Involvement of mitochondrial-mediated caspase-3 activation and lysosomal labilization in acrylamide-induced liver toxicity

Acrylamide (ACR) is a chemical frequently used in both industrial and synthetic processes and may be produced during food processing. ACR at very high concentrations is postulated to exert its toxicity through the stimulation of an oxidative stress. ACR in excessive doses induces the central nervous system, reproduction, and genetic toxicity. However, ACR effects on the liver, a major organ of drug metabolism, have not been adequately explored. In addition, the role of mitochondria in an ACR-mediated hepatotoxicity is still unclear. The aim of this study was to investigate the cytotoxic mechanisms attributed to ACR using isolated rat hepatocytes. Hepatocytes were isolated by the collagenase perfusion method and incubated with an EC50_2hr concentration of ACR for 3 hr. The EC50_{2 hr} of ACR on isolated rat hepatocytes was determined to be 1 mM. Based on our results, hepatocytes cytotoxicity of ACR (1 mM) was mediated by a reactive oxygen species formation and lipid peroxidation. Incubation of hepatocytes with ACR produced rapid hepatocyte glutathione depletion which is another marker of the cellular oxidative stress. ACR cytotoxicity was also associated with mitochondrial injury as evidenced by the decline of mitochondrial membrane potential and lysosomal membrane leakiness. Our results also showed that ACR induced caspase-3 activation, the final mediator of apoptosis signaling. These findings contribute to a better understanding underlying mechanisms involved in ACR hepatotoxicity originating from the oxidative stress and ending in mitochondrial/lysosomal damage and cell death signaling.     

植物线粒体基因转录水平表达研究方法的建立

以茎瘤芥雄性不育系为材料,在已有文献的基础上,对常规方法进行改进,使植物线粒体分离、线粒体切片观察、线粒体RNA提取、反转录方法、PCR扩增、northern杂交等方法系统化,建立了一整套比较有效的适合于植物线粒体基因转录水平表达分析的方法.图4参12     

Direct toxicity of amyloid beta peptide on rat brain mitochondria: preventive role of Mangifera indica and Juglans regia

Role of mitochondrial dysfunction and oxidative stress has been well documented in various cognitive-related disorders such as Alzheimer's disease (AD). Evidence indicates that Aß formation impairs mitochondrial function and that mitochondrial dysfunction is an early event in the pathogenesis of AD. The present study was, therefore, designed to investigate the direct toxicity of Aß peptide on isolated mitochondria obtained from rat brain. Various mitochondrial toxicity/integrity parameters such as succinate dehydrogenase activity, reactive oxygen species (ROS) formation, mitochondrial membrane potential collapse (MMP), mitochondrial swelling, and cytochrome c release were measured following the addition of Aß peptide on isolated mitochondria and then, mitoprotective effect of aqueous extracts of Mangifera indica and Juglans regia against mitochondrial toxicity endpoints parameters induced by Aß peptide were assessed. Our results showed that exposure to Aß peptide (30 nM) in isolated brain mitochondria induced mitochondrial ROS formation, MMP collapse, mitochondrial swelling, and cytochrome c release which is the starting point of apoptosis signaling. All these mitochondrial toxic endpoints induced by Aß peptide inhibited by aqueous extracts of Mangifera indica (100–400 µg/ml) and Juglans regia (200–400 µg/ml). To our knowledge, this is one of the first apparent studies to claim directly targeting of brain mitochondria and induction of apoptosis by Aß peptide as a new hypothesis for etiology of AD and other related neurodegenerative diseases as well as mitopreventive role of common antioxidant nutritional products including walnut and mango.     

