线粒体通透性转换孔、细胞色素C与细胞凋亡

线粒体通透性转换孔（permeability transition pore,PTP)开放与细胞色素C从线粒体释放是细胞凋亡中的两个关键事件;有关细胞色素C释放的机制以及PTP开放是否是细胞色素C释放的重要或唯一途径仍需深入探索。     

活性氧、线粒体通透性转换与细胞凋亡

线粒体是真核细胞中非常重要的细胞器,细胞中的活性氧等自由基主要来源于此,线粒体膜的通透性转换(mitochondrial permeability transition,MPT)及其孔道(mitochondrialpermeability transition pore,MPTP)更是在内源性细胞凋亡中发挥了关键作用。持续性的线粒体膜通透性转换在凋亡的效应阶段起决定性作用,可介导细胞色素c等促凋亡因子从线粒体释放到胞浆中,进一步激活下游的信号通路,导致细胞不可逆地走向凋亡。瞬时性的线粒体膜通透性转换及其偶联的线粒体局部的活性氧爆发同样具有促凋亡的作用。线粒体通透性孔道的开放释放出大量活性氧,这些活性氧又能够进一步激活该孔道,以正反馈的形式进一步加剧孔道的打开,放大凋亡信号。活性氧、线粒体通透性转换与细胞凋亡之间具有密不可分的联系,本文根据已知的研究结果集中讨论了这三者的关系,并着重论述了该领域中的最新发现和成果。     

线粒体通透性转换孔在缺血预处理脑保护中的作用及机制

目的：线粒体通透性转换孔通透性改变是导致缺血再灌注损伤的原因,线粒体功能的致命性改变最终引起细胞凋亡,本研究旨在观察线粒体通透性转换孔（mitochondrial permeability transition pore,MPTP）在缺血再灌注及缺血预处理脑保护中的作用;方法：将体外培养8天的海马神经元细胞分为五组,正常对照组（A组）,缺血再灌注组（B组）,缺血预处理＋缺血再灌注组（C组）,苍术苷＋缺血再灌注组（D组）,缺血预处理＋苍术苷＋缺血再灌注组（E组）。使用流式细胞术检测各组细胞凋亡率,罗丹明123染色流式细胞术检测线粒体膜电位,Western-blot检测Bcl-2,Bax的表达。结果：与A组比较,其余四组线粒体膜电位均降低,神经元凋亡率升高（P〈0．05）;与B组比较,c组线粒体膜电位升高,神经元凋亡率升高,Bcl-2表达上调,Bax表达下调（P〈0．05）;与c组比较,E组粒体膜电位降低,神经元凋亡率升高,Bcl．2表达下调,Bax表达上调（P〈0．05）。结论：我们在细胞及分子生物学水平对MPTP及缺血预处理的研究后发现,缺血预处理能有效减轻海马神经元缺血再灌注损伤,抑制缺血再灌注后神经细胞凋亡,其机制与抑制MPTP的开放有关。     

线粒体通透性转换孔在缺血后处理中的作用和机制

心肌细胞急性缺血后,及时再灌注能够挽救缺血心肌细胞的活力、减少梗死面积、促进心肌细胞功能恢复。但是再灌注是一把“双刃剑”,它产生大量活性氧类（reactive oxygen species,ROS）和Ca2＋超载,开放线粒体通透性转换孔（mitochondrial permeability transition pore,mPTP）,使线粒体肿胀,外膜破裂导致心肌细胞坏死。mPTP是线粒体非特异性的转换孔,由电压依赖性阴离子通道（voltage-dependent anion channel,VDAC）、腺苷酸转位蛋白（adeninenucleotide translocator,ANT）和亲环蛋白D（cyclophilin D,CYPD）组成。mPTP关闭维持线粒体结构完整,是缺血心肌细胞功能恢复的先决条件。缺血后处理通过减少再灌注早期ROS大量释放和拮抗Ca^2＋超载、释放内源性介质、激活再灌注损伤补救激酶（reperfusion injury salvage kinase,RISK）、抑制mPTP开放,从而保护心肌细胞。     

