邻羟基苯磺酰苯胺类药物的解偶联活性测定

采用氧电极技术测定药物浓度与线粒体呼吸控制率(RCR)关系,及测定线粒体呼吸过程中无机磷浓度的变化量两种方法,检测新合成的5个邻羟基苯磺酰苯胺类有机化合物的活性.结果表明,它们在较低浓度范围(10-4～10-7 mmol/L)内有较强的解偶联活性.     

低氧适应的线粒体调控机制研究进展

氧气对于生物体不可或缺,当细胞面临低氧胁迫时通过诱导低氧基因做出应答。长期以来线粒体都被认为是氧浓度感受器,其呼吸链为低氧信号产生所必需,但具体机制仍不清楚,可能通过呼吸链产生ROS调控PHD活性或ROS、NO共同作用蛋白酪氨酸硝化反应产生低氧信号;线粒体基因突变和细胞代谢通路改变、呼吸链效率调控也在机体低氧适应过程中发挥重要作用。本文对线粒体如何参与生物低氧适应的机制进行综述,以期引起更多研究者对生物低氧适应的关注和对线粒体更深入系统的研究。     

硫酸铜日粮对肉鸡肝线粒体呼吸复合物活性的影响

采用玉米-豆粕型饲粮研究铜(源于硫酸铜)对肉鸡肝细胞线粒体呼吸链复合物活性的影响。在对照组(Cu 11 mg/kg,称试验组A)基础上设高铜组3个(含Cu分别为110,220和330 mg/kg,分别称为试验组B、C、D),试验期60 d。在12,24,36,48,60日龄采集肝脏,提取线粒体,测定线粒体复合物Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ活性变化。结果表明,A组和B组线粒体复合物Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ活性呈增加趋势;C组和D组活性有减弱表现,尤其是48 d之后,比各组的试验前期下降明显(P<0.05)。说明高铜日粮在一定程度上对线粒体复合物活性是有利的,但过高的铜添加日粮(≥220 mg/kg)可降低肉鸡肝细胞线粒体呼吸链复合物功能活性,可以推测线粒体是机体铜过量状态下侵害的靶部位之一,而且能作用于线粒体各个复合物,造成线粒体呼吸功能减弱,呼吸链中电子传递发生故障,从而影响线粒体功能的正常发挥。     

谷胱甘肽对铜孵育的肉鸡肝细胞线粒体的保护作用

向即时分离的肉鸡肝细胞线粒体培养液内加入终浓度分别为10、30、50μmol/L的Cu2＋和200μmol/L GSH＋50μmol/L Cu2＋,孵育20min后,观察培养液中不同铜浓度对线粒体H2O2生成速率和呼吸链复合物活性的影响。结果表明,30μmol/L和50μmol/L的Cu2＋能引起线粒体H2O2生成速率和呼吸链复合物活性出现明显加快和降低（P〈0.05或P〈0.01）,10μmol/L Cu2＋组没有出现明显变化;谷胱甘肽可显著抑制50μmol/L Cu2＋引起的变化（P〈0.01）,对线粒体具有保护作用。     

高铜日粮对肉鸡肝细胞线粒体呼吸功能的影响

采用玉米-豆粕型日粮研究铜(源于硫酸铜)对肉鸡肝细胞线粒体呼吸功能的影响.在对照组(Cu 11 mg/kg)基础上设3个高铜组(110 mg/kg、220 mg/kg和330 mg/kg,分别对应为:试验组Ⅰ、Ⅱ、Ⅲ),试验60 d,在不同时间点(12、24、36、48、60日龄)采集肝脏提取线粒体.随着饲养时间的延长,对照组和试验I组肝脏线粒体呼吸功能有增强的趋势.在48日龄之前,试验Ⅱ和Ⅲ组肝脏线粒体呼吸功能也有增强的趋势,但幅度较小,48日龄以后,肝脏线粒体呼吸功能呈现下降趋势,特别是60日龄的试验Ⅲ组.高铜日粮能影响肉鸡肝细胞线粒体的呼吸功能,日粮铜浓度为110mg/kg时,能增强线粒体的呼吸功能,但较高的铜浓度日粮则损害线粒体的呼吸功能. 、220 mg/kg和330 mg/kg,分别对应为:试验组Ⅰ、Ⅱ、Ⅲ),试验60 d,在不同时间点(12、24、36、48、60日龄)采集肝脏提取线粒体.随着饲养时间的延长,对照组和试验I组肝脏线粒体呼吸功能有增强的趋势.在48日龄之前,试验Ⅱ和Ⅲ组肝脏线粒体呼吸功能也有增强的趋势,但幅度较小,48日龄以后,肝脏线粒体呼吸功能呈现下降趋势,特别是60日龄的试验Ⅲ .高铜日粮能影响肉鸡肝细胞线粒体的呼吸功能,日粮铜浓度为110mg/kg时,能增强线粒体的呼吸功能     

