ToxSci Advance Access originally published online on March 30, 2006
Toxicological Sciences 2006 91(2):643-650; doi:10.1093/toxsci/kfj175
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Mitochondrial Thioredoxin-2 Has a Key Role in Determining Tumor Necrosis Factor-
–Induced Reactive Oxygen Species Generation, NF-
B Activation, and Apoptosis
* Division of Pulmonary, Asthma, Cystic Fibrosis and Sleep, Department of Pediatrics and Division of Pulmonary, Allergy and Critical Care Medicine, Department of Medicine, School of Medicine, Emory University, Atlanta, Georgia 30322
1 To whom correspondence should be addressed at Division of Pulmonary, Allergy and Critical Care Medicine, Department of Pediatrics, 2015 Uppergate Drive, Suite 350, Atlanta, GA 30322. E-mail: jhansen{at}emory.edu.
Received January 12, 2006; accepted March 1, 2006
 |
ABSTRACT |
|---|
 TOP ABSTRACT INTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
Tumor necrosis factor-
(TNF-) is a cytokine that is involved in numerous pathologies, in part through stimulation of the mitochondrial production of reactive oxygen species (ROS). Previous studies show that in addition to mitochondrial superoxide dismutase- and glutathione-dependent systems, mitochondria also contain thioredoxin-2 (Trx2), an antioxidant protein that can detoxify ROS. The purpose of this study was to determine whether Trx2 protects against oxidative damage triggered by TNF-. After a 30-min treatment in HeLa cells, TNF- (5–40 ng/ml) oxidized Trx2 but not cytoplasmic Trx1. Preferential, significant Trx2 oxidation occurred within 10 min of TNF- treatment. Moreover, overexpression of Trx2, but not Trx1, decreased TNF-–induced ROS generation, suggesting mitochondrial compartmentation of ROS production and subsequent specific detoxification by Trx2, not Trx1. Overexpression of Trx2 or the active-site mutant C93S Trx2 was used to evaluate their downstream effects following TNF- stimulation. Results showed that nuclear translocation of NF-
B was inhibited with Trx2 overexpression but not with the dominant negative active-site mutant C93S Trx2. Moreover, when cotransfected with a NF-B-luciferase reporter and then treated with TNF-, NF-B activity was significantly attenuated with Trx2 overexpression but not with C93S Trx2 expression. Trx2 overexpression, but not C93S Trx2, significantly inhibited TNF-–induced apoptosis as measured by terminal dUTP nick-end labeling assay. These findings support the interpretation that mitochondrial-generated ROS is a principal component in TNF-–induced effects and that Trx2 blocks TNF-–induced ROS generation and downstream NF-B activation and apoptosis.
Key Words: NF-B; apoptosis; oxidative damage; ROS generation; thioredoxin; TNF-.
|
INTRODUCTION |
|---|
|
TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
Tumor necrosis factor-
(TNF-) is a cytokine that is involved in multiple physiological functions, such as hematopoeisis and tumor regression (Carswell et al., 1975
), and disease states, such as Crohn's disease (Suenaert et al., 2002), pulmonary fibrosis (Miyazaki et al., 1995), asthma (Cembrzynska-Nowak et al., 1993), rheumatoid arthritis (Smith and Haynes, 2002), and diabetes (Hotamisligil et al., 1996). While the exact mechanisms of TNF-–induced effects are not completely understood, the production of reactive oxygen species (ROS) during TNF- signaling appears to be critical to propagate TNF- signaling. Multiple studies show that TNF-–induced ROS generation occurs primarily in the mitochondria (Goossens et al., 1995; Higuchi et al., 1998; Schulze-Osthoff et al., 1992; Shoji et al., 1995), suggesting that this organelle may be important in regulating downstream effects of TNF-, such as NF-B activation and apoptosis.
Multiple redox couples function in cells to detoxify ROS and regulate redox-sensitive processes (Jones et al., 2004). The most widely studied of these redox couples are the amino acid cysteine, the tripeptide glutathione (GSH), and the protein thioredoxin (Trx). Recent evidence has shown that these couples are independently regulated (Hansen et al., 2004; Nkabyo et al., 2002). Independent regulation of these couples allows for the selective, specific regulation of redox signaling and has been shown in various models systems (Hansen et al., 2004; Harper et al., 2001; Merwin et al., 2002; Trotter and Grant, 2003; Watson et al., 2003a). However, redox signaling is not only regulated by these different redox couples but may also be a consequence of redox differences between intracellular compartments and environments (Hansen et al., 2005). Past studies have focused on GSH compartmentation, but results have been variable and contradictory (Bellomo et al., 1992; Cotgreave, 2003; Voehringer et al., 1998). Recent studies have focused on Trx, which can overcome many problems found with organellar GSH measurement. Epidermal growth factor (EGF) is known to produce ROS upon binding to its receptor (Sundaresan et al., 1995). EGF stimulation of keratinocytes caused significant, specific cytosolic Trx oxidation, but mitochondrial and nuclear Trx pools were unaffected. In HeLa cells, nuclear Trx1 is much more reduced than cytoplasmic Trx1 pools, and upon treatment with tert-butyl hydroperoxide, a + 60-mV oxidation of Trx1 in the nucleus was observed, but a similar treatment produced only a + 35-mV oxidation of Trx1 in the cytosol, demonstrating compartmental differences in baseline redox states and sensitivity to oxidative stress (Watson and Jones, 2003). These studies provide a background for compartmentation of redox control, but few studies have examined regulation of mitochondrial redox signaling.