TFAR19促进小鼠肝线粒体膜通透性转运孔的开放

TFAR19基因 (TF 1cellapoptosisrelatedgene 19)是北京大学人类疾病基因中心从人白血病细胞株TF 1细胞中克隆到的凋亡相关新基因之一 (GenBank登记号AF0 1495 5 )。初步研究发现 ,该基因在细胞凋亡时高表达 ,并且表达产物具有抑制肿瘤细胞生长和促进凋亡作用。但是其确切的作用机制不明。线粒体膜完整性破坏所导致促凋亡因子 (如细胞色素c等因子 )的释放是细胞凋亡关键性的控制因素。线粒体膜通透性转运孔 (PTP) ,对线粒体膜完整性具有重要的调控作用。研究了重组人TFAR19蛋白在体外条件下 ,对线粒体PTP、跨膜电位 ,以及细胞色素c释放的影响。结果表明 ,TFAR19蛋白使分离的小鼠肝线粒体PTP开放、线粒体跨膜电位下降 ,以及细胞色素c释放。TFAR19对线粒体的上述作用是通过促进PTP开放起作用的。实验结果提示 ,TFAR19对线粒体凋亡信号有正反馈放大作用 ,并进一步揭示了TFAR19促进细胞凋亡的机制     

线粒体与细胞调亡

线粒体不仅是动物细胞内的主要产能中心,有理 在细胞凋亡中还承担着主开关的角色。作为细胞生死开关的线粒体,其通透性转变孔的开放,可让细胞色素c、凋记发诱导因子,Ca^2 以及膜间隙中的胱冬肽酶原（procaspase)等凋亡因子被释放到细胞质中,它们或激活凋亡蛋白家族的主要成员－－胱冬肽酶（caspase）,或独立地破坏核染色质,或作用于其它Ca^2 －依赖性蛋白,从而使细胞的整体结构破坏、功能紊乱、最终变成泡状凋亡小体而凋亡。弄清线粒体在细胞凋亡中的调控机制,不仅具有重要的理论意义,而且在开发研制以线粒体为靶目标的各种药物以治疗癌症、老年性痴呆和帕金森综合症等方面也具有重要的实践价值。本文谨以对线粒体及细胞凋亡的最新研究进展为基础,对线粒体在决定细胞生列中所起的关键作用做一综述。     

以线粒体通透性转换孔为靶点的脓毒症器官功能保护研究进展

脓毒症是由宿主对感染的反应失调引起的危及生命的器官功能障碍.对于脓毒症的治疗主要是抗感染、抗休克、维持机体组织器官灌注等.但近年来,在对脓毒症诱导的组织器官功能障碍的研究中发现,脓毒症时出现多器官功能障碍的原因不仅在于组织器官的缺血缺氧,而且与线粒体通透性转换孔(mitochondrial permeability t...     

线粒体通透性增高过程中新的参与者

 应激时,线粒体通透性增高可导致位于线粒体膜间的致凋亡因子释放入细胞质,胱天蛋白酶 (caspase)激活,以及细胞死亡.但线粒体通透性增高的确切机制尚不清楚.许多研究表明,线粒体通透性增高过程需要Bcl-2家族蛋白中促凋亡Bax亚家族蛋白,主要是Bax和Bak的激活;该家族中其它蛋白可对Bax和Bak进行调节.但最近的研究表明,其它非Bcl-2家庭蛋白的蛋白质包括抑制因子和激活因子,也可对线粒体通透性增高过程进行调节.此外,应激时线粒体脂质重新分布,对于线粒体膜通透性增高过程也起重要作用.     

