线粒体与细胞凋亡的研究进展

细胞凋亡 (apoptosis)是一种普遍的生理现象 ,是指在一定的生理或病理条件下 ,按程序进行的细胞死亡过程 ,亦称程序性细胞死亡 (programmedcelldeath ,PCD) ,对于维持多细胞机体的完整性和自身稳态具有重要作用。线粒体是所有真核细胞内腺苷三磷酸(ATP)产生中心 ,对维持细胞能量代谢和正常功能活动起重要作用。新近研究发现 ,线粒体内包含一些与细胞凋亡有密切关系的物质 ,如细胞色素C(CytC、凋亡诱导因子 (AIF)、Ca2 + 和活性氧自由基 (ROS)等。在凋亡信号的刺激下 ,线粒体膜通透性增加 ,由此产生一系列关键性变化 ,包括CytC的释放…     

细胞色素C、线粒体与凋亡

线粒体细胞色素C释放在细胞凋亡过程中起重要作用。细胞色素C释放到胞质后可引发caspase活化级联 ,导致细胞死亡。细胞色素C的释放是线粒体外膜通透性增高的结果。Bcl 2蛋白家族具有调控细胞色素C释放的功能。除细胞色素C外 ,线粒体膜间隙的凋亡诱导因子 (AIF)在凋亡过程中也释放至胞质 ,这两种途径充分保证了细胞死亡程序的有效执行。轻中度暂时性脑缺血后细胞色素C释放至胞质 ,早于DNA片段化     

利拉鲁肽基于线粒体-细胞色素C介导心肌细胞凋亡

目的 探讨利拉鲁肽在缺氧/复氧（hypoxia/reoxygenation,H/R）处理的心肌细胞中的作用及其潜在机制。方法 建立大鼠心肌细胞株（H9C2） H/R模型以模拟体外心肌缺血再灌注（ischemia/reperfusion,I/R）损伤。用四甲基偶氮唑盐[3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide,MTT]和Annexin V FITC-PI染色法测定细胞增殖和凋亡。检测细胞氧化应激产物和线粒体功能。用蛋白印迹法（Western blotting）检测细胞凋亡蛋白表达和细胞色素C的释放。结果 利拉鲁肽保护H9C2细胞免受H/R诱导的损伤,因其可减弱H/R对H9C2细胞活力、细胞凋亡和活性氧（reactive oxygen species,ROS）产生的影响（P<0.05）。利拉鲁肽可保护线粒体功能并防止H/R损伤后H9C2细胞中细胞色素C的释放（P<0.05）。结论 阐明了利拉鲁肽在治疗I/R相关心肌损伤中的潜在心血管保护作用。     

辛伐他汀通过线粒体途径诱导K562细胞凋亡

目的观察辛伐他汀诱导K562细胞凋亡不同时间的膜电位(Δψm),caspase-3、9和细胞色素C的改变,以推测其凋亡通路。方法采用浓度为20μmol.L-1的辛伐他汀处理K562细胞24、48、72 h,采用流式细胞技术检测细胞凋亡率和线粒体膜电位,分光光度法检测caspase-3、9蛋白活性,免疫组织化学法检测细胞色素C蛋白。结果浓度为20μmol.L-1辛伐他汀作用K562细胞24、48、72 h后,凋亡率分别为(6.1±0.35)%、(14.15±0.42)%(、30.70±0.65)%,随着凋亡率增加线粒体膜电位降低分别为(39.6±4.80)%,(24.4±2.45)%,(6.0±1.62)%;caspase-3、9蛋白活性与对照组相比上调,细胞浆内细胞色素C升高。结论辛伐他汀诱导K562细胞凋亡时线粒体膜电位下降,caspase-3、9活性增高和细胞色素C释放,推测辛伐他汀诱导K562细胞的凋亡可能经过线粒体凋亡途径。     

大鼠脑出血模型出血灶周围神经细胞凋亡的研究

目的 研究大鼠自体动脉血脑出血模型出血灶周围神经细胞的凋亡.方法 应用立体定向技术,用大鼠自体尾动脉不抗凝动脉血建立自体动脉血脑出血动物模型,动态观察其行为学改变以及出血灶周神经细胞形态结构,并用免疫组化的方法检测出血灶周神经细胞内Bcl-2和细胞色素C的表达.结果 大鼠脑出血后不同观察时间点肢体对称试验评分、Longa评分和Berder son评分与对照组比较,差异具有统计学意义(P＜0.05).脑出血周围神经细胞的形态结构较对照组有显著差别;出血灶周神经细胞内细胞色素C的表达明显高于对照组,Bcl-2的表达明显低于假手术组.结论 大鼠自体动脉血脑出血动物模型是比较理想的实验性脑出血模型,在出血灶周围存在大量神经细胞的凋亡.     

