Cellular Biology |
From the Department of Molecular and Cellular Pharmacology, University of Miami Medical Center, Fla.
Correspondence to Keith A. Webster, Department of Molecular and Cellular Pharmacology, Rosenstiel Medical Science Building, Room 6038, University of Miami Medical Center, 1600 NW 10th Ave, Miami, FL 33136. E-mail kwebster{at}chroma.med.miami.edu
| Abstract |
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Key Words: ceramide JNK ischemia/reperfusion redox free radical
| Introduction |
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The contribution of free radicals to reperfusion damage is well documented, supported both by direct measurements and by the protective effects of antioxidants.1 5 8 9 Electron spin resonance spectroscopy indicates that reperfusion causes a burst of free radicals that peaks 2 to 4 minutes after reperfusion and continues for several hours.10 The source of the free radicals may be the mitochondrial electron transport chain, which is inhibited during ischemia and reactivated by reoxygenation.4 Direct intramyocyte targets for free radical oxidations include polyunsaturated fatty acids, membrane phospholipids, and protein sulfhydryl groups.1 11 12 The latter may include relatively specific targets such as myofilament proteins, ion transporters, and ß-adrenergic receptors.12 13 14 Indirect targets for free radicals include the mitogen-activated protein kinase (MAPK)/extracellular signalregulated kinase (ERK), p38, and stress-activated protein kinase (SAPK)/c-Jun N-terminal kinase (JNK) signaling cascades, all of which have been shown to be activated by oxidative stress in different models of ischemia and reperfusion.15 16 17 18 19 The intermediate signal(s) for the activation of MAPK/JNKs by oxidative stress has not been determined.
We report here that reoxygenation of hypoxic cardiac myocytes mediates a rapid activation of neutral sphingomyelinase (nSMase) and increased intracellular levels of ceramide. This was blocked by pretreatments with antioxidants and preceded the induction of JNK. The results indicate that nSMase activation with subsequent ceramide signaling may be one of the earliest responses of cardiac myocytes to oxidative stress in this model of ischemia-reperfusion.
| Materials and Methods |
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Ceramide Assays
Ceramide was quantified by a modified
sn-1,2-diacylglycerol (DAG) kinase
assay.23 Briefly, lipids were extracted from cell
pellets with chloroform/methanol, saponified,24 and
resuspended by sonication in a detergent mixture. Samples were
incubated for 2 hours in a reaction mix containing bacterial DAG kinase
and [
-32P]ATP.
[
-32P]-Ceramides were resolved by
silica gel, subjected to autoradiography, and
quantified by liquid scintillation counting. Ceramides type III and IV,
diacylglycerol, and monoacylglycerol, which were also
phosphorylated by DAG kinase, were used as
standards.
Sphingomyelinase Assays
Sphingomyelinase activities were measured according to previous
reports.25 26 Briefly, for nSMase, soluble protein cell
extracts were incubated in a reaction mix containing 9 nmol
sphingomyelin and 20 000 dpm
[N-methyl-14C]sphingomyelin per
sample for 30 minutes at 37°C. The
[14C]phosphocholine hydrolyzed by nSMase was
extracted from each sample with chloroform:methanol (2:1), and
H2O and quantified by scintillation counting. For
the measurement of acidic sphingomyelinase (aSMase) activity, the
reactions were conducted at pH 5.0 with 1 mmol/L EDTA instead of
7.4, as described previously.26 All activities were
normalized to total cell protein.
Kinase Assays
JNK and MAPK activities in cellular lysates were determined by
using recombinant glutathione S-transferasec-Jun (1141)
or myelin basic protein (MBP) exactly as described
previously.17 Immunoprecipitation of cleared cell
lysates with JNK1/JNK2-specific or ERK2-specific polyclonal antibodies
(Santa Cruz Biotechnology) was also exactly as described
previously.17 27 28 29 Immunocomplex kinase assays were
performed using 2 µg of purified glutathione
S-transferasec-Jun (1141) or 20 µg of MBP. ERK and JNK
reactions were resolved in 12.5% discontinuous
SDS-polyacrylamide gels, dried, and exposed to Kodak X-AR film
for autoradiography.