Trx systems exist in both the cytoplasm/nucleus and the mitochondria but are distinct. The Trx system in the mitochondria is completely independent of the cytosolic system, containing a unique gene product, Trx2 rather than Trx1, and a corresponding thioredoxin reductase (TR), TR2 rather than TR1. Furthermore, peroxiredoxin 3 (Prx3), a peroxidase using Trx2 as an electron donor, is present only in the mitochondria. Whereas GSH is in constant flux in and out of the mitochondria and interacts with the cytosolic GSH pool, the TR2/Trx2/Prx3 system is inclusive within the mitochondria and independent of the TR1/Trx1/Prx cytosolic system.
Although evidence is emerging to illustrate compartmentation of redox signaling, specific oxidation of Trx2 in response to physiologic signaling, such as TNF- stimulation, has not been demonstrated. Since ROS generation occurs within the mitochondria and is involved in NF-B activation and apoptosis, Trx2 is a likely candidate to regulate ROS production and TNF- signaling. In the present study, we utilize redox Western blotting techniques to show specific mitochondrial and cytosolic effects of TNF-. Our findings show that TNF- caused selective mitochondrial oxidation, as evidenced by oxidation of Trx2 but not Trx1. Moreover, overexpression of Trx2 inhibited TNF-–induced ROS generation and subsequent ROS-dependent downstream events, such as NF-B activation and apoptosis. These results show that mitochondrial Trx2 detoxifies TNF-–induced mitochondrial ROS production and, therefore, inhibits subsequent signaling events.
|
MATERIALS AND METHODS |
|---|
|
TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
Cell culture.
HeLa cells were purchased from American Tissue Culture Collection (Manassas, VA) and were grown in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum and antibiotics (penicillin/streptomycin). Cultures were maintained in a humidified incubator in a 5% CO2 atmosphere at 37°C and treated upon reaching 80–90% confluence. HeLa cells were chosen due to their high transfection efficiency as well as their well-documented use in TNF-
experiments by others. To best evaluate TNF- stimulation in the context of Trx2, treatment concentrations ranged from 0 to 40 ng/ml.
ROS generation by TNF-.
Cellular ROS generation was determined with 5-(and-6)-carboxy-2',7'-dichlorodihydrofluorescein diacetate (DCF) (Invitrogen, Carlsbad, CA) by incubating cells in DMEM with 1% fetal bovine serum and 100µM DCF for 30 min at 37°C (Wang and Joseph, 1999). An increase in DCF fluorescence is primarily due to the reaction with intracellular hydrogen peroxide, as noted by the manufacturer. Medium was replaced with Krebs-Ringer 4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid buffer containing various concentrations of TNF- (0–40 ng/ml) (Sigma, St Louis, MO). Changes in sample fluorescence were followed kinetically with a Molecular Devices M2 fluorescence microplate reader (Sunnyvale, CA) at 488-nm excitation and 520-nm emission for 30 min while maintaining at 37°C.
MitoSox Red mitochondrial superoxide indicator dye (Invirogen) was used to determine ROS in the mitochondria. Cells were treated with TNF- (0–40 ng/ml) for 30 min after which they were loaded with 5µM MitoSox Red in Hank's balanced salt solution for 10 min at 37°C. Cells were washed with warm buffer. MitoSox Red fluorescent intensity was determined at 510-nm excitation and 580-nm emission.
Trx redox Western analysis.
Trx1 redox state analysis was performed as previously described (Watson et al., 2003b). Separation of the oxidized and reduced forms of Trx1 was performed via Western blotting by native, nondenaturing polyacrylamide gel electrophoresis (15%). Proteins were electroblotted onto a nitrocellulose membrane prior to the immunodetection by a goat primary antibody for Trx1 (American Diagnostica, Greenwich, CT) and a donkey anti-goat AlexaFluor 680 secondary antibody (Molecular Probes, Carlsbad, CA). Membranes were scanned using the Odyssey Scanning system (Li-Cor, Lincoln, NE). Densitometric analysis of membranes was performed with the Odyssey Scanning software. Band densitometric values were used with the Nernst equation to estimate steady-state redox potential of Trx1 (Watson et al., 2003b).
Redox state of Trx2 was determine by the redox Western analysis with 4-acetamido-4'-maleimidylstilbene-2,2'-disulfonic acid as described by Halvey et al. (2005) based on the original method of Damdimopoulos et al. (2002).
Trx2 and C93S plasmids.
Human Trx2 was cloned in to pCDNA3.1 as performed by Chen et al. (2002). Site-directed mutagenesis was performed using Gene Tailor site-directed mutagenesis kit. The C93S Trx2 mutant was generated by hybridization of the plasmid with oligonucleotides of the sequences 5'-ACGCACAGTGGTGTGGACCCAGCAAGATCCTG-3' and 5'-GGGTCCACAC CACTGTGCGTGGAAATCCAC-3'. Mutations were confirmed by DNA sequencing analysis. Western blot analysis for changes in expression with transfection was performed previously and showed an increase of approximately 250% for both Trx2 and C93S Trx2 and 280% for Trx1 (data not shown).
