RESUMO
Radiation-induced damage to the mitochondrial macromolecules and electron transfer chain (ETC), causing the generation of primary and secondary reactive oxygen (ROS) species. The continuous ROS production after radiation will trigger cell oxidative stress and ROS-mediated nucleus apoptosis and autophagy signaling pathways. Scavenging radiation-induced ROS effectively can help mitochondria to maintain their physiological function and relief cells from oxidative stress. Nicotinamide is a critical endogenous antioxidant helping to neutralize ROS in vivo. In this study, we designed and synthetized a novel mitochondrial-targeted dihydronicotinamide (Mito-N) with the help of mitochondrial membrane potential to enter the mitochondria and scavenge ROS. According to experiment results, Mito-N significantly increased cell viability by 30.75% by neutralizing the accumulated ROS and resisting DNA strands breaks after irradiation. Furthermore, the mice survival rate also improved with the treatment of Mito-N, by effectively ameliorating the hematopoietic system infliction under lethal dose irradiation.
Assuntos
Mitocôndrias/efeitos dos fármacos , Niacinamida/análogos & derivados , Protetores contra Radiação/farmacologia , Animais , Antioxidantes/farmacologia , Células CHO , Cricetinae , Cricetulus , Desenho de Fármacos , Masculino , Camundongos Endogâmicos C57BL , Niacinamida/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Lesões Experimentais por Radiação/prevenção & controle , Espécies Reativas de Oxigênio/metabolismoRESUMO
Human K(v) channel interacting protein 1 (KCHIP1) is a new member of the neural calcium binding protein superfamily. Theoretically KCHIP1 has several calcium binding domains and two myristoylation sites. In this study, we demonstrated that the calcium binding domains and myristoylation sites were functional. The first, through running SDS-PAGE gel, we testified its ability of binding Ca(2+) with obvious discrepancy of the electrophoresis migrating rate after binding Ca(2+). Then, through the techniques of fused green fluorescence protein and site-directed mutagenesis, we demonstrated that wild type KCHIP1 protein accumulated in the secretory vesicles of Golgi body. In contrast, its two mutated forms without myristoylation sites accumulated throughout the whole cytoplasm. These observations indicate that KCHIP1 protein has a myristoylation motif mediating the interaction between KCHIP1 protein and membrane.
Assuntos
Proteínas de Ligação ao Cálcio/metabolismo , Proteínas Interatuantes com Canais de Kv/química , Proteínas Interatuantes com Canais de Kv/fisiologia , Canais de Potássio/metabolismo , Animais , Células COS , Cálcio/metabolismo , Proteínas de Ligação ao Cálcio/genética , Chlorocebus aethiops , Humanos , Canais de Potássio de Abertura Dependente da Tensão da Membrana/metabolismo , Transporte Proteico , Proteínas Recombinantes de Fusão/metabolismo , TransfecçãoRESUMO
O(-.)(2) production based on chemical method by Ecc-treated intact tobacco leaves was observed. It showed a long-lasting one-phase course beginning 1 h and ending 14 h after Erwinia carotovora subsp. carotovora (Ecc) was inoculated. In Ecc-treated leaves, O(-.)(2) production rate of 3 h is 2 times of 1 h, and of 10 h it remains 1.7 times. It decreases rapidly between 10 h and 14 h, at 14 h it falls to the level before inoculation. This is a completely different course from the widely accepted two-phase course obtained from suspension-cultured cells. Electron spin resonance (ESR) spectrum of O(-.)(2) produced by intact tobacco leaves after Ecc inoculation 2 h and 6 h was also observed, the amplitude of 6 h is larger than of 2 h in Ecc-treated group. In the control group, there is no difference between the amplitude of 2 h and 6 h. This indicated that O(-.)(2) amount of Ecc-treated group is higher than of the control group, which is the same result as obtained through chemical method. The ESR spectrum of O(-.)(2) produced by chloroplasts and mitochondria from tobacco leaves after 2 h and 6 h after Ecc inoculation were observed. In chloroplast experiment, the amplitude of 6 h was larger than of 2 h in Ecc-treated group while it was the same in the control group. ESR spectrum of O(-.)(2) produced by mitochondria was proved to be a same result after a careful comparison, in spite of a greater difference between 2 h and 6 h in control group than chloroplast experiment. The fact that spectra of these two organelles were both synchronous with that of intact leaves implied that these two organelles possibly participated in cellular O(-.)(2) burst. Chloroplasts in the dark showed no ESR spectrum 2 h and 6 h after Ecc inoculation in Ecc-treated group as well as in the control group, indicating that O(-.)(2) in the chloroplast probably originated from the photosynthetic electron transport process.