Reagents
Fumonisin B1 and bacterial DAG kinase were from Calbiochem; C2-
and C6-ceramides were from Biomol; ceramides type III and IV,
diacylglycerol, monoacylglycerol, NAC,
-phenyl
N-tert-butylnitrone (PBN), and MBP were from Sigma; and
[N-methyl-14C]sphingomyelin and
[
-32P]ATP were from Amersham Life
Science.
| Results |
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150% of
the basal aerobic level after 24 hours of hypoxia.
Reoxygenation resulted in a burst of ceramide
production, which was apparent within the first 3 minutes, and
peaked after 10 minutes, with 225% of the aerobic level. Ceramide
levels began to decline after 10 minutes, there was a second smaller
peak after 2 hours, and then the levels declined almost to basal after
4 hours. The early accumulation of ceramide is consistent with
a response to redox stress caused by reoxygenation.
Hydrogen peroxide (0.5 mmol/L) treatment also caused an elevation
of ceramide that peaked within 8 minutes at 240% of control and
declined rapidly to just above baseline after 30 minutes. Hydrogen
peroxide at 0.5 mmol/L was close to the optimal concentration,
given that there was less ceramide produced (and lower nSMase
activation) with 0.1 or 1.0 mmol/L (data not shown).
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Activation of Neutral but Not aSMase by Reoxygenation
Ceramide can be generated by a de novo pathway involving ceramide
synthase or by hydrolysis of sphingomyelin by sphingomyelinases.
Pretreatment of cultures with the ceramide synthase
inhibitor fumonisin B1 (1 to 10 µmol/L, previously
shown to be optimal for inhibiting ceramide synthase)30 31
did not decrease ceramide accumulation at any time point, suggesting
that de novo synthesis was not required (data not shown). There are 2
major sphingomyelinases in most cell types; aSMase is predominantly
lysosomal, and neutral Mg2+-dependent
sphingomyelinase (nSMase) is present in both membrane-bound and
cytosolic compartments.32 33 34 Figures 2A
and 2B
show the activities of nSMase
and aSMase in cardiac myocytes after exposure to hypoxia and
reoxygenation, as described in Figure 1
. nSMase
activity was
120% of the aerobic level after 24 hours of
hypoxia, and it increased abruptly to 150% of aerobic level 5
to 10 minutes after reoxygenation. The nSMase activity
remained high for 2 to 4 hours and then declined. There was a small but
reproducible dip in nSMase activity at 3 minutes; we do not know the
reason for this. H2O2 also
caused nSMase activation that peaked within 5 minutes at
150%
control and declined below baseline after 30 minutes. The results are
consistent with the ceramide results described in Figure 1
. There was no significant change in the activity of the aSMase
under any condition at any time (Figure 2B
). These results are
consistent with activated nSMase as the source of
ceramide accumulation in reoxygenated cardiac myocytes.
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Ceramide Accumulation and nSMase Are Quenched by NAC
Pretreatment
Previous studies in other cell types have shown that oxidative
stress can stimulate ceramide production.35 36 37 38 39 To
determine whether nSMase activation and ceramide accumulation were
related to increased oxidative stress during
reoxygenation, cardiac myocytes were treated with
50 mmol/L NAC 30 minutes before reoxygenation, and
both ceramide and nSMase were measured again as described in
Figures 1
and 2
. Figure 3
shows that NAC treatment reduced both ceramide generation and nSMase
activation at all time points by as much as 80%. The cumulative effect
of NAC treatment in reducing ceramide accumulation and nSMase activity
at all time points was significant (P<0.01). Interestingly,
NAC treatment did not significantly affect aerobic ceramide or nSMase
activities.
|
Activation of JNK by Ceramide
We and others15 17 have shown that hypoxia
and reoxygenation of cardiac myocytes induces JNK.
Ceramide has been shown previously to induce JNK in other
cells.40 To determine whether ceramide generation after
reoxygenation may be involved with JNK activation, we
compared the kinetics of JNK activation and ceramide accumulation at
early and late time points after reoxygenation. As
shown in Figure 4
, there was a close
correlation; ceramide was elevated slightly earlier than JNK (within 1
minute), and both activities peaked after 10 to 15 minutes and declined
thereafter. Both remained elevated for 2 to 4 hours after
reoxygenation. Therefore, the kinetics of ceramide
accumulation is compatible with a role for ceramide in the activation
of JNK.