NF-B activity.
HeLa cells were treated with TNF- (20 ng/ml) for 2 h. Nuclear translocation of NF-B was determined by collection of nuclear protein fractions using the NE-PER nuclear fractionation kit (Pierce Biotechnology, Rockford, IL). Nuclear proteins were separated on a 15% sodium dodecyl sulfate–polyacrylamide gel by electrophoresis and detected by immunoblotting. Both p50 and p65 NF-B subunits were immunodetected with rabbit anti-p50 and mouse anti-p65 primary antibodies (Santa Cruz Biotechnology, Santa Cruz, CA), respectively. Donkey anti-goat 800 (Rockland Immunochemicals, Gilberstville, PA) and goat anti-mouse AlexaFluor 680 antibodies (Invitrogen) were used for secondary detection.
HeLa cells were transfected with either empty, Trx2, or C93S Trx2 vectors and cotransfected with a NF-B-luciferase (NF-B-luc) reporter construct (Invitrogen). HeLa cells were treated for 6 h with TNF- (20 ng/ml) to accentuate luciferase expression differences. Lysates were collected in report lysis buffer (Promega, Madison, WI), followed by freeze-thaw, centrifugation, and storage of the supernatant at –70°C until assayed. Luciferase assays were performed as per the manufacturer's instructions using the luciferase assay system (Promega).
Terminal dUTP nick-end labeling assay.
Terminal dUTP nick-end labeling (TUNEL) was performed to determine TNF-–induced apoptosis. HeLa cells were transfected with Trx2 or C93S Trx2 prior to treatment. Cultures were treated with cyclohexamide (10 µg/ml) 1 h before the addition of TNF- (20 ng/ml) for 24 h. Prior treatment with cyclohexamide was necessary to inhibit the proapoptotic effect of TNF- and induced apoptosis. Cyclohexamide was omitted from other experiments so as to minimize variable-affected ROS generation, but as cyclohexamide has been primarily used to evaluate apoptosis, it was only used for the analysis of TNF-–induced apoptosis. Following treatment, cells were washed and fixed with 4% paraformaldehyde for 20 min. The TUNEL assay was performed using in situ cell death detection Kkt (Roche, Indianapolis, IN) as per the manufacturer's instructions. Cells were counterstained with 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride and were visualized by fluorescence microscopy. Random fields (five fields per experiment) were analyzed, and total cells were counted. Cells that showed punctate green fluorescence localized to the nucleus were designated as TUNEL-positive cells. Data are represented as percentage of TUNEL-positive cells of total cells.
Statistics.
Each measurement is the result of at least three independently performed experiments. The one-way analysis of variance was employed to determine whether the means of different groups were significantly different. The Tukey's post hoc test was used to determine the significance for all pairwise comparisons of interest.
|
RESULTS |
|---|
|
TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
Trx2, but Not Trx1, Redox State Is Oxidized by TNF-
Due to the independence of the Trx1 and Trx2 systems, measurement of oxidation of Trx1 and Trx2 allows for the determination of compartmental ROS generation and changes in redox environments. Cells were treated with increasing concentrations of TNF-
(0–40 ng/ml) for 30 min, to best differentiate early redox events and origin of ROS generation. Redox analysis of Trx1 showed that TNF- treatments produced no significant increase in oxidation (Fig. 1A), where Trx1 redox potential (Eh) were maintained between –270 and –255 mV. While very little effect was observed with TNF-–induced modulation of Trx1 Eh, Trx2 Eh was oxidized in a dose-dependent manner (Fig. 1B). The greatest oxidation as observed with 40 ng/ml TNF- was –322 mV, an increase in Eh of approximately 40 mV as compared to control.
|
The above experiments do not take into account transient changes in Trx1 and Trx2 redox states. TNF-
effects on both Trx1 and Trx2 were evaluated during the first 30 min of treatment and analyzed by redox Western techniques. TNF- time-course experiments showed that Trx1 was not oxidized within the first 30 min of TNF- treatment (Fig. 2A). However, Trx2 was significantly oxidized by TNF- treatment within 5 min, where the Eh increased by nearly 25 mV (Fig. 2B). The greatest oxidation of Trx2 by TNF- at the concentrations tested was observed at 30 min, where the Trx2 Eh increased by approximately 40 mV.
|
TNF-
Induces Mitochondrial ROS GenerationTNF-
caused a dose-dependent (0–40 ng/ml) increase in DCF fluorescence, suggesting an increase in total cellular ROS generation (Fig. 3A). These findings correlate with other findings showing TNF-–induced ROS generation (Goossens et al., 1995; Schulze-Osthoff et al., 1992). To determine mitochondrial ROS generation, we utilized the mitochondrial ROS detection reagent MitoSox Red. This approach would allow for the specific detection of mitochondrial ROS. TNF- treatments (0–40 ng/ml) caused an increase in MitoSox Red fluorescence, producing similar patterns as observed with DCF fluorescence.
