线粒体的热机效率原理及其在运动疲劳中的应用

基于呼吸链电子漏现象提出了用热机效率原理描述线粒体合成ＡＴＰ 的工作效率, 指出呼吸链漏电不仅使线粒体合成ＡＴＰ 的效率降低, 而且导致线粒体生成有害的活性氧自由基, 引起线粒体损伤。通过检测游泳耗竭小鼠心肌线粒体生成过氧化氢速率的增高和线粒体呼吸对氰化钾敏感性的下降,证明了耗竭运动引起呼吸链电子漏水平明显增高。随电子漏增加而出现的活性氧的损伤表现在线粒体脂质过氧化程度增加,呼吸链四个酶复合物的活性均有不同程度降低,以及呼吸控制率的下降等等。文章讨论了呼吸链电子漏和电子漏引起的活性氧生成对线粒体合成ＡＴＰ 效率的影响。     

大鼠急性应激时肝线粒体质子跨膜转动活性的调控

大鼠烫伤早期（烫伤后３０ｍｉｎ）,肝线粒体质子加电子传递速度均加快,线粒体能化态跨膜电位降低（均以琥珀酸为底物）,线粒体膜脂流动性降低。皮下注射去甲肾上腺素后也有上述现象发生。推测急性应激通过儿茶酚胺类作用于肝细胞,导致线粒体内膜有序性增强所致。     

质子漏及其在基础代谢中的作用

“质子漏”是指电子传递链跨膜泵出的质子通过不涉及ATP合成的途径而跨膜扩散流回基质的过程,它的出现形成了由呼吸链驱动的质子泵出和质子回漏的无效循环通路.质子漏的耗氧在呼吸速率中占有重要的比重,对细胞呼吸有很强的控制作用,可以调节能量偶联系数,同时质子漏也是重要的产热过程,它承担了基础代谢产热的20%～30%.质子漏的生理功能有产热、增加代谢调节潜能、清除有害自由基和调节碳流等.     

线粒体电子传递链电子漏的化学发光测定

本实验用差速离心法分离正常大鼠肝脏和心肌线粒体 ,以lucigenin (探测超氧阴离子 )与luminol (探测过氧化氢 )为探剂 ,用化学发光法测定METC电子漏的生成。在反应体系中加入外源底物 ,其发光强度明显高于空白对照 (体系中无线粒体 )。在肝线粒体体系中 ,无论是lucigenin还是luminol诱发的发光 ,琥珀酸底物引起的发光强均要高于丙酮酸 /苹果酸引起的发光强度。在心肌线粒体 luminol体系中也有与肝线粒体相似的结果 ,在心肌线粒体 lucigenin体系中 ,加入外源底物丙酮酸 /苹果酸诱发的发光强度高于琥珀酸诱发的发光强度     

运动性疲劳状态下大鼠骨骼肌线粒体氧化磷酸化功能的研究

目的和方法 :以SD大鼠递增负荷力竭性跑台运动为运动性疲劳模型 ,分别测定运动后即刻骨路肌线粒体 :①呼吸链复合体Ⅱ Ⅲ电子传递与质子泵出比值 (H /2e) ;②以琥珀酸 (S)为底物的呼吸控制 :态 3呼吸速率(R3 )、态 4呼及速率 (R4 )、呼吸控制比 (RCR)和磷 /氧比 (P/O) ;②H ATPase合成活力 ,探讨疲劳性运动中线粒体氧化磷酸化功能改变的机理。结果 :力竭性运动后以S为底物的线粒体R4升高 2 1.10 % (P <0 .0 5 ) ;呼吸链复合体Ⅱ Ⅲ的总、净H 2e分别降低 8.5 3和 19.5 1% (均P <0 .0 5 )。底物的RCR和P/O呈显著降低 (均P <0 .0 5 ) ,而底物的R3则有所增加 (P >0 .0 5 ) ,H ATPase合成活力降低 16.68% (P <0 .0 5 )。结论 :线粒体质子漏增加 ,呼吸链电子传递与质子泵出偶联程度下降 ,氧化磷酸化脱偶联导致无效氧耗增多 ,可能是运动性疲劳状态下线粒体氧利用率下降的重要机制。     