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There are no specific inhibitors of nSMase and no agents
that specifically block ceramide signaling; therefore, it is not
possible to determine directly whether JNK activation by
reoxygenation is dependent on ceramide accumulation. If
elevated intracellular ceramide is a signal for JNK activation during
reoxygenation, then other treatments that increase
intramyocyte ceramide should also activate JNK. Therefore, we
asked whether treatment with extracellular ceramides activated
JNK. Figure 4C
shows the responses to treatments with the
cell-permeant C2- and C6-ceramides and with natural ceramides (types
III and IV). With all ceramide treatments, there was a lag before JNK
was activated; this is probably because of the time required
for equilibration of the extracellular ceramides, which have poor
aqueous solubility and are delivered as DMSO-BSA complexes.
Consistent with this, the cell-permeant ceramides (100
µmol/L) activated JNK within 20 minutes, and there was a
longer lag time with the less soluble natural ceramides (30
µmol/L). Ceramides did not affect ERK activity (see third and fifth
panels in Figure 4C
). These results indicate that extracellular
ceramides can initiate a pathway for JNK activation in cardiac myocytes
and are compatible with a role for ceramide in the
reoxygenation-mediated activation of JNK.
Ceramide Accumulation and JNK Are Quenched in Parallel by
Antioxidant Pretreatment
Cardiac myocytes were cultured as described in Figure 3
and
were pretreated with the indicated antioxidant before
reoxygenation, as described for NAC. Cells were
harvested at the indicated times for ceramide assays or after 30
minutes for JNK assay as described above. Pretreatment with either PBN
(10 mmol/L) or ß-mercaptoethanol (ß-ME; 1 mmol/L) reduced
reoxygenation-mediated ceramide increases at all time
points, although the inhibition was not complete (Figure 5A
). The cumulative effect of
antioxidants on the suppression of ceramide production was
significant for PBN and ß-ME (P<0.05). Figure 5B
shows the effects of antioxidant pretreatment on JNK
activity. Again, all treatments reduced
reoxygenation-mediated JNK activity by >50%
(determined by densitometric analysis of the autoradiograph,
not shown).
|
Extracellular Ceramides but Not Reoxygenation
Induce JNK in Nonmuscle Fibroblasts
We previously reported that JNK was not induced by
reoxygenation of nonmuscle cardiac fibroblasts (NMFs),
indicating that JNK activation was at least partially specific for
cardiac myocytes.17 To determine whether the absence of
JNK activation was associated with a corresponding absence of ceramide
accumulation in these cells, we measured and correlated nSMase,
ceramide, JNK, and ERK activities after hypoxia and
reoxygenation. These results are shown in Figure 6
. Figure 6A
shows that although
ceramide accumulated in NMFs during exposure to hypoxia, there
was no subsequent increase after reoxygenation.
Therefore, either reoxygenation is less stressful to
NMFs or the signaling pathways activated by free radicals are
different. This is supported by nSMase measurements in Figure 6B
. In contrast to cardiac myocytes, there was no increase of
nSMase activity after reoxygenation of NMFs. Figure 6C
confirms that JNK was not activated by
reoxygenation in NMFs at any time. To confirm that NMFs
were responsive to extracellular ceramides, we measured JNK after
ceramide addition, as described in Figure 4
. JNK was strongly
induced in NMFs by all ceramide treatments (Figure 6D
).
Therefore, the absence of JNK activation by
reoxygenation of NMFs correlates with the absence of
nSMase activation and ceramide accumulation in these cells.
Similarly to the results with cardiac myocytes, ERK activity was not
significantly affected by ceramide treatments of NMFs (Figure 6E
).
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| Discussion |
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This is the first study to demonstrate that hypoxia and
reoxygenation activate nSMase and cause
ceramide accumulation. Activation of this pathway has been described in
other cell types treated with cytokines (tumor necrosis factor
[TNF]
), ionizing radiation, and
H2O2 (reviewed in Reference
3232 ). Therefore, nSMase appears to be activated by multiple
stimuli associated with oxidative stress. The molecular mechanisms for
this regulation of nSMase activity are not understood; in the case of
stimulation with TNF-
, nSMase may be activated directly by
binding to the TNF-
receptor45 ; other studies indicate
that nSMase is negatively regulated by glutathione.46 47 48
In our studies, nSMase was activated in 2 steps, first, during
chronic hypoxia, and second, immediately after
reoxygenation. Glutathione levels fall precipitously
during hypoxia17 48 ; therefore, the
hypoxia phase of nSMase activation may involve release of
glutathione inhibition. The rapid kinetics of nSMase activation during
reoxygenation, as well as the responses to
antioxidants, is consistent with an upstream role of oxygen
free radicals as the initiators. The mechanism of this initiation is
not known, but presumably it could involve positive redox regulation of
nSMase or negative regulation of an inhibitor.