|
Trx2 but Not Trx1 Reduces TNF-
–Induced ROS GenerationPrevious studies show that overexpression of either cytoplasmic Trx1 or mitochondrial Trx2 protects against oxidant-induced cell death (Andoh et al., 2002
; Chen et al., 2002). Thus, if there is a compartmental specificity in TNF-–induced ROS generation, this difference may be detectable by overexpressing Trx1 and Trx2 in cells loaded with ROS indicators. Results show that Trx2 overexpression blocked an increase in TNF-–induced DCF fluorescence, while Trx1 did not (Fig. 4). Confirmation of the Trx2-mediated ROS detoxification was demonstrated by overexpression of a dominant negative Trx2 (C93S Trx2). Overexpression of C93S Trx2 did not block an increase in ROS generation, suggesting that TNF- stimulation causes an increase in mitochondrial ROS formation and that Trx2 is primarily involved in this ROS detoxification and Trx1 is not.
|
Trx2 Inhibits TNF-
–Induced NF-B ActivationROS specifically from the mitochondria can activate NF-
B (Garcia-Ruiz et al., 1995). TNF- induction of NF-B results in the expression of several antiapoptotic proteins (Beg and Baltimore, 1996; Van Antwerp et al., 1996), which are believed to account for TNF-'s pathological effects (Aggarwal, 2003). NF-B readily translocates to the nucleus upon activation. As a measure of NF-B activation, nuclear isolates were measured for both the p50 and p65 NF-B subunits from cells that were transfected with empty, Trx2, or C93S Trx2 expression vectors. Cultures transfected with the empty vector and treated with TNF- showed an increase of both the p50 and p65 translocation to the nucleus, measured as an increase of 65 and 73%, respectively, as compared to nontreated samples (Fig. 5). Cultures transfected with the Trx2 expression vector and treated with TNF- showed no increase in translocation of either p50 or p65 as compared to empty vector controls treated with TNF-. C93S Trx2 transfection restored nuclear translocation of both the p50 and p65 subunits to similar levels as were seen in the TNF-–treated controls.
|
A reporter construct, NF-
B-luc, was used to determine NF-B activity. These results correlated with the previous findings showing that NF-B nuclear translocation was largely inhibited by Trx2 overexpression. NF-B activity was decreased by Trx2 as compared to controls by approximately 50% in cultures without TNF- stimulation (Fig. 6), but overexpression of C93S showed only a small increase. TNF- treatment in cultures transfected with an empty vector showed a substantial increase in NF-B activity as compared to nontransfected control (data not shown). Trx2 overexpression inhibited TNF-–induced NF-B activity by 50% as compared to TNF-–induced controls. ROS appears to be necessary for NF-B activation and gene regulation. Overexpression of Trx2 and the resulting decrease in ROS reduce NF-B activity and the subsequent gene regulation, suggesting that trx2 is an important factor in regulating mitochondria-dependent TNF- signaling.
|
Trx2 Inhibits TNF-
–Induced ApoptosisTo test the role of Trx2 in TNF-
–induced apoptosis, overexpression of Trx2 and the C93S dominant negative mutant was performed. Empty vector control cells that were not treated with TNF- showed low levels of apoptosis (0.4% [± 0.1] TUNEL positive) (Table 1). Cells transfected with the empty vector and treated with TNF- showed apoptosis measuring approximately 4.6% (± 0.6) cells being TUNEL positive. Cells transfected with Trx2 were more resistant to TNF-–induced apoptosis, where these cells showed significantly less apoptosis with TNF- treatment, measuring only 1.3% (± 0.4) cells being TUNEL positive. Cells transfected with C93S Trx2 had significantly elevated levels of apoptosis as compared to empty vector controls. Apoptosis in C93S Trx2 transfected cells measured 8.8% (± 0.9) TUNEL positive. These results show that mitochondrially derived ROS are necessary for TNF-–induced apoptosis.
|
|
DISCUSSION |
|---|
|
TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
TNF-
signaling is complex as it activates multiple pathways that are involved in both apoptosis and survival pathways. In apoptosis, TNF- promotes mitochondria swelling and changes in the mitochondrial permeability transition (MPT), which appear to be a consequence of TNF-–induced ROS generation (Bradham et al., 1998; Rutka et al., 1988; Zamzami et al., 1995). Converse to TNF- apoptotic pathways, TNF- activates NF-B, which regulates the expression of multiple genes that promote survival, including decoy receptor, BCL-XL, X-chromosome–linked inhibitor of apoptosis protein, FAS-associated death domainlike interleukin-1–converting enzyme inhibitory protein, TNFR-associated factor 1, and survivin (Karin and Lin, 2002). Beneficial effects of TNF- (i.e., tumor regression) appear to be largely countered by NF-B activation (Aggarwal, 2003). Thus, understanding factors that regulate both apoptosis and NF-B may contribute to future disease therapy.
