内源性蛋白酶对宰后肌肉嫩化机制研究进展

 肌肉的宰后嫩化机制一直是国内外肉品科学研究的一个热点,目前认为肌肉宰后嫩度的改善主要归因于内源性水解酶作用所引起的肌原纤维蛋白的有限降解。钙激活酶虽然是宰后嫩度改善的一个主要贡献者,但不是惟一的,而是多种内源酶类协同作用的结果。本文综述了溶酶体组织蛋白酶、蛋白酶体、钙激活酶在宰后嫩化过程的作用;介绍了细胞凋亡的特点、通路及凋亡酶,并对细胞凋亡酶对宰后嫩度改善的潜在贡献进行了讨论;最后分析了细胞凋亡酶和钙激活酶在宰后成熟过程中的交互作用及共同贡献于肌肉宰后嫩化的可能性。     

细胞凋亡与细胞色素C

细胞凋亡是动植物最基本的生命活动,是一个有一系列酶参与的,并且由基因控制的主动的、高度有序的死亡过程。线粒体除了为细胞提供能量外,在细胞凋亡中也起着中心调控作用。研究发现,线粒体释放的细胞色素C是细胞凋亡过程的关键因素,已是近些年研究的热点。本文就细胞色素C从线粒体释放的机制及在凋亡中的作用进行综述。     

钙蛋白酶系统对肌肉嫩化的影响

宰后肌肉的嫩化主要由钙蛋白酶系统降解肌原纤维引起的,综述了钙蛋白酶系统的结构,对肌肉嫩化的作用机理和它的活化过程。     

电离辐射与线粒体DNA的变异

线粒体DNA(mitchondrial DNA,mtDNA)是裸露的双链环状DNA分子结构．缺少组蛋白外衣的保护,复制快速．缺乏校读功能以及缺乏有效的DNA修复系统,极易发生突变。而电离辐射作为一种致DNA损伤的因素．现已发现它可以诱导线粒体DNA的突变,其中包括4977bp的缺失、点突变、插入突变等。在凋亡调节诸多通路中,线粒体在电离辐射诱导的细胞凋亡通路中起中心调控作用。各种死亡信号通过Bcl-2家族蛋白或直接诱导线粒体膜的通透性的改变、细胞色素C释放和caspases激活,最终引起细胞凋亡。线粒体膜的通透性改变的机制．目前还未完全明了。本文就电离辐射与线粒体DNA的变异以及线粒体DNA变异与凋亡的关系作一简要综述。     

家蚕细胞色素C基因的克隆及其蛋白在家蚕凋亡细胞中的释放

 【目的】细胞色素C是线粒体凋亡路径上的关键因子,通过家蚕细胞色素C基因的克隆和其蛋白在家蚕凋亡细胞中的释放研究,为家蚕细胞凋亡的研究奠定基础。【方法】利用生物信息学和分子生物学方法克隆家蚕细胞色素C基因,通过紫外线照射诱导家蚕细胞凋亡,应用流式细胞仪和Western-blotting检测家蚕凋亡细胞的线粒体膜电位变化和细胞色素C的释放。【结果】在家蚕中克隆了一个含有细胞色素C保守结构域的同源基因,该基因cDNA长570 bp,有3个外显子,开放阅读框（open reading frame,ORF）长324 bp,编码108个氨基酸,存在参与细胞凋亡时与下游凋亡蛋白酶激活因子（Apaf－1）相互作用的保守关键位点;紫外线诱导家蚕胚胎细胞BmE-SWU1凋亡后12 h,细胞线粒体跨膜电位明显下降,同时检测到了细胞色素C由线粒体向细胞质释放。【结论】在家蚕细胞凋亡中可能存在细胞色素C由线粒体释放的路径。     