力达霉素经线粒体依赖通路介导细胞凋亡

力达霉素(lidamycin，LDM)是具有我国自主知识产权的烯二炔类抗生素，具有较强的抗肿瘤活性，其抗肿瘤机制有待深入阐明。本研究主要利用人肝癌BEL-7402和乳腺癌MCF-7细胞研究了力达霉素对线粒体凋亡通路相关因子的影响。通过MTT法检测不同剂量力达霉素对肿瘤细胞增殖毒性；蛋白印迹技术检测线粒体和细胞质细胞色素c、总Bax，Bcl-2及p53表达；RT-PCR检测p53和bax mRNA表达等。结果发现，LDM能迅速激活肿瘤细胞线粒体凋亡通路，在2 h内就引起线粒体中细胞色素c释放至细胞质中。同时Bcl-2家族成员中促凋亡成员Bax持续增加，而抗凋亡成员Bcl-2先增加后减少。p53蛋白在6 h显著增加。Bax蛋白表达升高与其mRNA转录水平升高有关，p53蛋白表达升高与转录后水平有关。 Caspase抑制剂可以部分抑制LDM的增殖毒性。因此LDM可以通过激活细胞色素c启动的线粒体凋亡通路发挥抗肿瘤作用。     

哈巴苷对急性脑缺血及线粒体介导的Caspase依赖性细胞凋亡信号通路的影响

目的探讨哈巴苷对急性脑缺血及线粒体介导的caspase依赖性细胞凋亡通路的影响。方法采用MCAO法建立小鼠急性脑缺血模型,造模后各组立即尾静脉注射(iv)哈巴苷(4、8、12 mg·kg~(-1))、依达拉奉(3.2 mg·kg~(-1)),模型组及假手术组同法给予等量生理盐水,0.1 mL·(10 g)~(-1)。造模6h后,观测小鼠神经功能评分、脑梗死体积及脑组织形态病理学变化;采用Western blot法测定各组脑组织线粒体中Cyt C及胞质中pro-caspase-3的含量。结果造模6 h后,哈巴苷各剂量组均能明显降低因缺血而增加的小鼠神经功能评分及脑梗死体积(P<0.01,P<0.05);均能不同程度减轻脑组织神经元核固缩、核溶解程度,改善脑组织因缺血造成的病理损伤;能明显上调脑组织线粒体中Cyt C及胞质中pro-caspase-3的表达。结论哈巴苷对急性脑缺血具有保护作用,其保护作用可能与抑制线粒体介导的caspase依赖性细胞凋亡信号通路相关。     

细胞凋亡的线粒体途径机制研究

细胞凋亡是细胞有机体为调控机体发育,维护内环境稳定而主动采取的、由基因决定的、自动结束细胞生命的过程。线粒体在细胞凋亡中发挥着重要的作用。线粒体通透性转变孔（MPTP）开放导致线粒体内释放出包括细胞色素C（cyt-c、Smac／DIABLO和AIF等线粒体凋亡因子在内的各种蛋白,从而启动细胞凋亡程序引起细胞凋亡。Cyt—c释放到细胞浆后,Cyt—c、dATP、Aapf-1和procaspase-9组成聚合体,称为凋亡体。凋亡体可激活其下游的procaspase-3,从而导致细胞死亡。在凋亡诱导因素的刺激下,AIF可从线粒体转位进入胞核,直接引起染色质浓缩及DNA的断裂。AIF的转位足以介导体外细胞凋亡的发生。     

白藜芦醇诱导鼻咽癌细胞CNE-2Z凋亡的线粒体机制(英文)