Previous studies have shown that ceramide can activate JNK through pathways involving TGF-ßactivated kinase-1 or RAS.28 32 40 In our studies, the first peak of ceramide accumulation after reoxygenation coincided with the initiation of JNK activity. Activated JNK was detected 8 minutes after reoxygenation and peaked at 15 minutes. JNK activity remained elevated over the basal activity for at least 4 hours. The correlation of ceramide accumulation with JNK activation is consistent with a possible cause-and-effect relationship. Smaller inductions of ceramide than those reported here have been shown to strongly activate JNK and apoptosis in other cell types.49 Such a functional relationship between ceramide accumulation and JNK activation is supported by the observation that treatments with cell-permeant and natural ceramides activated JNK in cardiac myocytes. nSMase activity, ceramide accumulation, and JNK activation were all strongly inhibited by NAC and other antioxidants, supporting a common pathway, and neither nSMase nor JNK was induced in reoxygenated nonmuscle fibroblasts. JNK was not strongly induced by hypoxia alone in either cardiac myocytes or NMFs, even though the ceramide level increased. The reason for this is not clear, but it may be related to the more gradual nature of ceramide accumulation during hypoxia, the site of production and compartmentalization of ceramide,50 and the acute nature of JNK activation.51 Alternatively, there may be a moderate, barely detectable but chronic activation of JNK during hypoxia, which would be consistent with the elevated levels of c-Jun seen under these conditions.21 Therefore, although our data support a role for ceramide in the activation of JNK by reoxygenation (and this is consistent with previous results), confirmation of such a role will require further experiments. Furthermore, increased ceramide may not be the only stimulus to activate JNK under these conditions. Another pathway has been proposed involving the oxidative inactivation of specific JNK phosphatases.52 Although the involvement of this pathway is also speculative, such activity may contribute to the activation of JNK by oxidative stress described here.
H2O2 at a concentration of 0.5 mmol/L quantitatively mimicked the effects of reoxygenation. Both nSMase activity and ceramide accumulation peaked early, within 10 minutes of H2O2 treatment, and then declined. The relatively longer-lasting effects of hypoxia-reoxygenation compared with H2O2 may be related to the reduced levels of glutathione, caused by hypoxia, that only recover gradually after reoxygenation, and as a consequence, may prolong the oxidative stress.17 Despite their similar effects on nSMase and ceramide, reoxygenation and H2O2 may have differential effects on MAPKs. H2O2 was previously reported to induce all MAPKs in cardiac myocytes, as well as other cells, with an early activation of p38 followed by slightly later activation of ERK and JNK/SAPK.18 52 53 54 55 Hypoxia-reoxygenation, on the other hand, causes strong activation of JNK, but modest or no activation of ERK and p38.17 The differences may be related to the different sites and species of free radicals generated by these stimuli.32 56 Ceramide has also been reported to preferentially induce JNK over ERK in other cells,57 and reperfusion causes preferential activation of JNK over ERK in whole-heart preparations.18 Although these observations are consistent with ceramide-mediated activation of JNK by redox stress, other pathways must also be involved, because H2O2, but not ceramide, activates ERK and p38.
Ceramide is the central molecule in the highly conserved sphingomyelin
signaling system and serves as a second messenger for numerous cellular
functions ranging from proliferation and differentiation to growth
arrest and apoptosis (reviewed in References 32 and 5632 56 ).