Antimony induces oxidative stress and cell death in normal hepatocytes

Antimony (Sb) accumulates in the liver which is one of the target organs for metal-mediated toxicity. Although toxicity of Sb was previously investigated, the precise mechanism of Sb-induced hepatotoxicity remains to be determined. The aim of this study was to examine the role of oxidative stress, and mitochondria in the induction of cell death by Sb. Our results showed that liver cell lysis induced by Sb is mediated by reactive oxygen species (ROS) formation, lipid peroxidation and decline of mitochondrial membrane potential (MMP). Antimony-induced ROS formation, lipid peroxidation and reduction of MMP were significantly diminished by antioxidants and ROS scavengers such as dimethyl sulfoxide and mannitol; mitochondrial permeability transition (MPT) pore sealing agents such as carnitine and trifluoperazine; and adenosine triphosphate (ATP) generator, L-glutamine. Antimony-induced ROS formation, lipid peroxidation and fall in MMP were potentiated by glutathione (GSH) depletion via n-bromoheptane. MPT pore sealing agents and ATP generator inhibited hepatotoxicity, indicating Sb-activated cell death via mitochondrial pathway. Pretreatment of hepatocytes with antioxidants and ROS scavengers also blocked cell death induced by Sb, whereas GSH depletion enhances Sb-induced cell death, suggesting that oxidative stress may be directly involved in the reduction of MMP. These findings contribute to a better understanding of the mechanisms that mediate Sb-induced cell death in isolated rat hepatocytes.     

镧、铈、钕对小鼠肝细胞线粒体的氧化损伤作用

轻稀土元素进入生物体后主要累积于肝脏,进入肝细胞,除蓄积在细胞核中,还存在于线粒体中。为探讨轻稀土元素对小鼠肝细胞线粒体的氧化损伤作用,选用5周龄雄性ICR小鼠分别以10、20和40mg·kg~(-1)的镧(La)、铈(Ce)和钕(Nd)灌胃,6周后测定小鼠肝细胞线粒体中超氧化物岐化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GPx)的活性,以及谷胱甘肽(GSH)和丙二醛(MDA)的含量。结果显示,与对照组相比,La中剂量组和Ce低剂量组SOD活性显著升高,La高剂量组和Nd中、高剂量组中SOD活性显著降低(P<0.05,P<0.01);除个别剂量组外,各染毒组CAT和GPx活性与GSH含量显著降低(P<0.05,P<0.01);Nd各剂量组、La高剂量组和Ce高剂量组的MDA含量显著升高(P<0.05,P<0.01)。研究表明,La、Ce和Nd所导致的CAT和GPx活性以及GSH含量降低可能是造成肝细胞线粒体氧化损伤的主要原因。     

细胞凋亡的线粒体途径调控

细胞凋亡是近年来研究的一个热点问题，涉及到细胞内许多复杂的生化过程．线粒体是真核生物能量和代谢的中心，也是细胞凋亡信号传导途径中起关键调节作用的细胞器，对细胞凋亡的线粒体途径调控机制进行研究具有重要意义．本文综述了细胞凋亡过程中线粒体途径的信号传导及其调控机制、线粒体对细胞凋亡信号的反应、细胞凋亡过程中线粒体功能丧失几方面的研究进展．图1参43     

The Structure of Energy Supply for Tissue Adaptation in Small Mammals Differing in Ecological Specialization

Multivariate factor analysis of the data on liver bioenergetics of mountain and forest voles has been performed. The specific features of tissue energy metabolism are best differentiated in the space of characters pertaining to the succinic acid oxidation system. The role of the dominant oxidation substrate in the structure of energy supply for the adaptation of tissue to a set of environmental factors in specialized and widespread species is discussed.     

六氯苯对离体鱼肝线粒体抗氧化酶的作用

采用差速离心法从鲫鱼肝脏中提取线粒体,用不同浓度(0,2,4,8,16,32mg/L)六氯苯对其体外染毒30min.测定线粒体超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)活性,用聚丙烯酰胺凝胶电泳法分析其同工酶谱,并检测线粒体中丙二醛(MDA)含量.结果显示,SOD和GSH-Px活性及其同工酶的活性表达均呈现出低浓度六氯苯作用下被激活,高浓度六氯苯作用下被抑制的变化趋势.在高浓度六氯苯(32mg/L)作用下线粒体中MDA含量显著增加.说明六氯苯的毒性作用可能为一种自由基机制,即低浓度的六氯苯导致线粒体内活性氧自由基(ROS)生成量少量增加,SOD和GSH-Px及其同工酶活性由于氧化应激的诱导被激活;随着六氯苯浓度增加,线粒体内ROS生成量大量增加,并破坏了SOD和GSH-Px的抗氧化活性,导致其活力下降或丧失,自由基含量增加,线粒体脂质过氧化加剧.