Two phases of intracellular reactive oxygen species production during victorin-induced cell death in oats

Reactive oxygen species (ROS) are thought to be involved in various forms of programmed cell death (PCD) in animal and plant cells. PCD, along with the production of ROS, occurs during plant–pathogen interactions. Here we show that victorin, a host-specific toxin produced by Cochliobolus victoriae, which causes victoria blight of oats, induces two phases of intracellular ROS production in victorin-sensitive oat mesophyll cells. The initial production of ROS is restricted at mitochondria and not accompanied with cellular oxidative damage. Later production of ROS is dispersed into cells concomitant with lipid peroxidation, chloroplast dysfunction, and cell death. Superoxide dismutase can clearly suppress the initial ROS production and delay the progression of cell death. These data indicate that the initial ROS production may be involved in the cell death induction process, and the later ROS production may play important roles in events leading to cellular disruption.     

Methylparathion- and carbofuran-induced mitochondrial dysfunction and oxidative stress in Helicoverpa armigera (Noctuidae: Lepidoptera)

The cotton bollworm, Helicoverpa armigera is a polyphagous pest of several crops in Asia, Africa, and the Mediterranean Europe. Organophosphate and carbamate insecticides are used on a large-scale to control Helicoverpa. Therefore, we studied the effect of methylparathion and carbofuran, an organophosphate and carbamate insecticide, respectively, on oxidative phosphorylation and oxidative stress in H. armigera larvae to gain an understanding of the different target sites of these insecticides. It was observed that state III and state IV respiration, respiratory control index (RCI), and P/O ratios were inhibited in a dose-dependent manner by methylparathion and carbofuran under in vitro and in vivo conditions. Methylparathion and carbofuran inhibited complex II by ∼45% and 30%, respectively. Lipid peroxidation, H₂O₂ content, and lactate dehydrogenase (LDH) activity increased and glutathione reductase (GR) activity decreased in a time- and dose-dependent manner in insecticide-fed larvae. However, catalase activity was not affected in insecticide-fed larvae. Larval growth decreased by ∼64% and 67% in larvae fed on diets with 100 μM of methylparathion and carbofuran. The results suggested that both the insecticides impede the mitochondrial respiratory functions and induced lipid peroxidation, H₂O₂, and LDH leak, leading to oxidative stress in cells, which contribute to deleterious effects of these insecticides on the growth of H. armigera larvae, along with their neurotoxic effects.     

Effect of CREB-shRNA on mitochondrial morphology and cell apoptosis in OGD/R-induced cortical neurons

AIM: To construct recombinant lentiviral vector with short hairpin RNA (shRNA) of CREB gene, and to investigate the effect of CREB gene silencing on mitochondrial morphology and cell apoptosis in oxygen-glucose deprivation/reoxygenation (OGD/R)-induced cortical neurons. METHODS: Three lentiviral vectors pLentiLox3.7 (PLL) inserted shRNA fragments targeting CREB gene were co-transfected with the packaging plasmids psPAX2 and pMD2.G to the 293T cells, and the virus particles, which was infected with the primary cortical neurons, was encapsulated. The protein expression of CREB was detected by Western blot. The mitochondrial morphology, cell apoptosis and the expression of Bcl-2 and Bax were evaluated by the methods of MitoTracker red, TUNEL and Western blot in OGD/R induced cortical neurons after CREB gene silencing. RESULTS: The pLL-CREB-shRNA1 was the most effective shRNA, which inhibited 80% CREB gene expression in the cortical neurons. The mitochondrial was appeared dot and fragment morphology in OGD/R induced cortical neurons with transfected pLL-CREB-shRNA1 plasmid. In addition, the expression of Bcl-2 was decreased, the expression of Bax, and the apoptosis of the neurons were increased by tranfected with pLL-CREB-shRNA1. CONCLUSION: CREB shRNA recombinant lentiviral vector specifically inhibits the expression of CREB gene. CREB gene silencing promotes the cell apoptosis and mitochondrial morphological changes in the cortical neurons induced by OGD/R.     

Effects of uric acid on mitochondrial damage and Pgam5/Drp1 expression in rat kidney cells

AIM To investigate the effects of uric acid on mitochondrial damage and the expression of phosphoglycerate mutase family member 5 (Pgam5) and dynamin-related protein 1 (Drp1) in rat kidney cells. METHODS Normal rat kidney NRK-52E cells were treated with uric acid at 0.6 mmol/L. The cell viability was measured by MTT assay. The cell morphological change was observed by Hoechst 33258 staining. The apoptosis of the cells was analyzed by flow cytometry. The morphological structure of mitochondria was observed under transmission electron microscope. The expression of Pgam5 and Drp1 was examined by immunofluorescence staining. The mRNA expression of Pgam5 in mitochondria was detected by RT-qPCR. The protein expression of Pgam5 and Drp1 in the cytoplasmic matrix was determined by Western blot. RESULTS Uric acid significantly decreased the viability of NRK-52E cells, and significantly increased the apoptotic rate of the cells (P<0.01). Mitochondrial swelling, vacuolation and cristal rupture were observed after the NRK-52E cells were treated with uric acid, and the mRNA expression of Pgam5 in mitochondria was decreased significantly, while the protein expression of Pgam5 and Drp1 in the cytoplasmic matrix was increased significantly (P<0.01). CONCLUSION Uric acid intervention destroys the mitochondrial structure of rat renal cells, up-regulates the expression of Pgam5 and Drp1 in the cytoplasmic matrix, and induces apoptosis of the cells.     