Multiple studies show that TNF- is capable of altering the intracellular redox status (Ginn-Pease and Whisler, 1998; Schreck et al., 1991) and that TNF- may mediate its effects through ROS generation and the subsequent changes in redox potential (Ishii et al., 1992; Mehlen et al., 1996; Obrador et al., 1997; Phelps et al., 1995). In addition, the source of TNF-–induced ROS generation implicates the involvement of the mitochondria (Goossens et al., 1995; Shoji et al., 1995; Zamzami et al., 1995). Due to the importance of mitochondrially derived ROS, understanding mitochondrial antioxidant system function is critical as it is a primary regulator of TNF- downstream events.
Studies of mitochondrial antioxidant function have primarily focused on the role GSH, which may be very important as very little catalase activity is observed in the mitochondrial matrix (Reed, 1990). The importance of mitochondrial GSH is further supported by studies utilizing dimethyl maleate (DEM), which reacts with free sulfhydryls including GSH, depleting intracellular concentrations by over 90% (Goossens et al., 1995), and buthionine sulfoximine (BSO), which depletes GSH by inhibiting de novo synthesis. Following TNF- treatment and BSO pretreatment, ROS production did not change, but with TNF- treatment and DEM pretreatment, a 20-fold increase of ROS was observed (Goossens et al., 1995). BSO treatments have been shown to primarily deplete cytosolic GSH but not mitochondrial GSH (Griffith and Meister, 1985), while DEM appears to affect all GSH pools, including the mitochondrial GSH pool (Goossens et al., 1995). This evidence supports a role of GSH in regulating TNF-–induced ROS production, but these studies have largely ignored the importance of other mitochondrial redox couples, such as Trx2.
Although GSH is found in the mitochondria, to replenish mitochondrial pools, GSH must be synthesized in the cytosol and transported (Griffith and Meister, 1985). Inhibition of GSH transport from chronic ethanol exposure can exacerbate the effects of other oxidants, such as acetaminophen (Zhao and Slattery, 2002), and cytokines, such as TNF- (Brown et al., 2001). Trx's are a major redox system that is present in the mitochondria, nucleus, and cytosol. There are multiple functions of Trx1, including reduction of oxidized proteins (e.g., transcription factors), detoxification of ROS through Prx's, the synthesis of DNA, and cytokine function (Yodoi, 2000). The mitochondrial Trx2 system is independent of the cytosolic Trx1 system, supporting a mechanism independent of coping with cytosolic versus mitochondrial oxidative stress. The present results are consistent with Trx2 support elimination of ROS and thereby decreasing the amount of ROS available to act as a second messenger during TNF- stimulation. Thus, Trx2 overexpression decreased both NF-B activation and apoptosis. While these data are convincing, it does not discount the function or activity of other antioxidants, both inside and outside the mitochondria. Experiments were optimized for the study of Trx2 and its role in TNF- signaling. Early time points show that Trx2 is principally responsible for ROS detoxification, but it is possible that other antioxidant systems (i.e., Trx1 or GSH) could play an increasingly important role at later time points where mitochondrial ROS may permeate into the cytoplasm.
Trx1 has been shown to have oxidoreductase activities, reducing oxidized proteins and often restoring protein function. Although Trx2 has not been shown to perform this function, it is feasible as both Trx1 and Trx2 contain the same active-site motif (Cys-Gly-Pro-Cys) (Chen et al., 2002; Powis and Montfort, 2001). The adenine translocator is a critical component of the membrane permeability transition (MPT) pore and contains redox-sensitive cysteines that are susceptible to oxidation (Costantini et al., 2000; Vieira et al., 2001). Overexpression of Trx2 protects against oxidant-induced apoptosis, an effect that could be due to peroxide elimination or redox control of proteins involved in the MPT.
Unlike other antioxidant systems, such as Trx1 and GSH, Trx2 is specific to the mitochondria. Our data show that Trx2 manages TNF-–induced ROS generation in the mitochondria and as a result regulates downstream effects, such as NF-B activation and apoptosis. These data support the compartmentation of ROS signaling within the mitochondria but also suggest that other antioxidant systems, besides GSH, may play a role in the regulation of TNF-–mediated redox signaling.
|
ACKNOWLEDGMENTS |
|---|
This study was supported by National Institutes of Health grants R01 ES 011195 and F32 ES 013015.
|
REFERENCES |
|---|
|
TOPABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
|---|
Aggarwal, B. B. (2003). Signalling pathways of the TNF superfamily: A double-edged sword. Nat. Rev. Immunol. 3, 745–756.[CrossRef][ISI][Medline]
Andoh, T., Chock, P. B., and Chiueh, C. C. (2002). The roles of thioredoxin in protection against oxidative stress-induced apoptosis in SH-SY5Y cells. J. Biol. Chem. 277, 9655–9660.
Beg, A. A., and Baltimore, D. (1996). An essential role for NF-kappaB in preventing TNF-alpha-induced cell death. Science 274, 782–784.
Bellomo, G., Vairetti, M., Stivala, L., Mirabelli, F., Richelmi, P., and Orrenius, S. (1992). Demonstration of nuclear compartmentalization of glutathione in hepatocytes. Proc. Natl. Acad. Sci. U.S.A. 89, 4412–4416.