线粒体PT孔与细胞凋亡

线粒体在细胞凋亡中有着重要作用,而线粒体PT孔开放是线粒体因素导致细胞凋亡的关键。促凋亡因素通过诱导PT孔的形成,导致线粒体膜电位丧失,使细胞色素C和凋亡诱导因子等释放进入细胞浆,启动细胞凋亡程序,诱导细胞凋亡。本文就目前线粒体PT孔的结构、可能的凋控机制及其在细胞凋亡中作用的研究进展进行综述。     

Modulation of mitochondrial permeability transition pore affects multidrug resistance in human hepatocellular carcinoma cells

Multidrug resistance (MDR) is a critical problem in the chemotherapy of cancers. Human hepatocellular carcinoma (HCC) responds poorly to chemotherapy owing to its potent MDR. Chemotherapeutic drugs primarily act by inducing apoptosis of cancer cells, and defects in apoptosis may result in MDR. Mitochondrial permeability transition (mPT) is implicated as an important event in the control of cell death or survival and mPT represents a target for the development of cytotoxic drugs. This study aimed to investigate the effects of selective opener (Atractyloside glycoside, ATR) and inhibitor (Cyclosporine A, CsA) of mitochondrial permeability transition pore (mPTP) on a CDDP-resistant HCC cell line (SK-Hep1 cells). In this study, a stable MDR phenotype characterization of SK-Hep1 cell line (SK-Hep1/CDDP cells) was established and used to investigate the role of mPTP in MDR. Results suggested that ATR accelerated the decrease of mitochondrial membrane potential (ΔΨm), reduced the Bax activity, and increased the apoptosis of SK-Hep1/CDDP cells; while CsA inhibited mPTP opening, reduced and delayed the decline of mitochondrial membrane potential, and increased the Bax activity, leading to increased tolerance of SK-Hep1/CDDP cells to apoptosis induction. However, mPTP activity had no effect on the expression of MDR1 in cells,meanwhile the P-gp translocation to mitochondria was increased, and functionally activated. In conclusion, selective modulation of mPTP can affect MDR in human HCC cells. Therefore, activation of mPTP may provide a new strategy to sensitize cancer cells to chemotherapeutic drugs and to reverse the MDR in cancer cells.     

The mitochondrial permeability transition pore: an evolving concept critical for cell life and death

In this review, we summarize current knowledge of perhaps one of the most intriguing phenomena in cell biology: the mitochondrial permeability transition pore (mPTP). This phenomenon, which was initially observed as a sudden loss of inner mitochondrial membrane impermeability caused by excessive calcium, has been studied for almost 50 years, and still no definitive answer has been provided regarding its mechanisms. From its initial consideration as an in vitro artifact to the current notion that the mPTP is a phenomenon with physiological and pathological implications, a long road has been travelled. We here summarize the role of mitochondria in cytosolic calcium control and the evolving concepts regarding the mitochondrial permeability transition (mPT) and the mPTP. We show how the evolving mPTP models and mechanisms, which involve many proposed mitochondrial protein components, have arisen from methodological advances and more complex biological models. We describe how scientific progress and methodological advances have allowed milestone discoveries on mPTP regulation and composition and its recognition as a valid target for drug development and a critical component of mitochondrial biology.     

随年龄增长心肌线粒体PT孔开放改变及其机制

目的:研究心功能自然衰退过程中线拉体通透性转换孔(MPTP)开放改变规律及其相关机制.方法:检测不同月龄(3、6、9、12月龄)SD大鼠左室心功能;分离各月龄大鼠心肌线粒体,检测MPTP开放改变、线粒体Mn-SOD活性.结果:9月龄和12月龄大鼠心功能同3月龄大鼠相比均出现明显减退,表现为左室收缩压LVSP减小(P<0...     

Mitochondrial Ca2+ transport and permeability transition pore opening and mitochondrial energetic status

The relationship between mitochondrial Ca2+ transport and permeability transition pore (PTP) opening as well as the effects of mitochondrial energetic status on mitochondrial Ca2+ transport and PTP opening were studied. The results showed that the calcium-induced calcium release from mitochondria (mCICR) induced PTP opening. Inhibitors for electron transport of respiratory chain inhibited mCICR and PTP opening. Partial recovery of electron transport in respiratory chain resulted in partial recovery of mCICR and PTP opening. mCICR and PTP opening were also inhibited by CCCP which eliminated transmembrane proton gradient. The results indicated that mitochondrial Ca2+ transport and PTP opening are largely dependent on electron transport and energy coupling.     

