Relationships between the mitochondrial transmembrane potential, ATP concentration, and cytotoxicity in isolated rat hepatocytes

Wu, Ellen; Smith, Martyn T.; Bellomo, Giorgio; Monte, Donato Di

doi:10.1016/0003-9861(90)90129-m

Cited by 133 publications

(46 citation statements)

References 19 publications

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“…Disruption of electron transport, oxidative phosphorylation, and ATP production are known as one of the general mechanisms in triggering apoptotic and nonapoptotic cell death by mitochondria (Green and Reed, 1998). Wu et al (1990) considered that a loss of mitochondrial membrane potential (MMP) might occur only when supplies of ATP were depleted. So we confirmed that the impairment of ETC might be one of the very earlier events of apoptosis induced by MCs.…”

Section: Discussionmentioning

confidence: 99%

In vivo studies on the toxic effects of microcystins on mitochondrial electron transport chain and ion regulation in liver and heart of rabbit

Zhao

Xie

Tang

et al. 2008

Comparative Biochemistry and Physiology Part C: Toxicology &

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Section: Discussionmentioning

confidence: 99%

In vivo studies on the toxic effects of microcystins on mitochondrial electron transport chain and ion regulation in liver and heart of rabbit

Zhao

Xie

Tang

et al. 2008

Comparative Biochemistry and Physiology Part C: Toxicology &

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“…2, 8, 12, 16, or 32 M Rh123 was added to the incubation buffer during the last 10 min of fura-2 loading, and the coverslips were then washed. In studies of hepatocytes, the concentration of Rh123 reaches equilibrium across the plasma membrane after incubation of the cells for 10 min in a buffer containing the probe (16). Measurements were carried out with a SPEX fluorolog-2 CM1T11I system connected to an inverted microscope (Zeiss, Axiovert 35 M).…”

Section: Methodsmentioning

confidence: 99%

Glucose-induced Oscillations in Cytoplasmic Free Ca2+Concentration Precede Oscillations in Mitochondrial Membrane Potential in the Pancreatic β-Cell

Kindmark

Köhler

Brown

et al. 2001

Journal of Biological Chemistry

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Using dual excitation and fixed emission fluorescence microscopy, we were able to measure changes in cytoplasmic free Ca 2؉ concentration ([Ca 2؉ ] i ) and mitochondrial membrane potential simultaneously in the pancreatic ␤-cell. The ␤-cells were exposed to a combination of the Ca 2؉ indicator fura-2/AM and the indicator of mitochondrial membrane potential, rhodamine 123 (Rh123 [Ca 2ϩ ] i in pancreatic ␤-cells using the dye fura-2 (4, 6, 7). Measurements of mitochondrial membrane potential in these cells, using the lipophilic cationic dye rhodamine 123 (Rh123), have also been reported (8). Simultaneous measurements of [Ca 2ϩ ] i and mitochondrial membrane potential in the pancreatic ␤-cell would be of great interest and could provide important information about the possible correlation between mitochondrial responses to glucose stimulation and events at the plasma membrane of the ␤-cell. Such simultaneous measurements have recently been performed in neurons (9, 10) and in mouse ␤-cells (11). To investigate the relationship between mitochondrial membrane potential and [Ca 2ϩ ] i in the ␤-cell, we loaded ␤-cells with fura-2 and Rh123 and applied the method of simultaneous measurements of these two parameters. Our methodological evaluation included an investigation of cellular Rh123 distribution as observed with confocal microscopy. We also estimated the contribution to emitted light from both dyes at the respective excitation wavelength, allowing us to correct for contaminating fluorescence. EXPERIMENTAL PROCEDURESAll reagents were of analytical grade and Millipore water was used. Bovine serum albumin fraction V was from Sigma, collagenase was from Roche Molecular Biochemicals, and fura-2/AM as well as Rh123 were from Molecular Probes Europe BV (Leiden, The Netherlands). Statistical summaries of data are expressed as the mean Ϯ S.E. Where appropriate, the possible statistical significance of differences between two sets of data was tested using Student's t test for unpaired data. In all experimental protocols, ␤-cells from at least three different animals were used, if not otherwise specified.Animals and Preparation of Cells-Adult obese mice (gene symbol ob/ob) of both sexes were obtained from a local colony and starved for 24 h. The animals were killed by decapitation and the islets isolated using a collagenase technique (12). The islets of these mice contain more than 90% ␤-cells (13). A cell suspension was prepared and washed essentially as described previously (14). The cells were suspended in RPMI 1640 culture medium (Flow Laboratories) containing 11 mM glucose supplemented with 10% (v/v) fetal bovine serum, 2 mM Lglutamine, 50 IU/ml penicillin, and 50 g/ml streptomycin (Life Technologies, Inc.). The cell suspension was seeded onto coverslips, and the cells allowed to attach for 2 h and then cultured for up to 3 days in the above medium.Media-The basal medium used for preparation of cells as well as experiments was a HEPES buffer, pH 7.4, with Cl Ϫ as the sole anion (15), containing 3 mM glucose, 1.28 ...