Bradham, C. A., Qian, T., Streetz, K., Trautwein, C., Brenner, D. A., and Lemasters, J. J. (1998). The mitochondrial permeability transition is required for tumor necrosis factor alpha-mediated apoptosis and cytochrome c release. Mol. Cell. Biol. 18, 6353–6364.
Brown, L. A., Harris, F. L., and Guidot, D. M. (2001). Chronic ethanol ingestion potentiates TNF-alpha-mediated oxidative stress and apoptosis in rat type II cells. Am. J. Physiol. Lung Cell. Mol. Physiol. 281, L377–L386.
Carswell, E. A., Old, L. J., Kassel, R. L., Green, S., Fiore, N., and Williamson, B. (1975). An endotoxin-induced serum factor that causes necrosis of tumors. Proc. Natl. Acad. Sci. U.S.A. 72, 3666–3670.
Cembrzynska-Nowak, M., Szklarz, E., Inglot, A. D., and Teodorczyk-Injeyan, J. A. (1993). Elevated release of tumor necrosis factor-alpha and interferon-gamma by bronchoalveolar leukocytes from patients with bronchial asthma. Am. Rev. Respir. Dis. 147, 291–295.[ISI][Medline]
Chen, Y., Cai, J., Murphy, T. J., and Jones, D. P. (2002). Overexpressed human mitochondrial thioredoxin confers resistance to oxidant-induced apoptosis in human osteosarcoma cells. J. Biol. Chem. 277, 33242–33248.
Costantini, P., Belzacq, A. S., Vieira, H. L., Larochette, N., de Pablo, M. A., Zamzami, N., Susin, S. A., Brenner, C., and Kroemer, G. (2000). Oxidation of a critical thiol residue of the adenine nucleotide translocator enforces Bcl-2-independent permeability transition pore opening and apoptosis. Oncogene 19, 307–314.[CrossRef][ISI][Medline]
Cotgreave, I. A. (2003). Analytical developments in the assay of intra- and extracellular GSH homeostasis: Specific protein S-glutathionylation, cellular GSH and mixed disulphide compartmentalisation and interstitial GSH redox balance. Biofactors 17, 269–277.[ISI][Medline]
Damdimopoulos, A. E., Miranda-Vizuete, A., Pelto-Huikko, M., Gustafsson, J. A., and Spyrou, G. (2002). Human mitochondrial thioredoxin. Involvement in mitochondrial membrane potential and cell death. J. Biol. Chem. 277, 33249–33257.
Garcia-Ruiz, C., Colell, A., Morales, A., Kaplowitz, N., and Fernandez-Checa, J. C. (1995). Role of oxidative stress generated from the mitochondrial electron transport chain and mitochondrial glutathione status in loss of mitochondrial function and activation of transcription factor nuclear factor-kappa B: Studies with isolated mitochondria and rat hepatocytes. Mol. Pharmacol. 48, 825–834.[Abstract]
Ginn-Pease, M. E., and Whisler, R. L. (1998). Redox signals and NF-kappaB activation in T cells. Free Radic. Biol. Med. 25, 346–361.[CrossRef][ISI][Medline]
Goossens, V., Grooten, J., De Vos, K., and Fiers, W. (1995). Direct evidence for tumor necrosis factor-induced mitochondrial reactive oxygen intermediates and their involvement in cytotoxicity. Proc. Natl. Acad. Sci. U.S.A. 92, 8115–8119.
Griffith, O. W., and Meister, A. (1985). Origin and turnover of mitochondrial glutathione. Proc. Natl. Acad. Sci. U.S.A. 82, 4668–4672.
Halvey, P. J., Watson, W. H., Hansen, J. M., Go, Y. M., Samali, A., and Jones, D. P. (2005). Compartmental oxidation of thiol-disulphide redox couples during epidermal growth factor signalling. Biochem. J. 386, 215–219.[CrossRef][ISI][Medline]
Hansen, J. M., Go, Y. M., and Jones, D. P. (2005). Nuclear and mitochondrial compartmentation of oxidative stress and redox signaling. Annu. Rev. Pharmacol. Toxicol. 46, 215–234.
Hansen, J. M., Watson, W. H., and Jones, D. P. (2004). Compartmentation of Nrf-2 redox control: Regulation of cytoplasmic activation by glutathione and DNA binding by thioredoxin-1. Toxicol. Sci. 82, 308–317.
Harper, R., Wu, K., Chang, M. M., Yoneda, K., Pan, R., Reddy, S. P., and Wu, R. (2001). Activation of nuclear factor-kappa b transcriptional activity in airway epithelial cells by thioredoxin but not by N-acetyl-cysteine and glutathione. Am. J. Respir. Cell Mol. Biol. 25, 178–185.