目的　探讨白藜芦醇诱导鼻咽癌细胞CNE 2Z凋亡的线粒体机制。方法　用 10 0 μmol·L- 1白藜芦醇分别处理细胞 0 (对照 ) ,2 ,4 ,8,12 ,2 4h和分别用 0 ,2 5 ,5 0 ,10 0 ,2 0 0 μmol·L- 1白藜芦醇处理CNE 2Z细胞 8h ,采用Western印迹分析Bcl 2 ,Bax和胞浆中细胞色素C(CytC)的蛋白水平变化 ,罗丹明 12 3荧光染色后经流式细胞术检测线粒体膜电位(△ψm)变化 ,比色法测定半胱天冬酶 9的活性改变。结果　① 10 0 μmol·L- 1白藜芦醇处理细胞不同时间 ,Bcl 2蛋白表达和△ψm 减少、胞浆中的CytC和半胱天冬酶 9活性增高均呈时间依赖性 (P <0 .0 1) ,但Bax蛋白表达无改变。除Bax以外的其他指标均自白藜芦醇处理 2h即有变化。Bcl 2的蛋白表达受抑和△ψm 的丧失在 4～ 8h间最明显 (与对照组比较P <0 .0 1) ,在 2 4h时已无意义。胞浆中CytC水平和半胱天冬酶 9活性在 8h达高峰 (分别为 0h的 3.0 ,5 .4倍 ) ,2 4h时仍明显高于对照组(P <0 .0 1)。②细胞经不同浓度的白藜芦醇处理 8h后 ,Bcl 2的蛋白表达受抑、△ψm 的丧失、CytC的释放和半胱天冬酶 9活性的升高均具有剂量依赖性 (P <0 .0 1) ,但Bax的蛋白表达未受影响。结论白藜芦醇可能经一个线粒体 /半胱天冬酶 9的特定途径诱导CNE 2Z细胞凋亡 ,但此凋亡?     

Curcumin protects mitochondria from oxidative damage and attenuates apoptosis in cortical neurons

AIM: To investigate the effect of curcumin on tert-butyl hydroperoxide (t-BHP)-induced oxidative damage in rat cortical neurons and to explore the possible mechanism. METHODS: Primary cultured rat cortical neurons wereperformed in vitro and cell viability was measured by MTT assay. DNA fragmentation was used to evaluate cellapoptosis. Intracellular reactive oxygen species (ROS) and mitochondrial membrane potential (Aψm) was determined by flow cytometric assay. Cellular glutathione (GSH) content was measured by spectrophotometer.‘ Bcl-2family proteins, cytochrome c, cleaved caspase-3, and poly (ADP-ribose) polymerase (PARP) were detected byWestern blot. RESULTS: Exposure of tBHP 100 μmol/L to neurons for 60 rain resulted in △ψm loss and cyto-chrome c release from mitochondria and subsequent activation of caspase-3 and PARP cleavation, and cell apoptosis.After removal of tBHP and then further treatment with curcumin (2.5-20 μmol/L) for 18 h, curcumin abrogated △ψm loss and cytochrome c release, blocked activation of caspase 3, and altered the expression of Bcl-2 family.Further curcumin treatment also prevented cellular GSH and decreased intracellular ROS generation markedly.Curcumin eventually attenuated tBHP-induced apoptosis in cortical neurons. CONCLUSION: Curcumin mayattenuate oxidative damages in cortical neurons by reducing intracellular production of ROS and protecting mito-chondria from oxidative damage.     

斑秃患者外周血淋巴细胞凋亡的研究

目的:研究斑秃患者外周血淋巴细胞凋亡调控基因表达蛋白CD95和bcl2的表达,以探讨其在斑秃发病中所起的作用。方法:采用流式细胞仪技术检测CD95和bcl2在外周血淋巴细胞上的表达。结果:斑秃患者外周血淋巴细胞CD95表达率较正常对照组显著增高(P<0.01),而bcl2的表达率与正常对照组相比差异无显著性(P>0.05)。结论:CD95在斑秃患者外周血淋巴细胞上的表达增高,可诱导毛囊细胞快速凋亡,使斑秃在短时间内发生并促其发展。     