不同海拔牦牛骨骼肌线粒体呼吸链复合酶活性及氧化应激指标的差异分析

线粒体呼吸链是实现氧化磷酸化的分子机构,被广泛证实是活性氧产生的主要来源,活性氧的积累会造成细胞氧化应激,因此为了阐明不同海拔地区牦牛骨骼肌线粒体呼吸链复合酶活性及抗氧化能力的差异,试验从高(海拔4 200 m)、中(海拔3 200 m)、低(海拔1 900 m)海拔地区随机选择临床健康成年雄性牦牛各5头(分别分为高、中、低海拔组),屠宰后取骨骼肌采用比色法测定5种线粒体呼吸链复合酶和抗氧化关键酶的活性及氧化应激指标。结果表明：随着海拔的升高,牦牛骨骼肌线粒体呼吸链复合酶Ⅰ～Ⅴ活性均表现为逐渐降低的趋势,其中高海拔组牦牛骨骼肌线粒体呼吸链复合酶Ⅰ、Ⅲ、Ⅳ活性与中、低海拔组牦牛差异显著(P<0.05);低海拔组牦牛骨骼肌线粒体呼吸链复合酶Ⅱ、Ⅴ活性与中、高海拔组牦牛差异显著(P<0.05)。随着海拔的升高,牦牛骨骼肌中羟自由基、丙二醛(MDA)含量和谷胱甘肽过氧化物酶(GSH-Px)活性逐渐下降,且各组间显著差异(P<0.05);总抗氧化能力(T-AOC)先升高后降低,中海拔组牦牛骨骼肌显著高于低海拔组和高海拔组(P<0.05);超氧化物歧化酶(SOD)活性先下降...     

高铜日粮对肉鸡肝细胞线粒体呼吸链复合物活性的影响

在对照组（Cu 11mg／kg，A组）基础上设高铜组3个（110mg／kg、220mg／kg和330mg／kg，分别对应为试验组B、C、D），试验期为60d，在12、24、36、48、60日龄采集肝组织，提取线粒体，测定线粒体复合物Ⅰ、Ⅱ、Ⅲ、Ⅳ和Ⅴ的活性变化。结果表明，11mg／kg和110mg／kg铜日粮添加组线粒体复合物Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ的活性呈增加趋势。220mg／kg和330mg／kg铜日粮添加组肉鸡肝细胞线粒体复合物Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ活性减弱，但主要在试验后期，尤其是48日龄之后，比对照组和各组内的试验前期下降明显（P〈0．05）。说明高铜日粮在一定程度上对线粒体复合物活性是有利的，但过高的铜添加到日粮中（≥220mg／kg）可降低肉鸡肝细胞线粒体呼吸链复合物的功能活性。据此可以推测，线粒体是机体铜过量状态下侵害的靶部位之一，而且能作用于线粒体各个复合物，造成线粒体呼吸功能减弱，呼吸链中电子传递发生故障，从而影响线粒体功能的正常发挥。     