Higuchi, M., Proske, R. J., and Yeh, E. T. (1998). Inhibition of mitochondrial respiratory chain complex I by TNF results in cytochrome c release, membrane permeability transition, and apoptosis. Oncogene 17, 2515–2524.[CrossRef][ISI][Medline]
Hotamisligil, G. S., Peraldi, P., Budavari, A., Ellis, R., White, M. F., and Spiegelman, B. M. (1996). IRS-1-mediated inhibition of insulin receptor tyrosine kinase activity in TNF-alpha- and obesity-induced insulin resistance. Science 271, 665–668.[Abstract]
Ishii, Y., Partridge, C. A., Del Vecchio, P. J., and Malik, A. B. (1992). Tumor necrosis factor-alpha-mediated decrease in glutathione increases the sensitivity of pulmonary vascular endothelial cells to H2O2. J. Clin. Investig. 89, 794–802.[ISI][Medline]
Jones, D. P., Go, Y. M., Anderson, C. L., Ziegler, T. R., Kinkade, J. M., Jr, and Kirlin, W. G. (2004). Cysteine/cystine couple is a newly recognized node in the circuitry for biologic redox signaling and control. FASEB J. 18, 1246–1248.
Karin, M., and Lin, A. (2002). NF-kappaB at the crossroads of life and death. Nat. Immunol. 3, 221–227.[CrossRef][ISI][Medline]
Mehlen, P., Kretz-Remy, C., Preville, X., and Arrigo, A. P. (1996). Human hsp27, Drosophila hsp27 and human alphaB-crystallin expression-mediated increase in glutathione is essential for the protective activity of these proteins against TNFalpha-induced cell death. EMBO J. 15, 2695–2706.[ISI][Medline]
Merwin, J. R., Mustacich, D. J., Muller, E. G., Pearson, G. D., and Merrill, G. F. (2002). Reporter gene transactivation by human p53 is inhibited in thioredoxin reductase null yeast by a mechanism associated with thioredoxin oxidation and independent of changes in the redox state of glutathione. Carcinogenesis 23, 1609–1615.
Miyazaki, Y., Araki, K., Vesin, C., Garcia, I., Kapanci, Y., Whitsett, J. A., Piguet, P. F., and Vassalli, P. (1995). Expression of a tumor necrosis factor-alpha transgene in murine lung causes lymphocytic and fibrosing alveolitis. A mouse model of progressive pulmonary fibrosis. J. Clin. Investig. 96, 250–259.[ISI][Medline]
Nkabyo, Y. S., Ziegler, T. R., Gu, L. H., Watson, W. H., and Jones, D. P. (2002). Glutathione and thioredoxin redox during differentiation in human colon epithelial (Caco-2) cells. Am. J. Physiol. Gastrointest. Liver Physiol. 283, G1352–G1359.
Obrador, E., Navarro, J., Mompo, J., Asensi, M., Pellicer, J. A., and Estrela, J. M. (1997). Glutathione and the rate of cellular proliferation determine tumour cell sensitivity to tumour necrosis factor in vivo. Biochem. J. 325(Pt 1), 183–189.[Medline]
Phelps, D. T., Ferro, T. J., Higgins, P. J., Shankar, R., Parker, D. M., and Johnson, A. (1995). TNF-alpha induces peroxynitrite-mediated depletion of lung endothelial glutathione via protein kinase C. Am. J. Physiol. 269, L551–L559.[Medline]
Powis, G., and Montfort, W. R. (2001). Properties and biological activities of thioredoxins. Annu. Rev. Biophys. Biomol. Struct. 30, 421–455.[CrossRef][ISI][Medline]
Reed, D. J. (1990). Glutathione: Toxicological implications. Annu. Rev. Pharmacol. Toxicol. 30, 603–631.[CrossRef][ISI][Medline]
Rutka, J. T., Giblin, J. R., Berens, M. E., Bar-Shiva, E., Tokuda, K., McCulloch, J. R., Rosenblum, M. L., Eessalu, T. E., Aggarwal, B. B., and Bodell, W. J. (1988). The effects of human recombinant tumor necrosis factor on glioma-derived cell lines: Cellular proliferation, cytotoxicity, morphological and radioreceptor studies. Int. J. Cancer 41, 573–582.[ISI][Medline]
Schreck, R., Rieber, P., and Baeuerle, P. A. (1991). Reactive oxygen intermediates as apparently widely used messengers in the activation of the NF-kappa B transcription factor and HIV-1. EMBO J. 10, 2247–2258.[ISI][Medline]
Schulze-Osthoff, K., Bakker, A. C., Vanhaesebroeck, B., Beyaert, R., Jacob, W. A., and Fiers, W. (1992). Cytotoxic activity of tumor necrosis factor is mediated by early damage of mitochondrial functions. Evidence for the involvement of mitochondrial radical generation. J. Biol. Chem. 267, 5317–5323.
Shoji, Y., Uedono, Y., Ishikura, H., Takeyama, N., and Tanaka, T. (1995). DNA damage induced by tumour necrosis factor-alpha in L929 cells is mediated by mitochondrial oxygen radical formation. Immunology 84, 543–548.[ISI][Medline]
Smith, J. B., and Haynes, M. K. (2002). Rheumatoid arthritis—A molecular understanding. Ann. Intern. Med. 136, 908–922.
Suenaert, P., Bulteel, V., Lemmens, L., Noman, M., Geypens, B., Van Assche, G., Geboes, K., Ceuppens, J. L., and Rutgeerts, P. (2002). Anti-tumor necrosis factor treatment restores the gut barrier in Crohn's disease. Am. J. Gastroenterol. 97, 2000–2004.[CrossRef][ISI][Medline]
Sundaresan, M., Yu, Z. X., Ferrans, V. J., Irani, K., and Finkel, T. (1995). Requirement for generation of H2O2 for platelet-derived growth factor signal transduction. Science 270, 296–299.