The new research on mitochondria

Apoptosis,also known as programmed cell death,is the removal of damaged body organizations,aging or redundant cells in a suicide,has to maintain the health of the body,the normal development of the nervous system,the immune system to maintain the normal function of such areas is of great significance.The morphological characteristics of apoptosis are the cytoplasm concentrated,condensed nuclear chromatin,DNA fragments of a large-scale,the cell membrane invagination and foam formation of apoptotic bodies.There are two classic apoptosis ways which are generally accepted by majority of the scholars currently:Mitochondrial pathway and Death receptor pathway.Mitochondrial membrane is a two-tier structure surrounded the cystic,between the external cavity and internal cavity which is called the Room,surrounded by the internal cavity known as the mitochondria internal room or mitochondrial matrix.Mitochondria with the functions of control cell survival and death:mitochondria play an important role in physiological such as oxidative phosphorylation,electronic transfer,storage Ca2+,energy metabolism,anti-oxidation activity and so on,to provide the basic energy to the various activities of cell life.Study found that mitochondria contain some of the material is closely related to apoptosis,such as Cyt-C,Smac/Diablo,AIF,Ca2+,ROS and so on.In the signal to stimulate apoptosis,mitochondrial membrane permeability,resulting in a series of key changes,including Cyt-C,Smac/Diablo release,decline of the mitochondrial membrane potential(ψm),the state of the redox within cells,the intervention of Bcl gene family and so on.Different signal transduction ultimately focuses on the mitochondria to activate or inhibit these incidents,then the corresponding signal transduction to control apoptosis.Therefore,the mitochondria in the incidence of apoptosis play an important role.In recent years,the study confirmed that apoptosis imbalance can cause a variety of human diseases.And there is a close relationship among apoptosis and tumor occurrence,development and dissipated.The resistance to apoptosis caused by the disorders of tumor cell apoptosis control is one of the important reasons which cause tumor.Therefore,to prove the mechanisms of apoptosis in mitochondrial pathway and use these mechanisms to prevent and treat cancer is of great importance.     

GSK3 promotes arsenite-induced apoptosis via facilitation of mitochondria disruption

Arsenic is an environmental toxicant that recently has been shown to have anticancer activity against a number of types of cancer cells by inducing apoptosis. Glycogen synthase kinase-3 (GSK3), a serine/threonine kinase, is an important pro-apoptotic signaling enzyme. Although GSK3 has been shown to promote apoptosis caused by a wide variety of insults, a role for GSK3 in arsenic-induced apoptosis has not yet been identified. Investigation of the involvement of GSK3 in arsenite-induced apoptosis demonstrated that arsenite induced apoptosis in SH-SY5Y human neuroblastoma cells, activating the executioner caspase-3 which caused cleavage of poly-ADP ribose-polymerase (PARP). Two selective GSK3 inhibitors, lithium and SB216763, attenuated caspase-3 activation and PARP cleavage induced by arsenite treatment indicating that GSK3 contributed to arsenite-induced apoptosis. Apoptotic signaling following exposure to arsenite involved cytochrome C release from mitochondria, and this was reduced by inhibition of GSK3 indicating that GSK3 promotes arsenite-induced apoptotic signaling upstream of mitochondrial disruption. Moreover, arsenite induced the translocation of Bax and p53 to the mitochondria and the activation-associated oligomerization of Bax, and these crucial events were reduced by inhibition of GSK3, indicating that GSK3 promotes arsenite-induced apoptosis by facilitating signals leading to mitochondrial apoptotic events. Taken together, the findings from this study reveal that GSK3 promotes arsenite-induced apoptosis by facilitating signaling leading to disruption of mitochondria.     

喜树碱诱导人白血病HL-60细胞凋亡过程中端粒酶的调控(英文)

目的:研究在喜树碱诱导人白血病HL—60细胞凋亡过程中端粒酶的调节变化规律.方法:用MTT法测定药物对细胞存活率的影响;用琼脂糖电泳及流式细胞术检测和定量凋亡的发生;用以PCR为基础的TRAP法测定端粒酶活力;逆转录PCR检测凋亡过程中bcl-2及端粒酶亚基hTR、hEST2/hTERT和TLPl/TPl的基因表达水平的变化.结果:端粒酶活力伴随喜树碱诱导HL-60细胞凋亡的发生而逐渐降低,在此过程中端粒酶各亚基的mRNA水平无可见性变化,而bcl-2的基因表达水平则相应下调.结论:端粒酶活力的下调和喜树碱诱导的HL-60凋亡密切相关,端粒酶活力的阻断并非发生在其亚基基因转录水平,bcl-2对端粒酶活力的调节也不是通过影响端粒酶亚基的转录水平来实现的.     