Increased tissue ceramide has been reported in reperfused rat
hearts.58 Alterations of ceramide and JNK activities have
been associated with ischemia and reperfusion of both kidney
and heart, and may be involved in apoptosis
pathways.58 59 60 61 Previous studies have shown ceramide
accumulation at late stages of apoptosis in cardiac myocytes
subjected to hypoxia and metabolic
inhibition.58 Other studies suggest that the sites of
generation and cellular compartmentation determine cellular responses
to ceramide changes50 and that increased aSMase activity
is linked to apoptosis, whereas nSMase activation favors cell
survival.62 63 Under the conditions of
hypoxia-reoxygenation described here,
30%
of the cardiac myocytes die by apoptosis after 16 to 20 hours
of reoxygenation, whereas 70%
survive.17 64 The contributions of nSMase activation,
increased ceramide generation, and JNK activity to these events are not
known.
| Acknowledgments |
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Received August 23, 1999; accepted October 27, 1999.
| References |
|---|
|
|
|---|
2. Gottlieb RA, Burleson KO, Kloner RA, Babior BM, Engler RL. Reperfusion injury induces apoptosis in rabbit cardiomyocytes. J Clin Invest. 1994;94:16121628.
3.
Ohno M, Takemura G, Ohno A, Misao J, Hayakawa Y,
Minatoguchi S, Fujiwara T, Fujiwara H. Apoptotic myocytes in
infarct area in rabbit hearts may be oncotic myocytes with DNA
fragmentation: analysis by immunogold electron microscopy
combined with in situ nick end-labeling. Circulation. 1998;98:14221430.
4. Kloner RA, Bolli R, Marbán E, Reinlib L, Braunwald E. Medical and cellular implications of stunning, hibernation, and preconditioning: an NHLBI workshop. Circulation. 1998;1848:18671860.
5.
Horwitz LD, Fennessey PV, Shikes RH, Kong Y. Marked
reduction in myocardial infarct size due to prolonged infusion of an
antioxidant during reperfusion. Circulation. 1994;89:17921801.
6. Kusuoka H, Marbán E. Cellular mechanisms of myocardial stunning. Annu Rev Physiol. 1992;54:243256.[Medline] [Order article via Infotrieve]
7. Kajstura J, Cheng W, Reiss K, Clark WA, Sonnenblick EH, Krajewski S, Reed JC, Olivetti G, Anversa P. Apoptotic and necrotic myocyte cell death are independent contributing variables of infarct size. Lab Invest. 1996;74:86107.[Medline] [Order article via Infotrieve]
8. Grech ED, Bellamy CM, Jackson MJ, Muirhead RA, Faragher EB, Ramsdale DR. Free-radical activity after primary coronary angioplasty in acute myocardial infarction. Am Heart J. 1994;127:14431449.[Medline] [Order article via Infotrieve]
9. Grech ED, Dodd NJF, Jackson MJ, Morrison WL, Faragher EB, Ramsdale DR. Evidence for free radical generation after primary percutaneous transluminal coronary angioplasty recanalization in acute myocardial infarction. Am J Cardiol. 1996;77:122127.[Medline] [Order article via Infotrieve]
10. Bolli R, Patel BS, Jeroudi MO, Lai EK, McCay PB. Demonstration of free radical generation in stunned myocardium of intact dogs with the use of spin trap alpha-phenyl N-tert-butyl nitrone. J Clin Invest. 1988;82:476485.
11. Paradies G, Petrosillo G, Pistolese M, Di Venosa N, Serena D, Ruggiero FM. Lipid peroxidation and alterations to oxidative metabolism in mitochondria isolated from rat heart subjected to ischemia and reperfusion. Free Radic Biol Med. 1999;27:4250.[Medline] [Order article via Infotrieve]
12.
Chiamvimonvat N, ORourke B, Kamp TJ, Kallen RG,
Hofmann F, Flockerzi V, Marbán E. Functional consequences of
sulfhydryl modification in the pore-forming subunits of
cardiovascular Ca2+ and
Na+ channels. Circ Res. 1995;76:325334.
13. Gao WD, Atar D, Liu Y, Perez NG, Murphy AM, Marbán E. Role of troponin I proteolysis in the pathogenesis of stunned myocardium. Circ Res. 1997;80:393399.
14. Bolli R. Basic and clinical aspects of myocardial stunning. Prog Cardiovasc Dis. 1998;40:477516.[Medline] [Order article via Infotrieve]
15.
Seko Y, Tobe K, Ueki K, Kadowaki T, Yazaki Y.
Hypoxia and hypoxia/reoxygenation
activate raf-1, mitogen-activated protein kinase
kinase, mitogen-activated protein kinase, and S6 kinase in
cultured rat cardiac myocytes. Circ Res. 1996;78:8290.