Assuntos
Nicotiana/microbiologia , Pectobacterium carotovorum/fisiologia , Folhas de Planta/microbiologia , Espécies Reativas de Oxigênio/metabolismo , Cloroplastos/metabolismo , Espectroscopia de Ressonância de Spin Eletrônica , Interações Hospedeiro-Patógeno , Mitocôndrias/metabolismo , Folhas de Planta/metabolismo , Nicotiana/metabolismoRESUMO
Many phytopathogenic bacteria were founded to release the superoxide anion (O2-) by themselves from different sites and the production of O2- was related to the pathogenicity of strains. The presences of O2- in cytomembrane, cell wall and filtrate were confirmed by chemical method and electron spin resonance (ESR). The results show that filtrate may be the primary site of O2- production due to its higher O2- generating activity either in the presence or the absence of DDC. Additional lines of evidence further suggested a regulatory mechanism of O2- production in phytopathogenic bacteria.
Assuntos
Bactérias/metabolismo , Doenças das Plantas/microbiologia , Superóxidos/metabolismo , Bactérias/patogenicidade , Espectroscopia de Ressonância de Spin Eletrônica , Medições LuminescentesRESUMO
OBJECTIVE: To detect the changes in the expression of apoptosis signals: Fas, FasL and NF-kappa B in the process of osteoclast-like cell (OLC) apoptosis effected by sodium fluoride. METHODS: After co-culture of osteoclast-like cells with 0, 5, 10 and 15 mg/L sodium fluoride, Fas, FasL and NF-kappa B antibody expressions were detected by immune-histochemistry. RESULTS: The expression of Fas and FasL increased with the concentration of the sodium fluoride, however the expression of NF-kappa B decreased with the concentration of sodium fluoride. CONCLUSION: In the process of OLC apoptosis induced by sodium fluoride, the expression of Fas and FasL increased, and that of NF-kappa B decreased with the concentration of sodium fluoride respectively, and the changes of the expression present a dose-dependent pattern.
Assuntos
Apoptose , Fluoretos/farmacologia , Glicoproteínas de Membrana/metabolismo , NF-kappa B/metabolismo , Osteoclastos/citologia , Receptor fas/metabolismo , Animais , Proteína Ligante Fas , Feminino , Ligantes , Camundongos , Camundongos Endogâmicos C57BL , Osteoclastos/metabolismoRESUMO
Taking the Arachis hypogaea cv. 'Qinghua 7' as test material, a field experiment was conducted to study the effects of different cultivation modes on the leaf photosynthetic characteristics and yield of summer-sowing peanut after wheat harvest. As compared with conventional cultivation mode, high-yield protective cultivation mode promoted the leaf growth, significantly improved the leaf area index (LAI), and maintained a longer time of high LAI and chlorophyll content. Meanwhile, the net photosynthetic rate, stomatal conductance, and transpiration rate of functional leaves under high-yield protective cultivation mode were higher while the intercellular CO2 concentration was lower, which induced the photosynthetic efficiency of functional leaves being significantly improved. Therefore, under high-yield protective cultivation mode, the yield per peanut plant was higher, the pod yield increased significantly, and the economic coefficient improved obviously. Both film mulching and straw returning could also improve the leaf photosynthesis of summer-sowing peanut, and increase the peanut yield. It was suggested that high-yield protective cultivation mode could effectively alleviate the adverse factors of summer-sowing peanut, such as the short growth period and lower productivity per plant, being a practical high-yield cultivation mode of summer-sowing peanut.
Assuntos
Agricultura/métodos , Arachis/crescimento & desenvolvimento , Fotossíntese/fisiologia , Folhas de Planta/fisiologia , Arachis/fisiologia , Biomassa , China , Folhas de Planta/crescimento & desenvolvimento , Estações do AnoRESUMO
The present study aimed to find the mutations of KCHIP1 gene in breast cancer. KCHIP1 cDNA samples from 12 specimens of breast cancer and 12 specimens of normal mammary tissues were amplified by reverse transcription-polymerase chain reaction (RT-PCR), and directly sequenced to detect mutation. No mutation of KCHIP1 gene was found in these samples; while a new splicing type of KCHIP1 gene was found, which has an insert (162 bp) between exon 1 and exon 2 in KCHIP1 gene (AY780424).