血管紧张素Ⅱ对牛肉品质的影响

取屠宰后5头杂交牛(鲁西黄牛×西门塔尔)的背最长肌,置于2~4℃贮藏7 d,注射样品重10%的血管紧张素Ⅱ,观察在成熟过程中血管紧张素Ⅱ对牛肉剪切力、肌原纤维小片化指数(MFI)、凋亡酶活力,以及肌纤维直径变化的影响。结果显示:贮藏期间对照组和处理组样品剪切力均显著下降(P<0.05),MFI值逐渐增大,caspase-3、6、9酶活力宰后迅速激活随后其活力逐渐下降;血管紧张素Ⅱ注射处理显著降低了牛肉的剪切力值和肌纤维直径(P<0.05),而对MFI值及caspase-3、6、9活力无显著影响。因此,caspase-3、6、9对牛肉嫩度的影响可能仅发生在宰后初期极短的一段时间内;肌细胞宰后生理和活体状况下不太一致,血管紧张素Ⅱ注射降低了牛肉剪切力可能由于它能显著防止肌纤维的收缩。     

低温诱导淡水白鲳细胞凋亡的研究

为探讨10℃低温导致淡水白鲳尾鳍细胞系CBT死亡的原因及其可能机制.采用CCK-8试剂盒法检测细胞存活率,荧光染料H2DCFDA染色测定细胞内活性氧(ROS)含量,亚二倍体峰和Tunel分析检测细胞凋亡,琼脂糖凝胶电泳进行DNA片段分析,Hoechst33258染色观察凋亡细胞核形态变化,比色法测定乳酸脱氢酶(LDH)释放率.结果表明随CBT细胞受10℃低温作用时间的延长,细胞存活率显著下降,细胞内ROS含量和LDH释放率显著增加(P＜0.01).低温处理3 d的CBT细胞经流式细胞仪检测出现亚二倍体峰,凋亡率为13%.再经32℃复温12 h后,琼脂糖凝胶电泳显示DNA凋亡梯带;Tunel检测,低温处理3、4和5 d的CBT细胞分别有23.49%,27.72%和35.10%发生凋亡.低温处理5 d的CBT细胞核分裂成多个小核,呈现出典型的凋亡小体.因此10℃低温能诱导CBT细胞凋亡.ROS参与了10℃低温致CBT细胞损伤及凋亡过程.     

犬细小病毒通过促进ROS产生/线粒体损伤诱导宿主细胞的凋亡

为探讨犬细小病毒(CPV)引起宿主细胞凋亡的分子机制,用CPV感染MDCK细胞,感染后通过台盼蓝染色法检测不同时间的细胞存活率,用Annexin V-FITC/PI法检测磷脂酰丝氨酸外翻情况,用试剂盒检测Caspase-3活性,分析CPV诱导细胞的凋亡。并通过流式细胞术检测细胞线粒体的损伤和ROS的产生探讨其分子机制。结果显示,随着病毒感染时间的不断延长,CPV可明显降低MDCK细胞存活率,促进磷脂酰丝氨酸外翻,增强Caspase-3活性,表明CPV可引起细胞凋亡。同时还发现CPV可明显提高细胞内ROS的水平,引起线粒体的损伤,这可能是CPV诱导细胞凋亡的重要分子基础。由此可见,CPV诱导MDCK细胞凋亡与CPV增加细胞ROS的产生和线粒体的损伤有关。     

细胞凋亡的信号通路

细胞凋亡是细胞程序性死亡的一种方式,与自噬和坏死有明显的区别。细胞凋亡的信号途径比较复杂,在凋亡诱导因子的刺激下经历不同的信号途径。本文就细胞凋亡的三条信号通路——线粒体途径、内质网途径和死亡受体途径做一综述,以便为人们进一步了解细胞凋亡发生的机制,从而对癌症及其他一些相关疾病的治疗奠定基础。     