Mitochondrial Ca2+ transport and permeability transition pore opening and mitochondrial energetic status

The relationship between mitochondrial Ca2 transport and permeability transition pore (PTP) opening as well as the effects of mitochondrial energetic status on mitochondrial Ca2 transport and PTP opening were studied. The results showed that the calcium-induced calcium release from mitochondria (mClCR) induced PTP opening. Inhibitors for electron transport of respiratory chain inhibited mClCR and PTP opening. Partial recovery of electron transport in respiratory chain resulted in partial recovery of mClCR and PTP opening. mClCR and PTP opening were also inhibited by CCCP which eliminated transmembrane proton gradient. The results indicated that mitochondrial Ca2 transport and PTP opening are largely dependent on electron transport and energy coupling.     

Participation of the mitochondrial permeability transition pore in nitric oxide-induced plant cell death

In the present study, we investigated the involvement of the mitochondrial permeability transition pore (PTP) in nitric oxide (NO)-induced plant cell death. NO donors such as sodium nitroprusside (SNP) and S-nitroso-N-acetylpenicillamine inhibited growth and caused death in suspension-cultured cells of Citrus sinensis. Cells treated with SNP showed chromatin condensation and fragmentation, characteristic of apoptosis. SNP caused loss of the mitochondrial membrane electrical potential, which was prevented by cyclosporin A (CsA), a specific inhibitor of PTP formation. CsA also prevented the nuclear apoptosis and subsequent Citrus cell death induced by NO. These findings indicate that mitochondrial PTP formation is involved in the signaling pathway by which NO induces apoptosis in cultured Citrus cells.     

Discovery of a new mitochondria permeability transition pore (mPTP) inhibitor based on gallic acid

Pharmacological interventions targeting mitochondria present several barriers for a complete efficacy. Therefore, a new mitochondriotropic antioxidant (AntiOxBEN₃) based on the dietary antioxidant gallic acid was developed. AntiOxBEN₃ accumulated several thousand-fold inside isolated rat liver mitochondria, without causing disruption of the oxidative phosphorylation apparatus, as seen by the unchanged respiratory control ratio, phosphorylation efficiency, and transmembrane electric potential. AntiOxBEN₃ showed also limited toxicity on human hepatocarcinoma cells. Moreover, AntiOxBEN₃ presented robust iron-chelation and antioxidant properties in both isolated liver mitochondria and cultured rat and human cell lines. Along with its low toxicity profile and high antioxidant activity, AntiOxBEN₃ strongly inhibited the calcium-dependent mitochondrial permeability transition pore (mPTP) opening. From our data, AntiOxBEN₃ can be considered as a lead compound for the development of a new class of mPTP inhibitors and be used as mPTP de-sensitiser for basic research or clinical applications or emerge as a therapeutic application in mitochondria dysfunction-related disorders.     

谷氨酰胺对心肌细胞线粒体膜PTP开放干预作用的实验研究

目的:探讨谷氨酰胺(Gln)对过度训练状态下心肌线粒体膜通透性转换孔(PTP)开放的干预作用及其可能机制。方法:30只SD大鼠随机分为3组(n=10):对照组(CG组)、过度训练组(OG组)和补充Gln+过度训练组(GOG组)。采用分光光度法检测大鼠心肌线粒体PTP开放程度,电化学法检测心肌丙二醛(MDA)、还原型谷胱苷肽(GSH)含量和磷脂酶A2(PLA2)活性。结果:OG组与GOG组比较,吸光度(A0)显著下降(P<0.05),吸光度变化(△A)值显著降低(P<0.05);荧光剂罗丹明123(Rh123)的荧光强度(F0)显著增强(P<0.05),Rh123荧光强度变化(△F)值明显降低(P<0.05)。与GOG组比较,线粒体GSH含量显著降低(P<0.05),PLA2活性显著增加(P<0.05);MDA含量显著升高(P<0.05)。结论:过度训练可导致心肌细胞线粒体PTP开放增加,过度训练状态下线粒体活性氧生成增多,PLA2活性增加及GSH的含量下降,补充外源性的Gln对这些变化有显著的干预作用。