TRPM2离子通道在H9N2流感病毒感染小鼠肺微血管内皮细胞线粒体损伤中的作用

旨在探讨瞬时电位受体M2离子通道(TRPM2)在H9N2流感病毒感染小鼠肺微血管内皮细胞(PMVEC)导致线粒体损伤过程中的作用。在已建立TRPM2 shRNA PMVEC的基础上,采用5MOI H9N2猪流感病毒感染细胞,在病毒作用后24和48 h提取各组细胞的线粒体进行蛋白定量,检测各组细胞线粒体超氧化物歧化酶(SOD)、谷胱甘肽(GSH)、一氧化氮合成酶(NOS)、线粒体呼吸链复合物Ⅳ活性和ATP酶水平;利用激光共聚焦显微镜观察线粒体膜电位变化(JC-1染色)及细胞凋亡(Annexin V-FITC/PI双染色法)情况。结果表明:H9N2-SIV感染后TRPM2 shRNA PMVEC内SOD活性、GSH水平和Na~+-K~+-ATP、线粒体呼吸链复合物Ⅳ活性显著高于对照shRNA PMVEC(P0.05或P0.01);mtNOS活性则极显著低于shRNA PMVEC(P0.01)。JC-1染色显示TRPM2-siRNA PMVEC线粒体膜电位水平高于对照shRNA PMVEC,但是AnnexinV-FITC/PI染色显示细胞凋亡则低于H9N2感染对照shRNA PMVEC。结果提示:TRPM2基因沉默有效减缓H9N2感染导致NOS产生,显著降低SOD、GSH、线粒体呼吸链复合物Ⅳ、Na~+-K~+-ATP的消耗以及线粒体膜电位的下降程度,进而有效缓解PMVEC线粒体损伤及细胞凋亡。     

低氧环境下菊苣酸对SD大鼠心肌组织低氧适应性的影响

旨在研究低氧环境下不同浓度菊苣酸(chicoric acid, CA)对SD大鼠心肌组织低氧适应性的影响。选取60只6～8周龄、体质量为(192.00±7.35) g的健康雄性SD大鼠(P<0.05),随机分为对照组、不同浓度CA组(10,20,40 mg/kg),每组15只，连续灌胃49 d。灌胃50 d时采集SD大鼠心脏，并测定各组大鼠心肌组织活性氧(ROS)含量，抗氧化指标超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化物酶(CAT)活性，线粒体功能相关指标线粒体呼吸链复合体Ⅰ、Ⅲ活性，能量代谢相关指标Na⁺-K⁺-ATP酶、Ca²⁺-ATP酶活性及低氧适应基因缺氧诱导因子1α(HIF-1α)、促红细胞生成素(EPO)、血管内皮生长因子(VEGF)的mRNA及蛋白表达情况。结果显示：与对照组相比，不同浓度CA组显著降低ROS活性(P<0.05);显著降低MDA含量(P<0.05),显著提高SOD、GSH-Px、CAT活性(P<0.05);显著提高线粒体呼...     

铜对肉鸡肝细胞线粒体膜通透性及呼吸的影响

为进一步了解铜对生物体的生物学效应，选用硫酸铜作为铜源，向即时分离的肉鸡肝细胞线粒体培养液内加入终浓度分别为5、10、20μmol／L的铜（以铜计），孵育10min后，观察肝细胞线粒体膜通透性及呼吸的变化。结果显示，铜对线粒体膜通透性转换孔有较显著的影响，造成线粒体不同程度的肿胀，表现为线粒体悬液在540nm处的吸光值下降，且铜浓度越高，吸光值下降越明显，在孵盲6min后3种浓度的吸光值下降百分比差异显著（P〈0．05）。随着铜浓度的升高，线粒体呼吸逐渐减弱，RCR、P／O、OPR均显著降低（P〈0．05），这些变化可能是微量元素铜对生物机体产生毒害作用的本质原因之一。     

Anthelmintic action—A metabolic approach (a review)

