupling function is well established only for UCP1 from brown adipose tissue, and the regulation of UCP1 by fatty acids, nucleotides and pH remains controversial. The failure of UCP1 expressed in Escherichia coli inclusion bodies to carry out fatty-acid-dependent H+ transport activity inclusion bodies made us seek a native UCP cofactor. Here we report the identification of coenzyme Q (ubiquinone) as such a cofactor. On addition of CoQ10 to reconstituted UCP1 from inclusion bodies, fatty-acid-dependent H+ transport reached the same rate as with native UCP1. The H+ transport was highly sensitive to purine nucleotides, and activated only by oxidized but not reduced CoQ. H+ transport of native UCP1 correlated with the endogenous CoQ content.     

Mitochondrial uncoupling protein 2 structure determined by NMR molecular fragment searching

Mitochondrial uncoupling protein 2 (UCP2) is an integral membrane protein in the mitochondrial anion carrier protein family, the members of which facilitate the transport of small molecules across the mitochondrial inner membrane. When the mitochondrial respiratory complex pumps protons from the mitochondrial matrix to the intermembrane space, it builds up an electrochemical potential. A fraction of this electrochemical potential is dissipated as heat, in a process involving leakage of protons back to the matrix. This leakage, or 'uncoupling' of the proton electrochemical potential, is mediated primarily by uncoupling proteins. However, the mechanism of UCP-mediated proton translocation across the lipid bilayer is unknown. Here we describe a solution-NMR method for structural characterization of UCP2. The method, which overcomes some of the challenges associated with membrane-protein structure determination, combines orientation restraints derived from NMR residual dipolar couplings (RDCs) and semiquantitative distance restraints from paramagnetic relaxation enhancement (PRE) measurements. The local and secondary structures of the protein were determined by piecing together molecular fragments from the Protein Data Bank that best fit experimental RDCs from samples weakly aligned in a DNA nanotube liquid crystal. The RDCs also determine the relative orientation of the secondary structural segments, and the PRE restraints provide their spatial arrangement in the tertiary fold. UCP2 closely resembles the bovine ADP/ATP carrier (the only carrier protein of known structure), but the relative orientations of the helical segments are different, resulting in a wider opening on the matrix side of the inner membrane. Moreover, the nitroxide-labelled GDP binds inside the channel and seems to be closer to transmembrane helices 1-4. We believe that this biophysical approach can be applied to other membrane proteins and, in particular, to other mitochondrial carriers, not only for structure determination but also to characterize various conformational states of these proteins linked to substrate transport.     

灯盏乙素对大鼠肝线粒体氧化损伤的抑制作用

运用Fe^2 -Vc损伤体系诱导大鼠肝线粒体氧化损伤，并研究了灯盏乙素的抑制作用，采用分光光度法测定脂质过氧化物丙二醛（MDA）含量，膜蛋白巯基含量及线粒体肿胀度，利用荧光染料Rhodamine123对线粒体进行标记，跟踪线粒体膜电位的变化，研究发现20－50μmol/L的灯盏乙素能抑制MDA的生成并促使膜巯基含量下降，对线粒体肿胀和膜电位的降低都有一定的抑制作用，并呈剂量依赖关系，结果表明灯盏乙素是一种有效的抗氧化剂，能抑制活性氧诱导的线粒体氧化损伤。μ