16. Knight RJ, Buxton DB. Stimulation of c-Jun kinase and mitogen-activated protein kinase by ischemia and reperfusion in the perfused rat heart. Biochem Biophys Res Commun. 1996;218:8388.[Medline] [Order article via Infotrieve]
17.
Laderoute KR, Webster KA.
Hypoxia/reoxygenation stimulates Jun kinase
activity through redox signaling in cardiac myocytes. Circ
Res. 1997;80:336344.
18.
Clerk A, Fuller SJ, Michael A, Sugden PH. Stimulation
of stress-regulated mitogen-activated protein kinases
(stress-activated protein kinases/c-Jun-N-terminal kinases and
p38-mitogen-activated protein kinases) in perfused rat hearts
by oxidative and other stresses. J Biol Chem. 1998;273:72287234.
19.
Sugden PH, Clerk A. Stress-responsive
mitogen-activated protein kinases in the
myocardium. Circ Res. 1998;83:345352.
20. Webster KA, Bishopric NH. Molecular regulation of cardiac myocyte adaptations to chronic hypoxia. J Mol Cell Cardiol. 1992;24:741751.[Medline] [Order article via Infotrieve]
21.
Webster KA, Discher D, Bishopric NH. Induction and
nuclear accumulation of Fos and Jun proto-oncogenes in hypoxic cardiac
myocytes. J Biol Chem. 1993;268:1685216859.
22.
Bishopric NH, Simpson PC, Ordahl CP. Induction of the
skeletal actin gene in
1-adrenoceptor mediated
hypertrophy of rat cardiac myocytes. J Clin
Invest. 1987;80:11941199.
23.
Preiss J, Loomis CR, Bishop WR, Stein R, Niedel JE,
Bell RM. Quantitative measurement of sn-1,2-diacylglycerols
present in platelets, hepatocytes, and
ras- and sis-transformed normal rat kidney cells.
J Biol Chem. 1986;261:85978600.
24.
Dressler KA, Kolesnick RN. Ceramide-1-phosphate, a
novel phospholipid in human leukemia (HL-60) cells. J Biol
Chem. 1990;265:1491714921.
25. Quintern LE, Sandhoff K. Human acid sphingomyelinase from human urine. Methods Enzymol. 1991;197:536540.[Medline] [Order article via Infotrieve]
26. Wiegmann K, Schutze S, Machleidt T, Witte D, Kronke M. Functional dichotomy of neutral and acidic sphingomyelinases in tumor necrosis factor signaling. Cell. 1994;78:10051015.[Medline] [Order article via Infotrieve]
27.
Coso OA, Chiariello M, Kalinec G, Kyriakis JM, Woodgett
J, Gutkind JS. Transforming G protein-coupled receptors potently
activate JNK (SAPK). J Biol Chem. 1995;270:56205624.
28.
Bagrodia S, Derijard B, Davis RJ, Cerione RA. Cdc42 and
PAK-mediated signaling leads to Jun kinase and p38
mitogen-activated protein kinase activation. J Biol
Chem. 1995;270:2799527998.
29. Kyriakis JM, Banerjee P, Nikolakaki E, Dai T, Rubie EA, Ahmad MF, Avruch J, Woodgett JR. The stress-activated protein kinase subfamily of c-Jun kinases. Nature. 1994;369:156160.[Medline] [Order article via Infotrieve]
30.
Wang E, Norred WP, Bacon CW, Riley RT, Merrill AH Jr.
Inhibition of sphingolipid biosynthesis by fumonisins. J
Biol Chem. 1998;266:1448614490.
31.
Merrill AH Jr, van Echten G, Wang E, Sandhoff K.
Fumonisin B1 inhibits sphingosine (sphinganine)
N-acyltransferase and de novo sphingolipid biosynthesis in
cultured neurons in situ. J Biol Chem. 1993;268:2729927306.
32. Mathias S, Pena LA, Kolesnick RN. Signal transduction of stress via ceramide. Biochem J. 1998;335:465480.
33.
Liu B, Hassler DF, Smith GK, Weaver K, Hannun YA.
Purification and characterization of a membrane bound neutral pH
optimum magnesium-dependent and
phosphatidylserine-stimulated sphingomyelinase from
rat brain. J Biol Chem. 1998;273:3447234479.
34.
Okazaki T, Bielawska A, Domae N, Bell RM, Hannun YA.