The effects of anthelmintic drugs on intermediary metabolism of parasites can be examined using techniques developed for the investigation of metabolic regulation. Most attention has been paid to the pathways of respiratory metabolism in parasites, since maintenance of high energy levels is essential to an organism's survival. Respiratory metabolism is thus a prime target for anthelmintic attack. Parasites differ from their hosts in a number of pathways of respiratory metabolism and they produce incompletely-oxidised organic acids as end products.A variety of anthelmintic drugs have been shown to inhibit some enzymes of these pathways, including phosphoenolpyruvate carboxykinase, malate dehydrogenase, and the fumarate reductase, ‘succinoxidase’ and succinate decarboxylase systems. Energy synthesis in mitochondria from parasites is also inhibited by many anthelmintics: this target is a particularly important one since its properties differ significantly from those of the host. This process requires considerable study and it may not necessarily be valid to equate ‘uncoupling’ in mammalian tissues with that in parasite tissues. Changes in ATP levels affect flux through the respiratory pathways; a complete assessment of effects of anthelminstics on the respiratory pathways of parasites would include measurement of the uptake and utilization of carbon sources (e.g., glucose and glycogen), of metabolic intermediates, including adenine nucleotides, and of excreted end products. Sites of change can then be identified and studied in greater detail.Comparison of the effect of mebendazole in vivo and in vitro on unrelated species, Moniezia expansa and Fasciola hepatica adults, sheds some light on the mechanism of its anthelmintic effects. This drug elicits similar changes in proportions and totals of adenine nucleotides in the two species, but it differs in the rapidly of its effect, its effect on glucose uptake and glycogen depletion, and in changes in respiratory end product. Consequently mebendazole may not act in the same way against these two species. It also affects a number of other systems in other parasites: the suggestion of a single ‘mode of action’ may be inappropriate for this compound.Respiratory metabolism is not the only system affected by anthelmintics: other parasite systems are also attacked, but the final effect of lethal anthelmintic is observed as changes in the respiratory metabolic pathways. Some anthelmintics, for example, levamisole, affect the respiratory pathway directly, but also affect other systems (the nervous system in the case of levamisole) and may be effective in vitro but not in vivo. In these cases it is necessary to distinguish between effects that may be subordinate to a primary effect, and to take host and permeability factors into account.     

The relationships among mitochondrial uncoupling protein 2 and 3 expression, mitochondrial deoxyribonucleic acid single nucleotide polymorphisms, and residual feed intake in Angus steers

The objective of this study was to determine the relationships of uncoupling protein 2 and 3 expression, SNP of mitochondrial DNA, and residual feed intake (RFI) in Angus steers selected to have high or low RFI. Individual feed intake was measured via the GrowSafe feed intake system over a 3-mo period and used to calculate RFI, a measure of efficiency. Based on these calculations, 6 low- (average RFI = -1.57 kg) and 6 high- (average RFI = 1.66 kg) RFI steers were selected for further study. Blood was collected via jugular venipuncture 1 wk before slaughter for the isolation of mitochondrial DNA. The steers were then killed to collect LM for the measurement of uncoupling protein 2 and 3 mRNA and protein expression. Protein and mRNA expression of uncoupling protein 2 and 3 were determined by Western blotting and quantitative PCR, respectively. To determine SNP of mitochondrial DNA, total DNA was isolated from blood via standard phenol/chloroform extraction; fragments were amplified with PCR and sequenced with an automated nucleotide sequencer. Average daily gain and carcass composition were not different (P > 0.13) between the high- and low-RFI steers; however, ADFI by the high-RFI animals was 3.77 kg greater (P < 0.001) than the low-RFI animals. No difference (P > 0.55) was observed between the high- and low-RFI animals in their expression of uncoupling protein 2 or 3 mRNA or protein. On average 9.8 and 8.9 polymorphisms were found per mitochondrial genome for the low- and high-RFI steers, respectively. None of these polymorphisms were related to RFI. It seems that the expression of uncoupling protein 2 and 3 and mitochondrial DNA sequence are not related to RFI status.     

