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Mitochondrial superoxide production during oxalate-mediated oxidative stress in renal epithelial cells 总被引:8,自引:0,他引:8
Khand FD Gordge MP Robertson WG Noronha-Dutra AA Hothersall JS 《Free radical biology & medicine》2002,32(12):2394-1350
Crystals of calcium oxalate monohydrate (COM) in the renal tubule form the basis of most kidney stones. Tubular dysfunction resulting from COM-cell interactions occurs by mechanism(s) that are incompletely understood. We examined the production of reactive oxygen intermediates (ROI) by proximal (LLC-PK1) and distal (MDCK) tubular epithelial cells after treatment with COM (25–250 μg/ml) to determine whether ROI, specifically superoxide (O2•−), production was activated, and whether it was sufficient to induce oxidative stress. Employing inhibitors of cytosolic and mitochondrial systems, the source of ROI production was investigated. In addition, intracellular glutathione (total and oxidized), energy status (ATP), and NADH were measured. COM treatment for 1–24 h increased O2•− production 3–6-fold as measured by both lucigenin chemiluminescence in permeabilized cells and dihydrorhodamine fluorescence in intact cells. Using selective inhibitors we found no evidence of cytosolic production. The use of mitochondrial probes, substrates, and inhibitors indicated that increased O2•− production originated from mitochondria. Treatment with COM decreased glutathione (total and redox state), indicating a sustained oxidative insult. An increase in NADH in COM-treated cells suggested this cofactor could be responsible for elevating O2•− generation. In conclusion, COM increased mitochondrial O2•− production by epithelial cells, with a subsequent depletion of antioxidant status. These changes may contribute to the reported cellular transformations during the development of renal calculi. 相似文献
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Properties of microtubule sliding disintegration in isolated tetrahymena cilia 总被引:5,自引:5,他引:0
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Properties of the sliding disintegration response of demembranated tetrahymena cilia have been studied by measuring the spectrophotomeric response or turbidity of cilia suspensions at a wavelength of 350 nm relative to changes in the dynein substrate (MgATP(2-)) concentration. The maximum decrease in turbidity occurs in 20 muM ATP, and 90 percent of the decrease occurs in approximately 5.9 s. At lower ATP concentrations (1-20 muM), both the velocity and magnitude of the turbidity decreases are proportional to ATP concentration. The velocity data for 20 muM ATP permit construction of a reaction velocity curve suggesting that changes in turbidity are directly proportional to the extent and velocity of disintegration. At ATP concentrations more than 20 muM (50muM to 5mM), both velocity and magnitude of the turbidimetric response are reduced by approximately 50 percent. This apparent inhibition results in a biphasic response curve that may be related to activation of residual shear resistance or regulatory components at the higher ATP concentrations. The inhibitory effects of elevated ATP can be eliminated by mild trypsin proteolysis, whereupon the reaction goes to completion at any ATP concentration. The turbidimetric responses of the axoneme-substrate suspensions are consistent with the extent and type of axoneme disintegration revealed by electron microscope examination of the various suspensions, suggesting that the turbidimetric assay may prove to be a reliable means for assessing the state of axoneme integrity. 相似文献
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Jonathan WF Mant Suzanne H Richards FD Richard Hobbs David Fitzmaurice Gregory YH Lip Ellen Murray Miriam Banting Kate Fletcher Joy Rahman Teresa Allan James Raftery Stirling Bryan 《BMC cardiovascular disorders》2003,3(1):1-10
Background
Statins effectively lower blood cholesterol and the risk of cardiovascular death. Immunomodulatory actions, independent of their lipid-lowering effect, have also been ascribed to these compounds. Since macrophages participate in several vascular pathologies, we examined the effect of statin treatment on the survival and differentiation of primary human monocytes.Methods
Peripheral blood mononuclear cells (PBMCs) from healthy individuals were cultured in the presence or absence of mevastatin. Apoptosis was monitored by annexin V / PI staining and flow cytometry. In parallel experiments, cultures were stimulated with LPS in the presence or absence of mevastatin and the release of IL-1β and IL-1Ra was measured by ELISA.Results
Among PBMCs, mevastatin-treated monocytes were particularly susceptible to apoptosis, which occurred at doses >1 microM and was already maximal at 5 microM. However, even at the highest mevastatin dose used (10 microM), apoptosis occurred only after 24 h of culture, possibly reflecting a requirement for cell commitment to differentiation. After 72 h of treatment the vast majority (>50%) of monocytes were undergoing apoptosis. Stimulation with LPS revealed that mevastatin-treated monocytes retained the high IL-1β output characteristic of undifferentiated cells; conversely, IL-1Ra release was inhibited. Concurrent treatment with mevalonolactone prevented the induction of apoptosis and suppressed both IL-1β and IL-1Ra release in response to LPS, suggesting a rate-limiting role for HMG-CoA reductase in monocyte differentiation.Conclusions
Our findings indicate that statins arrest the functional differentiation of monocytes into macrophages and steer these cells into apoptosis, suggesting a novel mechanism for the vasculoprotective properties of HMG-CoA reductase inhibitors. 相似文献6.
All successive Cretaceous reference horizons (except the uppermost one) are described in one section, the most complete in Mongolia. 相似文献
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Helen A Arcuri Geraldo FD Zafalon Evandro A Marucci Carlos E Bonalumi Nelson JF da Silveira José M Machado Walter F de AzevedoJr Mário S Palma 《BMC bioinformatics》2010,11(1):12
Background
The functional and structural characterisation of enzymes that belong to microbial metabolic pathways is very important for structure-based drug design. The main interest in studying shikimate pathway enzymes involves the fact that they are essential for bacteria but do not occur in humans, making them selective targets for design of drugs that do not directly impact humans. 相似文献8.
The Cretaceous and Tertiary development of Mongolian non-marine ostracod faunas is reviewed. During the Late Cretaceous and Early Palaeogene, representatives of the Cypridoidea were widespread and common, Cytheroidea less so and the Darwinuloidea comparatively rare. The evolutionary history of the subfamily Talicyprideinae is considered, with reference to the genera Talicypridea, Altanicypris, Khandiaand Bogdocypris. It is suggested that the extinct Talicyprideinae were related to the mid-Cretaceous to Recent subfamily Cypridinae (e.g. the genus Cypris), both belonging to the family Cyprididae. It is shown that early representatives of the Cyprididae, one of the most diverse non-marine cypridoidean families today, were present from Early Cretaceous onwards (e.g. Lycopterocypris, Mongolocypris), alongside the dominant Cretaceous cypridoideans, the Cyprideidae (e.g. Cypridea), which became extinct in the Palaeogene. 相似文献
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