Characteristics and partial purification of a novel cytosolic
magnesium-independent, neutral sphingomyelinase activated in
the early signal transduction of 1
-25-dihydroxyvitamin
D3-induced HL-60 cell differentiation.
J Biol Chem. 1994;269:40704077.
35. Zager RA, Conrad S, Lochhead K, Sweeney EA, Igarashi Y, Burkhart KM. Altered sphingomyelinase and ceramide expression in the setting of ischemic and nephrotoxic acute renal failure. Kidney Int. 1998;53:573582.[Medline] [Order article via Infotrieve]
36.
Singh I, Pahan K, Khan M, Singh AK.
Cytokine-mediated induction of ceramide production is
redox-sensitive. J Biol Chem. 1998;273:2035420362.
37.
Bhunia AK, Han H, Snowden A, Chatterjee S.
Redox-regulated signaling by lactosylceramide in the proliferation of
human aortic smooth muscle cells. J Biol Chem. 1997;272:1564215649.
38. Goldkorn T, Balaban N, Shannon M, Chea V, Matsukuma K, Gilchrist D, Wang H, Chan C. H2O2 acts on cellular membranes to generate ceramide signaling and initiate apoptosis in tracheobronchial epithelial cells. J Cell Sci. 1998;111:32093220.[Abstract]
39. Singh I, Pahan K, Khan M, Singh A. Cytokine-mediated induction of ceramide production is redox-sensitive. J Biol Chem. 1998;273:2035420362.
40.
Shiradabe K, Yamaguchi K, Shibuya H, Irie K, Matsuda S,
Moriguchi T, Gotoch Y, Matsumoto K, Nishida E. TAK1 mediates the
ceramide signaling to stress-activated protein kinase/c-Jun
N-terminal kinase. J Biol Chem. 1997;272:81418144.
41.
Jaffrezou JP, Maestre N, Mas-Mansat VD, Bezombes C,
Levade T, Laurant G. Positive feedback control of neutral
sphingomyelinase activity by ceramide. FASEB J. 1998;12:9991006.
42.
Gudz TI, Tserng K, Hoppel CL. Direct inhibition of
mitochondrial respiratory chain complex III by cell-permeable ceramide.
J Biol Chem. 1997;272:2415424158.
43.
Ghafourifar P, Klein SD, Schucht O, Schenk U, Pruschy
M, Rocha S, Richter C. Ceramide induces cytochrome c release from
isolated mitochondria. J Biol Chem. 1999;274:60806084.
44.
Quillet-Mary A, Jaffrezou JP, Mansat V, Bordier C,
Naval J, Laurent G. Implication of mitochondrial hydrogen peroxide
generation in ceramide-induced apoptosis. J Biol
Chem. 1997;272:2138821395.
45. Adam-Klages S, Adam D, Wiegmann K, Struve S, Kolanus W, Schneider-Mergener K, Kronke M. FAN, A novel WD-repeat protein, couples the p55 TNF-receptor to neutral sphingomyelinase. Cell. 1996;86:937947.[Medline] [Order article via Infotrieve]
46.
Liu B, Andrieu-Abadie N, Levade T, Zhang P, Obeid LM,
Hannun YA. Glutathione regulation of neutral sphingomyelinase in tumor
necrosis factor-
-induced cell death. J Biol
Chem. 1998;273:1131311320.
47.
Liu B, Hannun YA. Inhibition of the neutral
magnesium-dependent sphingomyelinase by glutathione. J Biol
Chem. 1997;272:1628116287.
48. Yoshimura S, Banno Y, Nakashima S, Hayashi K, Yamakawa H, Sawada M, Sakai N, Nozawa Y. Inhibition of neutral sphingomyelinase activation and ceramide formation by glutathione in hypoxic PC12 cell death. J Neurochem. 1999;73:675683.[Medline] [Order article via Infotrieve]
49. Verheij M, Bose R, Lin XH, Yao B, Jarvis WD, Grant S, Birrer MJ, Szabo E, Zon LI, Kyriakis JM, Haimovitz-Friedman A, Fuks Z, Kolesnick RN. Requirement for ceramide-initiated SAPK/JNK signaling in stress-induced apoptosis. Nature. 1996;380:7579.[Medline] [Order article via Infotrieve]
50.
Zhang P, Liu B, Jenkins GM, Hannun YA, Obeid LM.
