PubMed Health⌕ Search

Biomedical subjects

P Ghafourifar

Publications and source records attributed to P Ghafourifar.

At least 19 recordsLinked to original sources
PubMed2001

S-Nitrosylation of mitochondrial caspases.

Caspase-3 is a cysteine protease located in both the cytoplasm and mitochondrial intermembrane space that is a central effector of many apoptotic pathways. In resting cells, a subset of caspase-3 zymogens is S-nitrosylated at the active site cysteine, inhibiting enzyme activity. During Fas-induced apoptosis, caspases are denitrosylated, allowing the catalytic site to function. In the current studies, we sought to identify the subpopulation of caspases that is regulated by S-nitrosylation. We report that the majority of mitochondrial, but not cytoplasmic, caspase-3 zymogens contain this inhibitory modification. In addition, the majority of mitochondrial caspase-9 is S-nitrosylated. These studies suggest that S-nitrosylation plays an important role in regulating mitochondrial caspase function and that the S-nitrosylation state of a given protein depends on its subcellular localization.

Caspase 3↗
PubMed2001

Redox control of mitochondrial functions.

Redox reactions and electron flow through the respiratory chain are the hallmarks of mitochondria. By supporting oxidative phosphorylation and metabolite transport, mitochondrial redox reactions are of central importance for cellular energy conversion. In the present review, we will discuss two other aspects of the mitochondrial redox state: (i) its control of mitochondrial Ca2+ homeostasis, and (ii) the intramitochondrial formation of reactive oxygen or nitrogen species that strongly influence electron flow of the respiratory chain.

Animals↗
PubMed2000

Peroxynitrite formed by mitochondrial NO synthase promotes mitochondrial Ca2+ release.

Mitochondria contribute to the maintenance of the intracellular Ca2+ homeostasis by taking up and releasing the cation via separate and specific pathways. The molecular details of the release pathway are elusive but its stimulation by the cross-linking of some vicinal thiols and consequently NAD+ hydrolysis are known. Thiol cross-linking and NAD+ hydrolysis can be achieved by addition of peroxynitrite (ONOO-), the product of the reaction between superoxide (O2-) and nitric oxide (nitrogen monoxide, NO*) to mitochondria. Mitochondria contain an NO synthase (mtNOS), which is stimulated by Ca2+, and are a copious source of O2-. We show here that intramitochondrially formed ONOO- stimulates the specific, NAD+-linked Ca2+ release from mitochondria. Our findings that upon Ca2+ uptake mtNOS is stimulated, that ONOO- is formed, and that Ca2+ is subsequently released from intact mitochondria suggest the existence of a feedback loop, which prevents overloading of mitochondria with Ca2+.

Animals↗
PubMed1999

Mitochondrial nitric-oxide synthase stimulation causes cytochrome c release from isolated mitochondria. Evidence for intramitochondrial peroxynitrite formation.

Nitric oxide (NO) is synthesized by members of the NO synthase (NOS) family. Recently the existence of a mitochondrial NOS (mtNOS), its Ca(2+) dependence, and its relevance for mitochondrial bioenergetics was reported (Ghafourifar, P., and Richter, C. (1997) FEBS Lett. 418, 291-296; Giulivi, C., Poderoso, J. J., and Boveris, A. (1998) J. Biol. Chem. 273, 11038-11043). Here we report on the possible involvement of mtNOS in apoptosis. We show that uptake of Ca(2+) by mitochondria triggers mtNOS activity and causes the release of cytochrome c from isolated mitochondria in a Bcl-2-sensitive manner. mtNOS-induced cytochrome c release was paralleled by increased lipid peroxidation. The release of cytochrome c as well as increase in lipid peroxidation were prevented by NOS inhibitors, a superoxide dismutase mimic, and a peroxynitrite scavenger. We show that mtNOS-induced cytochrome c release is not mediated via the mitochondrial permeability transition pore because the release was aggravated by cyclosporin A and abolished by blockade of mitochondrial calcium uptake by ruthenium red. We conclude that, upon Ca(2+)-induced mtNOS activation, peroxynitrite is formed within mitochondria, which causes the release of cytochrome c from isolated mitochondria, and we propose a mechanism by which elevated Ca(2+) levels induce apoptosis.