大鼠急性应激时肝线粒体质子跨膜转运活性的调控

大鼠烫伤早期(烫伤后30min),肝线粒体质子和电子传递速度均加快,线粒体能化态跨膜电位降低(均以琥珀酸为底物),线粒体膜脂流动性降低。皮下注射去甲肾上腺素后也有上述现象发生。推测急性应激通过儿茶酚胺类作用于肝细胞,导致线粒体内膜有序性增强所致。     

绿豆线粒体呼吸链在不同电子传递途径中的电子漏

绿豆线粒体的呼喊链在氧化不同义莪时有不同的呼吸速率和电子漏速率,但是Ｏ２＾－／Ｏ２比值较稳定。呼吸链部位Ⅱ的抑制剂抗霉素Ａ对α－酮茂二酸、琥珀酸及苹果本工物时的电子漏速率和Ｏ２＾－／Ｏ２比值都明显的促进作用,说明电子漏发生的位点可能在抗纱Ａ的抑制点之前。呼吸链在氧化外源ＮＡＤＨ时,线料体所产生的地氰化物、鱼藤酮、抗弱Ａ及ＳＨＡＭ都不敏感,而对钙离子的螯合剂ＥＧＴＡ显著敏感。因此,依赖于钙离子的ＮＡ     

棕榈酸对模拟高原低氧大鼠离体脑线粒体解耦联蛋白活性及质子漏的影响

本文旨在通过观察棕榈酸对模拟高原低氧大鼠离体脑线粒体解耦联蛋白(uncoupling proteins,UCPs)活性的影响及脑线粒体质子漏与膜电位的改变,探讨UCPs在介导游离脂肪酸对低氧时线粒体氧化磷酸化功能改变中的作用.将SpragueDawley大鼠随机分为对照组、急性低氧组和慢性低氧组.低氧大鼠于低压舱内模拟海拔5 000 m高原23 h/d作低氧暴露,分别连续低氧3 d和30 d.用差速密度梯度离心法提取脑线粒体,[3H-GTP法测定UCPs含量与活性,TPMP 电极与Clark氧电极结合法测量线粒体质子漏,罗丹明123荧光法测定线粒体膜电位.结果显示,低氧使脑线粒体内UCPs含量与活性升高、质子漏增加、线粒体膜电位降低;同时,低氧暴露降低脑线粒体对棕榈酸的反应性,UCPs活性的改变率低于对照组,且线粒体UCPs含量、质子漏、膜电位变化率亦出现相同趋势.线粒体质子漏与反映UCPs活性的Kd值呈线性负相关(P<0.01 r=-0.906),与反映UCPs含量的Bmax呈线性正相关(P<0.01,r=0.856),与膜电位呈线性负相关(P<0.01,r=-0.880).以上结果提示,低氧导致的脑线粒体质子漏增加及膜电位降低与线粒体内UCPs活性升高有关,同时低氧暴露能降低脑线粒体对棕榈酸的反应性,提示在高原低氧环境下,游离脂肪酸升高在维持线粒体能量代谢中起着自身保护和调节机制.     

The Physiological Significance of Mitochondrial Proton Leak in Animal Cells and Tissues

Mitochondrial proton leak is an important component of cellular metabolism in animals and may account for as much as one quarter to one third of the Standard Metabolic Rate of the rat. The activity of the proton leak pathway is different in a wide range of animal species and in different thyroid states. Such differences imply some function for proton leak and candidates for this function include thermogenesis, protection against reactive oxygen species, endowment of metabolic sensitivity and maintenance of carbon fluxes.     