Expression of neutral sphingomyelinase identifies a distinct pool of
sphingomyelin involved in apoptosis. J Biol
Chem. 1997;272:96099612.
51. Yao A, Takahashi T, Aoyagi T, Kinugawa K, Kohmoto O, Sugiura S, Serizawa T. Immediate-early gene induction and MAP kinase activation during recovery from metabolic inhibition in cultured cardiac myocytes. J Clin Invest. 1995;1:6977.
52. Clerk A, Michael A, Sugden PH. Stimulation of multiple mitogen-activated protein kinase sub-families by oxidative stress and phosphorylation of the small heat shock protein, HSP25/27, in neonatal ventricular myocytes. Biochem J. 1998;333:581589.
53.
Guyton KZ, Liu Y, Gorospe M, Xu Q, Holbrook NJ.
Activation of mitogen-activated protein kinase by
H2O2: role in cell survival
following oxidant injury. J Biol Chem. 1996;271:41384142.
54.
Bogoyevitch MA, Gillespie-Brown J, Ketterman AJ, Fuller
SJ, Ben-Levy R, Ashworth A, Marshall CJ, Sugden PH. Stimulation of the
stress-activated mitogen-activated protein kinase
subfamilies in perfused heart: p38/ERK mitogen-activated
protein kinases and c-Jun N-terminal kinases are activated by
ischemia/reperfusion. Circ Res. 1996;79:162173.
55. Aikawa R, Komuro I, Tsutomo Y, Zou Y, Kudoh S, Tanaka M, Shiojima I, Hiroi Y, Yazaki Y. Oxidative stress activates extracellular signal-regulated kinases through Src and Ras in cultured cardiac myocytes of neonatal rats. J Clin Invest. 1997;100:18131821.[Medline] [Order article via Infotrieve]
56. Hofmann K, Dixit VM. Ceramide in apoptosis: does it really matter? Trends Biochem Sci. 1998;23:374377.[Medline] [Order article via Infotrieve]
57. Coroneos E, Wang Y, Panuska JR, Templeton DJ, Kester M. Sphingolipid metabolites differentially regulate extracellular signal-regulated kinase and stress-activated protein kinase cascades. Biochem J. 1996;316:1317.
58. Bielawska AE, Shapiro JP, Jiang L, Melkonyan HS, Piot C, Wolfe CL, Tomei LD, Hannun YA, Umansky SR. Ceramide is involved in triggering of cardiomyocyte apoptosis induced by ischemia and reperfusion. Am J Pathol. 1997;151:12571263.[Abstract]
59. Zager RA, Iwata M, Conrad DS, Burkhart KM, Igarashi Y. Altered ceramide and sphingosine expression during the induction phase of ischemic acute renal failure. Kidney Int. 1997;52:6070.[Medline] [Order article via Infotrieve]
60. Ueda N, Kaushal GP, Hong X, Shah SV. Role of enhanced ceramide generation in DNA damage and cell death in chemical hypoxic injury to LLC-PK1 cells. Kidney Int. 1998;54:399406.[Medline] [Order article via Infotrieve]
61.
Yin T, Sandhu G, Wolfgang CD, Burrier A, Webb RL, Rigel
DF, Hai T, Whelan J. Tissue-specific pattern of stress kinase
activation in ischemic/reperfused heart and kidney.
J Biol Chem. 1997;272:1994319950.
62.
Riboni L, Prinetti A, Bassi R, Caminiti A, Tettamani G.
A mediator role of ceramide in the regulation of neuroblastoma Neuro2a
cell differentiation. J Biol Chem. 1995;270:2686826875.
63.
Auge N, Escargueil-Blanc I, Lajoie-Mazenc I, Suc I,
Andrieu-Abadie N, Pieraggi MT, Chatelut M, Thiers JC, Jaffrezou JP,
Laurent G, Levade T, Negre-Salvayre A, Salvaryre A. Potential role for
ceramide in mitogen-activated protein kinase activation and
proliferation of vascular smooth muscle cells induced by oxidized low
density lipoprotein. J Biol Chem. 1998;273:1289312900.
64. Webster KA, Discher DJ, Kaiser S, Hernandez O, Sato B, Bishopric NH. Hypoxia-activated apoptosis of cardiac myocytes requires reoxygenation or a pH shift and is independent of p53. J Clin Invest. 1999;104:239252.[Medline] [Order article via Infotrieve]
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