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“…As for the evidence that nimesulide stimulates state 4 respiration and decreases membrane potential of mitochondria it seems likely that ATP depletion results mainly from the uncoupling of oxidative phosphorylation in the hepatocytes. Accordingly, when fructose, a substrate for the glycolytic pathway in liver that prevents ATP depletion and cell damage induced by toxic compounds (Wu et al, 1990;Nieminen et al, 1994), was added to the cell suspension, a significant protection against both the nimesulide-induced injury of hepatocytes and ATP level decrease occurred, and the additional presence of oligomycin stressed this effect. On the other hand, the lack of a significant protection of cyclosporin A against cell injury indicates that MPT onset (Zoratti and Szabò, 1995), as observed in isolated mitochondria, is not significantly involved.…”

Section: Toxicity Of Nimesulide To Mitochondria Of Hepatocytes 605mentioning

confidence: 99%

The Critical Role of Mitochondrial Energetic Impairment in the Toxicity of Nimesulide to Hepatocytes

Mingatto

Rodrigues²,

Pigoso³

et al. 2002

J Pharmacol Exp Ther

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We described the effects of nimesulide (N-[4-nitro-2-phenoxyphenyl]-methanesulfonamide) and its reduced metabolite in isolated rat hepatocytes. Nimesulide stimulated the succinatesupported state 4 respiration of mitochondria, indicating an uncoupling effect of the drug. Incubation of hepatocytes with nimesulide (0.1-1 mM) elicited a concentration-and time-dependent decrease in cell viability as assessed by lactate dehydrogenase leakage, a decrease of mitochondrial membrane potential as assessed by rhodamine 123 retention, and cell ATP depression. Nimesulide also decreased the levels of NAD(P)H and glutathione in hepatocytes, but the extent of the effects was less pronounced in relation to the energetic parameters; in addition, these effects did not imply the peroxidation of membrane lipids. The decrease in the viability of hepatocytes was prevented by fructose and, to a larger extent, by fructose plus oligomycin; it was stimulated by proadifen, a cytochrome P450 inhibitor. In contrast, the reduced metabolite of nimesulide did not present any of the effects observed for the parent drug. These results indicate that: 1) nimesulide causes injury to the isolated rat liver cells, 2) this effect is mainly mediated by impairment of ATP production by mitochondria due to uncoupling, and 3) on account of the activity of its nitro group, the parent drug by itself is the main factor responsible for its toxicity to the hepatocytes.

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Relationships between the mitochondrial transmembrane potential, ATP concentration, and cytotoxicity in isolated rat hepatocytes

Cited by 133 publications

References 19 publications

In vivo studies on the toxic effects of microcystins on mitochondrial electron transport chain and ion regulation in liver and heart of rabbit

In vivo studies on the toxic effects of microcystins on mitochondrial electron transport chain and ion regulation in liver and heart of rabbit

Glucose-induced Oscillations in Cytoplasmic Free Ca2+Concentration Precede Oscillations in Mitochondrial Membrane Potential in the Pancreatic β-Cell

The Critical Role of Mitochondrial Energetic Impairment in the Toxicity of Nimesulide to Hepatocytes

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