Animals↗
PubMed1999

Ceramide induces cytochrome c release from isolated mitochondria. Importance of mitochondrial redox state.

In the present study we show that N-acetylsphingosine (C2-ceramide), N-hexanoylsphingosine (C6-ceramide), and, to a much lesser extent, C2-dihydroceramide induce cytochrome c (cyto c) release from isolated rat liver mitochondria. Ceramide-induced cyto c release is prevented by preincubation of mitochondria with a low concentration (40 nM) of Bcl-2. The release takes place when cyto c is oxidized but not when it is reduced. Upon cyto c loss, mitochondrial oxygen consumption, mitochondrial transmembrane potential (Delta Psi), and Ca2+ retention are diminished. Incubation with Bcl-2 prevents, and addition of cyto c reverses the alteration of these mitochondrial functions. In ATP-energized mitochondria, ceramides do not alter Delta Psi, neither when cyto c is oxidized nor when it is reduced, ruling out a nonspecific disturbance by ceramides of mitochondrial membrane integrity. Furthermore, ceramides decrease the reducibility of cyto c. We conclude that the apoptogenic properties of ceramides are in part mediated via their interaction with mitochondrial cyto c followed by its release and that the redox state of cyto c influences its detachment by ceramide from the inner mitochondrial membrane.

Animals↗
PubMed1999

Increase by NO synthase inhibitor of lead-induced release of N-acetyl-beta-D-glucosaminidase from perfused rat kidney.

Urinary N-acetyl-beta-D-glucosaminidase (NAG) had been shown to be a useful early marker of renal injury such as lead nephrotoxicity. This study investigated the effect of lead acetate on nephrotoxicity and its correlation with the nitric oxide (NO) system by determining the NAG release in perfused rat kidney. Lead acetate caused a time and concentration-dependent increase in enzymuria. The effect of concurrent perfusion with lead and L-arginine (L-arg) or L-N(G)-nitro arginine methyl ester (L-NAME) [substrate and inhibitor of NO synthase respectively] in the perfusion fluid was also studied by measuring NAG activity in the perfusate kidney rat. L-arg (2 mM) has significantly decreased the lead-induced NAG release (P < 0.001), and L-NAME (0.1 mM) has significantly increased the lead-induced enzyme release in a time-dependent manner (P < 0.001). Moreover, histological studies using light microscope showed that some of the epithelial cells of the proximal convoluted tubules are degenerated or necrotic and desquamated into the lumens in rat treated with lead acetate. This change occurs at 50 microg/dl of lead acetate and was increased by addition of L-NAME to lead acetate. However, addition of L-arg had no effect on histology of lead nephrotoxicity. This may suggest that lead may interfere with the NO system in rat kidney.

Acetylglucosaminidase↗
PubMed1999

Ceramide induces cytochrome c release from isolated mitochondria.

This chapter addresses the role of mitochondria in apoptosis. Emphasis is put on the recently observed influence of ceramides on mitochondrial functions. We report here that N-acetylsphingosine (C2-ceramide), N-hexanoylsphingosine (C6-ceramide) and, to a much lesser extent, C2-dihydroceramide, induce cytochrome c (cyt c) release from isolated rat liver mitochondria. Ceramide-induced cyt c release is prevented by a low concentration of Bcl-2. The release takes place when cyt c is oxidized, but not when it is reduced. Upon cyt c release mitochondrial oxygen consumption, mitochondrial transmembrane potential (delta psi m) and Ca2+ retention are diminished. Bcl-2 prevents, and addition of cyt c reverses, the alteration of these mitochondrial functions. In ATP-energized mitochondria ceramides do not alter delta psi m, neither when cyt c is oxidized nor when it is reduced. This rules out a non-specific disturbance by ceramides of mitochondrial-membrane integrity. It is concluded that some of the apoptogenic properties of ceramides are mediated via their interaction with mitochondrial cyt c followed by its release.

