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Cited by 721Cardiac Ischemia and ReperfusionCell death mechanisms and regulationMitochondrial Function and Pathology

The DOI matches its Crossref record; 40 of the 40 references it cites were found in a registry or an index.

Reactive Oxygen Species Released from Mitochondria during Brief Hypoxia Induce Preconditioning in Cardiomyocytes

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Vanden Hoek, Terry L., Becker, Lance B., Shao, Zuohui, Li, Changqing, Schumacker, Paul T.·1998·Journal of Biological Chemistry
DOI 10.1074/jbc.273.29.18092
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Abstract

Reactive oxygen species (ROS) have been proposed to participate in the induction of cardiac preconditioning. However, their source and mechanism of induction are unclear. We tested whether brief hypoxia induces preconditioning by augmenting mitochondrial generation of ROS in chick cardiomyocytes. Cells were preconditioned with 10 min of hypoxia, followed by 1 h of simulated ischemia and 3 h of reperfusion. Preconditioning decreased cell death from 47 +/- 3% to 14 +/- 2%. Return of contraction was observed in 3/3 preconditioned versus 0/6 non-preconditioned experiments. During induction, ROS oxidation of the probe dichlorofluorescin (sensitive to H2O2) increased approximately 2.5-fold. As a substitute for hypoxia, the addition of H2O2 (15 micromol/liter) during normoxia also induced preconditioning-like protection. Conversely, the ROS signal during hypoxia was attenuated with the thiol reductant 2-mercaptopropionyl glycine, the cytosolic Cu,Zn-superoxide dismutase inhibitor diethyldithiocarbamic acid, and the anion channel inhibitor 4,4'-diisothiocyanato-stilbene-2,2'-disulfonate, all of which also abrogated protection. ROS generation during hypoxia was attenuated by myxothiazol, but not by diphenyleneiodonium or the nitric-oxide synthase inhibitor L-nitroarginine. We conclude that hypoxia increases mitochondrial superoxide generation which initiates preconditioning protection. Furthermore, mitochondrial anion channels and cytosolic dismutation to H2O2 may be important steps for oxidant induction of hypoxic preconditioning.

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Crossref · 10.1074/jbc.273.29.18092