Roles of mitochondrial dysfunction in cardiovascular diseases

Mitochondria are important organelles of energy generation in eukaryocytes and play a pivotal role in cell calcium homeostasis, signal transduction and apoptotic regulation. The possible causes leading to mitochondrial dysfunction include oxidative stress, Ca2+ disorder, reduction of mitochondrial biosynthesis and mitochondrial DNA mutations, all of which are also closely related to the development of cardiovascular diseases. Understanding the mitochondrial dysfunction and its important role in cardiovascular diseases are very significant for elucidating the mechanisms of cardiovascular diseases.     

铝对花生根尖线粒体膜脂 过氧化和有机酸分泌的影响

以耐铝性不同的2个花生品种桂花21 (耐铝)和桂花23 (铝敏感)为材料,研究了铝对花生根尖线粒体膜 脂过氧化、保护酶活性和有机酸分泌的影响。结果表明,桂花23根尖铝含量较桂花21高,苏木精染色深;桂花21 和桂花23根尖线粒体O2 ·、MDA含量在试验范围内随着铝浓度的增加而上升,但桂花21增加的幅度低于桂花23; 根尖线粒体SOD及POD的活性随着铝浓度的增加而增加,桂花21上升的幅度明显大于桂花23。铝诱导花生根系 柠檬酸和琥珀酸分泌显著增加,桂花23柠檬酸增加幅度较琥珀酸高,而桂花21中琥珀酸增加幅度较柠檬酸高。     

杂色角孔海胆形态学观察、亲缘关系及性腺营养成分分析

本研究以杂色角孔海胆(Salmacis sphaeroides variegate)为研究材料,观察其形态,基于线粒体16S rDNA基因部分序列分析杂色角孔海胆与其他棘皮动物的亲缘关系,同时分析杂色角孔海胆性腺中的营养成分,为杂色角孔海胆种质改良和资源评估积累数据。结果显示,杂色角孔海胆的主要形态特征具有口器较小,每个步带板的外缘有3~4个小的圆形凹孔,管足壁内的骨片为C形。通过Mega 5.0对其他棘皮动物和得到的杂色角孔海胆的线粒体16S rDNA基因部分序列进行比较,基于距离法(NJ)和最大似然法(MP)分别构建系统树。结果显示,杂色角孔海胆与疏棘角孔海胆(Salmacis bicolor rarispina)的亲缘关系最近,其次是刻肋海胆属的3种海胆;杂色角孔海胆性腺中水分为86.1%,灰分为3.0%,粗脂肪为2.0%,蛋白质为8.01%;杂色角孔海胆性腺中的氨基酸种类共有16种,总含量为38.71 g/100 g,EAA/TAA为36.99%,EAA/NEAA为71.84%,DAA/TAA为37.17%;脂肪酸共检测到15种,含量最高的是棕榈酸C16:0、硬脂酸C18:0、EPA C20:5和花生四烯酸C20:4,含量最低的为二高-γ-亚麻酸C20:3,较其他海胆,其DHA含量较高。研究结果将为杂色角孔海胆的生物学研究提供基本资料,可用于经济海胆开发应用,同时也可为远缘杂交育种及经济海胆的种质鉴定提供理论依据。     

Advances in mitochondria injury by Aβ in Alzheimer's disease

The β-amyloid protein (Aβ) has long been considered to associate with Alzheimers disease (AD). In addition, groups of evidence show that the soluble intracellular Aβ plays an important role in the disease development. The mitochondrial dysfunction induced by Aβ accumulation is a main pathologic process in early stage of AD. Matured Aβ is imported into the mitochondria through an unclear route. Once inside the mitochondria, Aβ is able to interact with a number of targets, including amyloid-binding alcohol dehydrogenase (ABAD) and cyclophilin D (CypD), which is a component of the mitochondrial permeability transition pore. Interference with the normal functions of these proteins results in mitochondrial injury, such as energy dyshomeostasis, production of reactive oxygen species, membrane permeability alteration and so on. This review explores the Aβ generation and location in mitochondria. The mitochondrial injury induced by the interaction between Aβ and its targets are also discussed.     

硝普钠对低温胁迫下枇杷幼果线粒体AsA-GSH循环代谢的影响

探讨外源一氧化氮供体硝普钠(sodium nitroprusside,SNP)处理对低温胁迫下"早钟6号"枇杷(Eriobotrya japonica Lindl.cv.Zaozhong No.6)幼果线粒体抗氧化系统的影响。以不同浓度的SNP处理"早钟6号"枇杷幼果后再经-3℃低温胁迫处理,测定幼果线粒体的抗氧化物质和抗氧化酶活性的变化。结果表明,低温胁迫下,经0.2和0.5 mmol/L SNP处理的枇杷幼果线粒体过氧化氢(H2O2)和丙二醛(MDA)含量均低于对照(CK),而还原型谷胱甘肽(GSH)和抗坏血酸(AsA)含量以及抗坏血酸过氧化物酶(APX,EC 1.11.1.11)、谷胱甘肽还原酶(GR,EC 1.6.4.2)、脱氢抗坏血酸还原酶(DHAR,EC 1.8.5.1)和单脱氢抗坏血酸还原酶(MDHAR,EC 1.6.5.4)活性均高于CK,但采用1.0 mmol/L较高浓度的SNP处理对提高枇杷幼果抗氧化能力的作用不明显。适当的外源NO处理可提高枇杷幼果线粒体GSH和AsA抗氧化剂的含量以及APX、GR、DHAR和MDHAR酶的活性,降低H2O2和MDA含量,减轻细胞的氧化损伤,增强枇杷幼果在低温胁迫下的抗寒能力。