Trotter, E. W., and Grant, C. M. (2003). Non-reciprocal regulation of the redox state of the glutathione-glutaredoxin and thioredoxin systems. EMBO Rep. 4, 184–188.[CrossRef][ISI][Medline]
Van Antwerp, D. J., Martin, S. J., Kafri, T., Green, D. R., and Verma, I. M. (1996). Suppression of TNF-alpha-induced apoptosis by NF-kappaB. Science 274, 787–789.
Vieira, H. L., Belzacq, A. S., Haouzi, D., Bernassola, F., Cohen, I., Jacotot, E., Ferri, K. F., El Hamel, C., Bartle, L. M., Melino, G., et al. (2001). The adenine nucleotide translocator: A target of nitric oxide, peroxynitrite, and 4-hydroxynonenal. Oncogene 20, 4305–4316.[CrossRef][ISI][Medline]
Voehringer, D. W., McConkey, D. J., McDonnell, T. J., Brisbay, S., and Meyn, R. E. (1998). Bcl-2 expression causes redistribution of glutathione to the nucleus. Proc. Natl. Acad. Sci. U.S.A. 95, 2956–2960.
Wang, H., and Joseph, J. A. (1999). Quantifying cellular oxidative stress by dichlorofluorescein assay using microplate reader. Free Radic. Biol. Med. 27, 612–616.[CrossRef][ISI][Medline]
Watson, W. H., Chen, Y., and Jones, D. P. (2003a). Redox state of glutathione and thioredoxin in differentiation and apoptosis. Biofactors 17, 307–314.[ISI][Medline]
Watson, W. H., and Jones, D. P. (2003). Oxidation of nuclear thioredoxin during oxidative stress. FEBS Lett. 543, 144–147.[CrossRef][ISI][Medline]
Watson, W. H., Pohl, J., Montfort, W. R., Stuchlik, O., Reed, M. S., Powis, G., and Jones, D. P. (2003b). Redox potential of human thioredoxin 1 and identification of a second dithiol/disulfide motif. J. Biol. Chem. 278, 33408–33415.
Yodoi, J. (2000). Evolution of thioredoxin and redox signaling research: Viewpoint. Antioxid. Redox Signal. 2, 629–630.[Medline]
Zamzami, N., Marchetti, P., Castedo, M., Decaudin, D., Macho, A., Hirsch, T., Susin, S. A., Petit, P. X., Mignotte, B., and Kroemer, G. (1995). Sequential reduction of mitochondrial transmembrane potential and generation of reactive oxygen species in early programmed cell death. J. Exp. Med. 182, 367–377.
Zhao, P., and Slattery, J. T. (2002). Effects of ethanol dose and ethanol withdrawal on rat liver mitochondrial glutathione: Implication of potentiated acetaminophen toxicity in alcoholics. Drug Metab. Dispos. 30, 1413–1417.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  D. P. Jones Radical-free biology of oxidative stress Am J Physiol Cell Physiol, October 1, 2008; 295(4): C849 - C868. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. He, J. Cai, Y.-M. Go, J. M. Johnson, W. D. Martin, J. M. Hansen, and D. P. Jones Identification of Thioredoxin-2 as a Regulator of the Mitochondrial Permeability Transition Toxicol. Sci., September 1, 2008; 105(1): 44 - 50. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. E. Nowak, B. Tian, M. Jamaluddin, I. Boldogh, L. A. Vergara, S. Choudhary, and A. R. Brasier RelA Ser276 Phosphorylation Is Required for Activation of a Subset of NF-{kappa}B-Dependent Genes by Recruiting Cyclin-Dependent Kinase 9/Cyclin T1 Complexes Mol. Cell. Biol., June 1, 2008; 28(11): 3623 - 3638. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. B. Chiribau, L. Cheng, I. C. Cucoranu, Y.-S. Yu, R. E. Clempus, and D. Sorescu FOXO3A Regulates Peroxiredoxin III Expression in Human Cardiac Fibroblasts J. Biol. Chem., March 28, 2008; 283(13): 8211 - 8217. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. L. K. Lim, J. Liu, M. L. Go, and U. A. Boelsterli The Mitochondrial Superoxide/Thioredoxin-2/Ask1 Signaling Pathway is Critically Involved in Troglitazone-Induced Cell Injury to Human Hepatocytes Toxicol. Sci., February 1, 2008; 101(2): 341 - 349. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. Patwari and R. T. Lee Thioredoxins, Mitochondria, and Hypertension Am. J. Pathol., March 1, 2007; 170(3): 805 - 808. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Ramachandiran, J. M. Hansen, D. P. Jones, J. R. Richardson, and G. W. Miller Divergent Mechanisms of Paraquat, MPP+, and Rotenone Toxicity: Oxidation of Thioredoxin and Caspase-3 Activation Toxicol. Sci., January 1, 2007; 95(1): 163 - 171. [Abstract] [Full Text] [PDF]
|
|
| ||||||