动物解耦联蛋白基因的同源性分析及对能量代谢的作用

文中分析比较11种物种解耦联蛋白(UCPs)基因核苷酸序列同源性,综述了UCPs产热机制和UCPs基因在脂肪和肌肉组织中的表达调控。为深入开展UCPs对能量代谢调控方面的研究奠定基础。     

Respiration and motility of epididymal sperm from slaughtered bulls

The simple method of retrograde flushing of spermatozoa from the epididymal cauda of slaughter bulls yielded an average of 2 x 10(9) spermatozoa from one cauda. The plasma membrane was intact in something between 80 and 85 percent of all epididymal spermatozoa. Respiratory rates and motility were clearly below values typical of ejaculated spermatozoa, though sizeable differences were found to exist for both parameters between individual preparations. The endogenous substrates available proved to be sufficient for such low metabolic activity, whereas higher energy turnover required the presence of exogenous substrates, such as lactate, pyruvate or fructose. Respiratory capacity, determined by uncoupling of oxidative phosphorylation in the presence of lactate, was comparable to that of ejaculated bull spermatozoa. The above findings are likely to suggest that the lower respiratory rates of epididymal spermatozoa were attributable to the fact that their consumption for motility of adenosine triphosphoric acid (ATP) was lower than that of ejaculated spermatozoa.     

抗肝片吸虫病药物化合物：N-（2＇，4＇，5＇-三氯）苯基-2-羟基-3，5，6-三氯苯磺酰胺的杀虫活性研究

本文研究了治疗牛、羊等家畜肝片吸虫病药物N-（2＇，4＇，5＇-三氯）苯基-2羟基-3，5，6-三氯-苯磺酰胺化合物的杀虫机理，通过实验及药物分子的结构分析，初步认为其杀虫机理是对线粒体氧化磷酸化反应的解偶联作用；采用氧电极法、测线粒体呼吸过程中磷的变化以及测ATP酶活性等方法研究其对线粒氧化磷酸化反应的解偶联作用，结果表明标题药物化合物是较好的解偶联剂，其解偶联活性高于典型的解偶联剂DNP。     

PRDM16——控制BAT和WAT发育的分子开关

棕色脂肪组织通过大量表达解偶联蛋白-1(UCP-1)以加强线粒体呼吸作用的解偶联,使能量以热量形式散失,从而抵御肥胖和相关疾病.研究发现,PRDM16是调节棕色脂肪细胞形成的关键转录因子,是控制棕色和白色脂肪细胞特异基因表达程序的分子开关,其机制是C末端结合蛋白和过氧化物酶体增殖剂活性受体γ辅助激活子1 (PPARγ-coactivator-1α and β,PGC-1α/β) 竞争性结合PRDM16,分别形成PRDM16/CtBPs和PRDM16/PGC-1s转录复合物,前者可激活棕色脂肪特异基因的表达,后者则抑制白色脂肪特异基因的表达.论文通过对PRDM16的结构、功能及其作用机制的介绍,有助于研究者了解棕色脂肪组织发育的调控系统,为肥胖和相关疾病的治疗提供新思路.     

Selection for high and low oxygen consumption altered hepatic mitochondrial energy efficiency in mice

Selection for high (H) and low (L) oxygen consumption (OC) as an indirect estimation of maintenance energy requirement was determined. Feed intake and body weight were measured and feed conversion ratio (FCR) of 4–8‐week‐old mice was calculated. Respiratory activity of hepatic mitochondria was measured at 12 weeks. Total feed intake (H: 103.74 g, L: 97.92 g, P < 0.01), daily feed intake (H: 3.70 g/day, L: 3.50 g/day, P < 0.01) and FCR (H: 18.79, L: 15.50, P < 0.01) were significantly different between lines. The line by sex interaction was significant for FCR. No line differences were observed in males; and the FCR of the H line was greater than in the L line in females. H line mice had the highest hepatic mitochondrial respiratory activity in state 2 (P < 0.01), the highest uncoupled respiratory rate of mitochondria in the presence of an uncoupling agent (P < 0.001), and the mitochondrial proton leak. The adenosine diphosphate/ O ratio was highest in the L line (P < 0.05). This suggests that the selection for high and low OC induced differences in basal mitochondrial respiration and basal metabolism, resulting in difference in FCR between H and L lines.