Animals↗
PubMed1998

Endogenous nitric oxide modulates naloxone-precipitated withdrawal signs in a mouse model with acute cholestasis.

Cholestasis liver disease is associated with clinical and experimental findings consistent with increased opioidergic neuromodulation, increased plasma total activity, and elevated plasma enkephalin concentrations. The effect of the nitric oxide (NO) synthase inhibitor, L-nitro-arginine (L-NA, 0.03, 0.1, 0.3, 1 mg/kg), and the nitric oxide precursor, L-Arg (30 mg/kg), on antinociception induced by bile duct resection or sham operation, as well as on opioid dependence, was examined in male albino Swiss mice. Repeated (5 days) administration of L-NA attenuated signs of dependence, as assessed by naloxone (5 mg/kg)-precipitated withdrawal, and decreased the antinociception; however, L-Arg potentiated withdrawal signs and increased the antinociception. The results of this study support the involvement of the L-arginine/nitric oxide pathway in the opioidergic-dependent manifestation of cholestasis in an animal model.

Acute Disease↗
PubMed1997

Nitric oxide synthase activity in mitochondria.

In the present study we show the existence of a functional nitric oxide synthase (NOS) in rat liver mitochondria. The enzyme uses L-arginine (L-arg) to produce nitric oxide (NO) and L-citrulline, and is Ca2+-dependent. L-Arg analogues, N(omega)monomethyl-L-arg and N(omega)-nitro-L-arg, inhibit the enzyme, and D-arginine is not a substrate for it. We found mitochondrial NOS (mtNOS) activity associated with the inner mitochondrial membrane but not with the matrix fraction. In intact, succinate-energized mitochondria, the enzyme is constitutively active and exerts substantial control over mitochondrial respiration and membrane potential. The activity is further stimulated when Ca2+ is taken up by mitochondria. We suggest that the existence of mtNOS and its Ca2+ dependence are highly relevant for mitochondrial functioning.

Animals↗
PubMed1997

Inhibition by L-NA, a nitric oxide synthase inhibitor, of naloxone-precipitated withdrawal signs in a mouse model of cholestasis.

Following the naloxone administration in bile duct resected animals, striking opioid withdrawal signs are observed due to increased opioidergic tone. Pretreatment of animals with L-nitro arginine, a nitric oxide synthase inhibitor, reduces the naloxone-precipitated withdrawal signs as well as increase the antinociception. The results of this study support evidence for the involvement of the L-arg-nitric oxide pathway in opioidergic-dependent manifestation of cholestasis in an animal model.

Analgesia↗
PubMed1996

Inhibition by lithium and rubidium of gentamicin-induced release of N-acetyl-beta-D-glucosaminidase from perfused rat kidney.

N-acetyl-beta-D-glucosaminidase (NAG) is one of the sensitive hydrolytic lysosomal enzymes which is released after renal tubular damages. We studied gentamicin-induced nephrotoxicity by determining the NAG release in perfused rat kidney. 100 micrograms/ml of gentamicin caused a time-dependent increase in enzymuria, peaking at 90 min. At this time the released NAG is about sixfold more than the control. The effect of concurrent perfusion with 100 micrograms/ml gentamicin and with 0.5 mmol/l lithium chloride or 0.5 mmol/l rubidium chloride in the perfusion fluid was also studied by measuring NAG activity in the perfusate. Both cations decrease the gentamicin-induced NAG release. However, the inhibitory effect of lithium chloride may be due to interference of this ion with the polyphosphoinositide cycle in renal tubular lysosomal membranes. There is no obvious evidence for an inhibitory effect of rubidium chloride.