Staurosporine诱导人乳腺癌MCF7/GFP-Bax非受体依赖途径的细胞凋亡对Bax自胞浆至线粒体转移定位的影响

以建立的人乳腺癌MCF7 细胞GFP-Bax稳定表达细胞株(MCF7/GFP-PBax)，观察staurosporine(STS)诱导的非受体途径的细胞凋亡对Bax自胞浆转移定位于线粒体的影响。荧光显微镜观察凋亡细胞Bax从细胞浆至线粒体转移和细胞核染色体断裂，检测STS诱导细胞凋亡的量效和时效关系。免疫荧光法观察GFP-Bax从细胞浆转移至线粒体与定位、细胞色素c(cytochrome c，Cyt-c)释放和Annexin V染色。MTT法测定STS的细胞毒作用，TMRE观测对细胞线粒体膜电位(ΔΨm)与功能的影响。Western blotting方法分析STS诱导的细胞凋亡的信号转导途径和作用机制。结果STS可明显促进Bax从细胞浆转移至线粒体与定位、细胞色素c释放，Western blotting显示JNK特异抑制剂SP600125可抑制STS诱导的细胞pJNK表达，表明STS作用机制与激活JNK信号通路相关。     

Cisplatin-induced apoptosis is enhanced by hypoxia and by inhibition of mitochondria in renal collecting duct cells.

Cisplatin is a widely used chemotherapeutic agent. Here we show that cisplatin induces apoptosis in renal collecting duct-derived cells (MDCK-C7 cells, resembling principal cells) in a dose-dependent manner. Additionally, we studied the role of mitochondria in this process by inhibition of the mitochondrial respiratory chain, the F1F(o)-ATP synthase or by uncoupling. The role of intra- and extracellular pH in apoptosis induction was investigated. Activation of caspase-3 and DNA ladder formation were used to monitor the apoptotic response. When cells were incubated with inhibitors of the mitochondrial respiratory chain or an inhibitor of the ATP-synthase, cisplatin-induced apoptosis was markedly enhanced. Mitochondrial blockade led to enhanced production of lactic acid. Also, anoxia potentiated the cisplatin-induced caspase-3 activation. Neither intra- nor extracellular pH had an influence on caspase-3 activation at low cisplatin concentrations. Acidic conditions (pH 6.8) potentiated the caspase-3 activation when high (100 microM) cisplatin concentrations were used. We demonstrate that intact mitochondria are important to prevent cisplatin-induced apoptosis in MDCK-C7 cells and that acidic conditions can aggravate the toxic effects of cisplatin.     

线粒体靶向的肿瘤治疗研究进展

线粒体控制着细胞凋亡的激活系统,肿瘤细胞的多种特征,包括无尽的增殖能力,对抑制生长信号的不敏感,受损的细胞凋亡机制等都和线粒体的机能丧失有关。该文综述了近年来线粒体靶向的抗肿瘤研究进展,重点介绍新发现的针对线粒体为靶点的药物及其作用方式与潜在临床应用价值,展示人类恶性肿瘤治疗的前景。     

黄酮类化合物对心肌细胞凋亡的作用

目的观察黄酮类化合物包括高良姜素(G)、山萘酚(K)、芹菜素(A)、桑色素(M)、槲皮素(Q)、杨梅素(MY)对H2O2诱导的心肌细胞凋亡的作用,并探讨其可能的作用机制。方法培养新生Wistar乳鼠心肌细胞,随机分为正常组、模型组、药物组。分别用荧光染色法、琼脂糖凝胶电泳法、Western blot法观察心肌细胞凋亡情况及蛋白表达的变化。结果100μmol·L-1H2O2孵育心肌细胞16 h能明显诱导心肌细胞凋亡(P<0.01),黄酮类化合物能降低细胞凋亡率,抑制DNA损伤,上调bcl-2、bcl-xl,下调bax蛋白表达。结论黄酮类化合物能抗氧化,从而抑制心肌细胞凋亡,主要是通过线粒体死亡途径调节bcl-2家族蛋白表达来发挥作用的。