毛竹竹秆基本组织发育过程中ATP酶的超微定位

利用电镜细胞化学技术埘毛竹[Phyllostachys edulis(Carr.)H De Lehaie]竹秆基本组织发育过程中的ATP酶进行细胞化学定位.竹秆基本组织细胞分化早期,细胞具有较高的ATP酶活性,细胞核、质膜、内质网、线粒体等细胞器的膜系统上都具有ATP酶活性.随着基本组织的分化和发育,长细胞液泡膜上的ATP酶活性增强,而短细胞液泡膜上 ATP酶活性相对较弱;至仞生肇发育后期,质膜上的ATP酶活性升高;次生壁发育期,短细胞质膜上的ATP酶活性显著增强,而长细胞的相对较弱;细胞核、质膜、线粒体、胞间连丝、内质网、质体膜、运输小泡膜以及细胞质内和胞间隙都具有ATP酶活性.在次生壁发育期液泡膜上已观察不到ATP酶反应物.长细胞质膜ATP酶活性从第4年开始降低,短细胞质膜的ATP酶活性始终很高,且无论生长期还是休眠期,短细胞一直保持旺盛的物质主动吸收和活跃的新陈代谢过程.同时短细胞内大量的运输小泡,具有的ATP酶活性,以及胞间连丝沉积有大量的ATP酶反应物,都表明短细胞与周围细胞间频繁、活跃的物质交流.短细胞不仅在物质运输起到重要作用,而且在竹秆继续成熟的过程中可能参与长细胞次生壁的形成.     

肉品的嫩化机制、方法及其影响因素

肉品嫩度是肉的主要食用品质之一,它是消费者评判肉质优劣的最常用指标.由肌肉转变为可食性肉,到最后的嫩化成熟是一个复杂过程.宰后肉类的后期成熟和嫩化处理,是改善肉品嫩度的关键环节.该文主要阐述了影响肉嫩度的因素(宰前和宰后因素)、嫩化方法以及嫩化机制(如:电刺激、盐类、酶法、有机酸、超声波、高压和吊挂拉伸等),为肉制品加工企业品质改良提供理论支持.     

钙蛋白酶Ⅰ(CAPN1)基因的研究进展

对钙蛋白酶Ⅰ(CAPN1)的结构、活性调节、同源性、作用机制以及生物学功能的研究现状进行了综述,并讨论了其应用前景。     

钙蛋白酶系统基因的研究进展

钙蛋白酶系统主要由钙蛋白酶（calpainⅠ,CAPN1和calpainⅡ,CAPN2）,钙蛋白酶抑制蛋白（calpastatin,CAST）及骨骼肌特异性钙蛋白酶（muscle specific calpain,p94,CAPN3）组成。钙蛋白酶是细胞质中主要的蛋白水解酶,在肌原纤维蛋白降解中起着重要的作用。肌肉增长和宰后嫩度的变化与蛋白质水解程度密切相关。因此,钙蛋白酶的活性会影响畜禽肌肉增长和肉的嫩度。对钙蛋白酶系统基因的结构、活性调节、作用机理及其对肉质的影响进行了综述,并讨论其应用前景。     

The pathophysiology of mitochondrial cell death

In the mitochondrial pathway of apoptosis, caspase activation is closely linked to mitochondrial outer membrane permeabilization (MOMP). Numerous pro-apoptotic signal-transducing molecules and pathological stimuli converge on mitochondria to induce MOMP. The local regulation and execution of MOMP involve proteins from the Bcl-2 family, mitochondrial lipids, proteins that regulate bioenergetic metabolite flux, and putative components of the permeability transition pore. MOMP is lethal because it results in the release of caspase-activating molecules and caspase-independent death effectors, metabolic failure in the mitochondria, or both. Drugs designed to suppress excessive MOMP may avoid pathological cell death, and the therapeutic induction of MOMP may restore apoptosis in cancer cells in which it is disabled. The general rules governing the pathophysiology of MOMP and controversial issues regarding its regulation are discussed.     