Acetylglucosaminidase↗
PubMed1996

Cyclosporin A-induced functional and morphological changes in pilocarpine treated rat submandibular glands.

The effects of long-term administration of Cyclosporin A (CSA), an immunosuppressive agent, on submandibular glands of male albino rats were investigated. Sialochemistry studies revealed a reduction of pilocarpine-stimulated flow rates to 54% compared to the controls. Salivary Mg(2+) and K+ were elevated and a marked decrease in total protein concentration was observed. Light and electron microscopic features of treated glands show marked changes at tissue level. An irregular pattern of the nucleus, mitochondrial alterations, reduction in the number of secretory granules and their aggregation, disturbances of cytoplasmic organelles, and isometric vacuolation were among the most striking findings. Our results show that CSA causes marked functional and morphological alterations in rat submandibular glands, which may be due to the drug's direct effects on the tissue.

Animals↗
PubMed1996

Effects of cyclosporine A on the functions of submandibular and parotid glands of rats.

1. The present study was designed to investigate the possible effects of long-term (45 days) administration of therapeutic doses of cyclosporine A (25 mg/kg/day), on the functions of submandibular and parotid glands of rats. Pure submandibular and parotid saliva were collected intraorally by microcannulation of the ducts. 2. The weight gains of the treated animals during the study and the weights of the salivary glands at the end of 45 days were reduced significantly as compared with those of controls. 3. Sialochemistry studies revealed a marked decrease in total protein concentration in saliva obtained from submandibular glands (P < 0.05). 4. Determination of electrolyte concentrations in saliva of submandibular gland and serum showed considerable differences between treated and control groups. 5. Significant elevation of amylase activity in serum and parotid saliva was observed in the treated rats in comparison with controls (P < 0.001). 6. Data presented here indicates that long-term administration of therapeutic doses of cyclosporine A causes significant alterations in salivary output and composition.

Amylases↗
PubMed1995

On the relation of calcium channel blockers to rat parotid and submandibular glands function in vivo.

1. The effects of nifedipine, verapamil and diltiazem on rat parotid and submandibular glands function were studied. 2. Nifedipine (5 mg/kg), verapamil (5 mg/kg) and diltiazem (10 mg/kg) were injected intraperitoneally 15 min before saliva collection. 3. Animals were anesthetized with 50 mg/kg of sodium pentobarbital and 8 mg/kg of pilocarpine was used as secretagogue. 4. Submandibular saliva was analyzed for flow rate, protein and calcium concentrations; and parotid saliva for calcium and amylase contents. 5. In treated groups, flow rate and calcium of submandibular saliva were significantly lower than controls. Parotid calcium in the nifedipine group was decreased and in verapamil and diltiazem groups was increased. Parotid amylase was significantly decreased in both the nifedipine and diltiazem groups. 6. It is concluded that a blockade of calcium channels in salivary glands acinar cells by CCBs causes some alterations in salivary secretions.

Amylases↗
PubMed1995

Effects of vinca alkaloids on rat parotid and submandibular glands function in vivo.

1. Vincristine (1 mg/kg) and vinblastine (2 mg/kg) were injected intraperitoneally into the rats, 24 hr before the experiments. 2. Animals were anesthetized with 50 mg/kg of sodium pentobarbital and saliva was collected from vincristine-treated, vinblastine-treated and control animals using 8 mg/kg of pilocarpine as secretagogue. 3. Parotid saliva was analyzed for protein, amylase and Ca2+ content, and submandibular saliva for flow rate, protein and Ca2+ concentration. 4. Saliva from two treated groups was significantly lower (P < 0.01) in flow rate, amylase and protein content than that of control group. Calcium level was significantly increased (P < 0.05) in treated animals. 5. It is concluded that the antisecretory effects of vinca alkaloids may be consistent with their actions on salivary cell microtubules.

Amylases↗