Daytime Functioning in Obstructive Sleep Apnea Patients: Exercise Tolerance, Subjective Fatigue, and Sleepiness

A sample of 32 obstructive sleep apnea patients (27 males, 5 females) was assessed with overnight polysomnography and the Multiple Sleep Latency Test (MSLT), an objective measure of daytime sleepiness. Patients also participated in a maximal exercise test, which served as an objective indicator of physical fatigue. The Fatigue Severity Scale (FSS) was used as a subjective measure of fatigue. Subjective fatigue ratings were significantly correlated with percent of predicted maximum heart rate achieved during exercise testing, suggesting that self-reported fatigue in apnea patients may refer to reduced physical fitness. FSS scores and exercise testing results were not significantly correlated with the MSLT, indicating that daytime fatigue and daytime sleepiness are independent problems in apnea patients. Participants self-reported a high level of fatigue, and exercise testing revealed decreased physical work capacity among apnea patients, but objective and subjective indicators of fatigue were not significantly correlated with apnea severity. A higher percentage of REM sleep predicted greater work capacity.     

Pluripotent Stem Cells for Uncovering the Role of Mitochondria in Human Brain Function and Dysfunction

Mitochondrial dysfunctions are a known pathogenetic mechanism of a number of neurological and psychiatric disorders. At the same time, mutations in genes encoding for components of the mitochondrial respiratory chain cause mitochondrial diseases, which commonly exhibit neurological symptoms. Mitochondria are therefore critical for the functionality of the human nervous system. The importance of mitochondria stems from their key roles in cellular metabolism, calcium handling, redox and protein homeostasis, and overall cellular homeostasis through their dynamic network. Here, we describe how the use of pluripotent stem cells (PSCs) may help in addressing the physiological and pathological relevance of mitochondria for the human nervous system. PSCs allow the generation of patient-derived neurons and glia and the identification of gene-specific and mutation-specific cellular phenotypes via genome engineering approaches. We discuss the recent advances in PSC-based modeling of brain diseases and the current challenges of the field. We anticipate that the careful use of PSCs will improve our understanding of the impact of mitochondria in neurological and psychiatric disorders and the search for effective therapeutic avenues.     

An in situ study of bioenergetic properties of human colorectal cancer: The regulation of mitochondrial respiration and distribution of flux control among the components of ATP synthasome

The aim of this study is to characterize the function of mitochondria and main energy fluxes in human colorectal cancer (HCC) cells. We have performed quantitative analysis of cellular respiration in post-operative tissue samples collected from 42 cancer patients. Permeabilized tumor tissue in combination with high resolution respirometry was used.Our results indicate that HCC is not a pure glycolytic tumor and the oxidative phosphorylation (OXPHOS) system may be the main provider of ATP in these tumor cells. The apparent Michaelis–Menten constant (K_m) for ADP and maximal respiratory rate (V_m) values were calculated for the characterization of the affinity of mitochondria for exogenous ADP: normal colon tissue displayed low affinity (K_m = 260 ± 55 μM) whereas the affinity of tumor mitochondria was significantly higher (K_m = 126 ± 17 μM). But concurrently the V_m value of the tumor samples was 60–80% higher than that in control tissue. The reason for this change is related to the increased number of mitochondria. Our data suggest that in both HCC and normal intestinal cells tubulin β-II isoform probably does not play a role in the regulation of permeability of the MOM for adenine nucleotides.The mitochondrial creatine kinase energy transfer system is not functional in HCC and our experiments showed that adenylate kinase reactions could play an important role in the maintenance of energy homeostasis in colorectal carcinomas instead of creatine kinase.Immunofluorescent studies showed that hexokinase 2 (HK-2) was associated with mitochondria in HCC cells, but during carcinogenesis the total activity of HK did not change. Furthermore, only minor alterations in the expression of HK-1 and HK-2 isoforms have been observed.Metabolic Control analysis showed that the distribution of the control over electron transport chain and ATP synthasome complexes seemed to be similar in both tumor and control tissues. High flux control coefficients point to the possibility that the mitochondrial respiratory chain is reorganized in some way or assembled into large supercomplexes in both tissues.