BAX and BAK regulation of endoplasmic reticulum Ca2+: a control point for apoptosis

BAX and BAK are "multidomain" proapoptotic proteins that initiate mitochondrial dysfunction but also localize to the endoplasmic reticulum (ER). Mouse embryonic fibroblasts deficient for BAX and BAK (DKO cells) were found to have a reduced resting concentration of calcium in the ER ([Ca2+]er) that results in decreased uptake of Ca2+ by mitochondria after Ca2+ release from the ER. Expression of SERCA (sarcoplasmic-endoplasmic reticulum Ca2+ adenosine triphosphatase) corrected [Ca2+]er and mitochondrial Ca2+ uptake in DKO cells, restoring apoptotic death in response to agents that release Ca2+ from intracellular stores (such as arachidonic acid, C2-ceramide, and oxidative stress). In contrast, targeting of BAX to mitochondria selectively restored apoptosis to "BH3-only" signals. A third set of stimuli, including many intrinsic signals, required both ER-released Ca2+ and the presence of mitochondrial BAX or BAK to fully restore apoptosis. Thus, BAX and BAK operate in both the ER and mitochondria as an essential gateway for selected apoptotic signals.     

Proapoptotic BAX and BAK: a requisite gateway to mitochondrial dysfunction and death

Multiple death signals influence mitochondria during apoptosis, yet the critical initiating event for mitochondrial dysfunction in vivo has been unclear. tBID, the caspase-activated form of a "BH3-domain-only" BCL-2 family member, triggers the homooligomerization of "multidomain" conserved proapoptotic family members BAK or BAX, resulting in the release of cytochrome c from mitochondria. We find that cells lacking both Bax and Bak, but not cells lacking only one of these components, are completely resistant to tBID-induced cytochrome c release and apoptosis. Moreover, doubly deficient cells are resistant to multiple apoptotic stimuli that act through disruption of mitochondrial function: staurosporine, ultraviolet radiation, growth factor deprivation, etoposide, and the endoplasmic reticulum stress stimuli thapsigargin and tunicamycin. Thus, activation of a "multidomain" proapoptotic member, BAX or BAK, appears to be an essential gateway to mitochondrial dysfunction required for cell death in response to diverse stimuli.     

Protein kinase C beta and prolyl isomerase 1 regulate mitochondrial effects of the life-span determinant p66Shc

The 66-kilodalton isoform of the growth factor adapter Shc (p66Shc) translates oxidative damage into cell death by acting as reactive oxygen species (ROS) producer within mitochondria. However, the signaling link between cellular stress and mitochondrial proapoptotic activity of p66Shc was not known. We demonstrate that protein kinase C beta, activated by oxidative conditions in the cell, induces phosphorylation of p66Shc and triggers mitochondrial accumulation of the protein after it is recognized by the prolyl isomerase Pin1. Once imported, p66Shc causes alterations of mitochondrial Ca2+ responses and three-dimensional structure, thus inducing apoptosis. These data identify a signaling route that activates an apoptotic inducer shortening the life span and could be a potential target of pharmacological approaches to inhibit aging.     

运动、胸腺细胞凋亡与机制研究

主要通过文献资料法,对细胞凋亡的机制及运动与胸腺细胞凋亡的关系等进行了述评,其中细胞凋亡与死亡受体信号转导途径、细胞凋亡与线粒体凋亡途径和细胞凋亡与内质网凋亡途径是本文论述的重点。结果显示,有关胸腺细胞凋亡的死亡受体信号转导途径机制和线粒体机制研究不多,有关胸腺细胞凋亡的内质网机制甚少。现有研究显示运动诱导胸腺细胞凋亡与线粒体凋亡途径关系密切,其他有待进一步研究证实。     

Caspases 3 and 7: key mediators of mitochondrial events of apoptosis

The current model of apoptosis holds that upstream signals lead to activation of downstream effector caspases. We generated mice deficient in the two effectors, caspase 3 and caspase 7, which died immediately after birth with defects in cardiac development. Fibroblasts lacking both enzymes were highly resistant to both mitochondrial and death receptor-mediated apoptosis, displayed preservation of mitochondrial membrane potential, and had defective nuclear translocation of apoptosis-inducing factor (AIF). Furthermore, the early apoptotic events of Bax translocation and cytochrome c release were also delayed. We conclude that caspases 3 and 7 are critical mediators of mitochondrial events of apoptosis.