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FIG. 1 Effect of hypoxic preconditioning on cell death from ischemia/reperfusion. Cardiomyocytes were allowed to equilibrate at base-line normoxic conditions for 40 min, then preconditioned with 10 min of hypoxia (PO 2 5 3 torr), followed by a 10-min return to normoxia. Non-preconditioned cells were equilibrated with normoxia at base line for 60 min. Compared with non-preconditioned cells, preconditioned cells had significantly less cell death following 1 h of ischemia and 3 h of reperfusion.
FIG. 1 Effect of hypoxic preconditioning on cell death from ischemia/reperfusion. Cardiomyocytes were allowed to equilibrate at base-line normoxic conditions for 40 min, then preconditioned with 10 min of hypoxia (PO 2 5 3 torr), followed by a 10-min return to normoxia. Non-preconditioned cells were equilibrated with normoxia at base line for 60 min. Compared with non-preconditioned cells, preconditioned cells had significantly less cell death following 1 h of ischemia and 3 h of reperfusion.CC-BY · From the PDF
FIG. 2 ROS generation during hypoxic preconditioning. Oxidation of DCFH to DCF was monitored in preconditioned versus nonpreconditioned cells. Hypoxia increased DCFH oxidation compared with normoxic control cells. DCFH oxidation occurred during hypoxic preconditioning, but not during the recovery period when O 2 was reintroduced. Thus, hypoxic preconditioning was associated with a transient increase in ROS generation.
FIG. 2 ROS generation during hypoxic preconditioning. Oxidation of DCFH to DCF was monitored in preconditioned versus nonpreconditioned cells. Hypoxia increased DCFH oxidation compared with normoxic control cells. DCFH oxidation occurred during hypoxic preconditioning, but not during the recovery period when O 2 was reintroduced. Thus, hypoxic preconditioning was associated with a transient increase in ROS generation.CC-BY · From the PDF
FIG. 3 Effect of N -nitro-L-arginine or 2-MPG on DCFH oxidation during hypoxic preconditioning. ROS generation during 10 min of hypoxic preconditioning, suggested by increased DCFH oxidation, was attenuated by 2-MPG (400 m M), added during base-line conditions for 40 min and hypoxic preconditioning. However, the NOS inhibitor Nnitro-L-arginine (100 m M) had the opposite effect, increasing DCFH oxidation and DCF fluorescence.
FIG. 3 Effect of N -nitro-L-arginine or 2-MPG on DCFH oxidation during hypoxic preconditioning. ROS generation during 10 min of hypoxic preconditioning, suggested by increased DCFH oxidation, was attenuated by 2-MPG (400 m M), added during base-line conditions for 40 min and hypoxic preconditioning. However, the NOS inhibitor Nnitro-L-arginine (100 m M) had the opposite effect, increasing DCFH oxidation and DCF fluorescence.CC-BY · From the PDF
FIG. 4 Effect of DPI or myxothiazol on DCFH oxidation during hypoxic preconditioning. DPI (10 m M) had no significant effect on DCFH oxidation during 10 min of hypoxic preconditioning. By contrast, myxothiazol produced a dose-dependent attenuation of DCFH oxidation during hypoxic preconditioning. Thus, the ROS generation of hypoxic preconditioning appears to originate from the mitochondrial electron transport chain.
FIG. 4 Effect of DPI or myxothiazol on DCFH oxidation during hypoxic preconditioning. DPI (10 m M) had no significant effect on DCFH oxidation during 10 min of hypoxic preconditioning. By contrast, myxothiazol produced a dose-dependent attenuation of DCFH oxidation during hypoxic preconditioning. Thus, the ROS generation of hypoxic preconditioning appears to originate from the mitochondrial electron transport chain.CC-BY · From the PDF
FIG. 5 Possible pathway of ROS generation and metabolism during hypoxic preconditioning. During hypoxic preconditioning, superoxide generated from the mitochondrial electron tranport chain may be degraded in the mitochondria or may reach the cytosol via anion channels. There, it may be degraded by Cu,Zn-SOD to H2 O2, which activates subsequent mediators of preconditioning leading to preconditioning protection ( GSH , reduced glutathione; GSSG , oxidized glutathione).
FIG. 5 Possible pathway of ROS generation and metabolism during hypoxic preconditioning. During hypoxic preconditioning, superoxide generated from the mitochondrial electron tranport chain may be degraded in the mitochondria or may reach the cytosol via anion channels. There, it may be degraded by Cu,Zn-SOD to H2 O2, which activates subsequent mediators of preconditioning leading to preconditioning protection ( GSH , reduced glutathione; GSSG , oxidized glutathione).CC-BY · From the PDF
FIG. 6 Effects of DDC on ROS generation during hypoxic preconditioning. DDC (1 mM) was added 20 min prior to and during 10 min hypoxic preconditioning. A , DCF fluorescence increases during hypoxic preconditioning were attenuated by DDC. B , ethidium fluorescence was increased by DDC. These results suggest that cytosolic SOD is involved in metabolizing superoxide generated by hypoxic preconditioning to H 2 O2.
FIG. 6 Effects of DDC on ROS generation during hypoxic preconditioning. DDC (1 mM) was added 20 min prior to and during 10 min hypoxic preconditioning. A , DCF fluorescence increases during hypoxic preconditioning were attenuated by DDC. B , ethidium fluorescence was increased by DDC. These results suggest that cytosolic SOD is involved in metabolizing superoxide generated by hypoxic preconditioning to H 2 O2.CC-BY · From the PDF
Image 7
CC-BY · From the PDF
FIG. 7 Effects of DDC given during hypoxic preconditioning on cell death from ischemia/reperfusion. The protection conferred by hypoxic preconditioning was abolished when DDC was added during base-line and hypoxic preconditioning stages. These results suggest that the metabolism of superoxide to H2O2 by cytosolic SOD is an important step in the pathway from ROS generation to preconditioning protection.
FIG. 7 Effects of DDC given during hypoxic preconditioning on cell death from ischemia/reperfusion. The protection conferred by hypoxic preconditioning was abolished when DDC was added during base-line and hypoxic preconditioning stages. These results suggest that the metabolism of superoxide to H2O2 by cytosolic SOD is an important step in the pathway from ROS generation to preconditioning protection.CC-BY · From the PDF
FIG. 8 Preconditioning with exogenous H2O > 2 and abolition of preconditioning with 2-MPG. Preconditioning-like protection was induced by exposure to H 2 O2 (15 m mol/liter) for 10 min during normoxia prior to ischemia/reperfusion. By contrast, the protection of hypoxic preconditioning was abolished when the thiol reductant 2-MPG (400 m M) was added during base-line and hypoxic preconditioning stages, resulting in the same amount of cell death after ischemia/reperfusion compared with non-preconditioned cells.
FIG. 8 Preconditioning with exogenous H2O > 2 and abolition of preconditioning with 2-MPG. Preconditioning-like protection was induced by exposure to H 2 O2 (15 m mol/liter) for 10 min during normoxia prior to ischemia/reperfusion. By contrast, the protection of hypoxic preconditioning was abolished when the thiol reductant 2-MPG (400 m M) was added during base-line and hypoxic preconditioning stages, resulting in the same amount of cell death after ischemia/reperfusion compared with non-preconditioned cells.CC-BY · From the PDF
FIG. 9 Effects of DIDS on ROS production during hypoxic preconditioning and on subsequent cell death during ischemia/ reperfusion. ROS generation and protection associated with hypoxic preconditioning was abolished when DIDS (200 m M) was added during 40 min of base-line conditions and during 10 min of hypoxic preconditioning. A , DCF fluorescence increases during hypoxic preconditioning were attenuated by DIDS. B , DIDS abolished the protection previously seen with hypoxic preconditioning. These results suggest that anion channels are an important component in the pathway from mitochondrial ROS generation to preconditioning protection.
FIG. 9 Effects of DIDS on ROS production during hypoxic preconditioning and on subsequent cell death during ischemia/ reperfusion. ROS generation and protection associated with hypoxic preconditioning was abolished when DIDS (200 m M) was added during 40 min of base-line conditions and during 10 min of hypoxic preconditioning. A , DCF fluorescence increases during hypoxic preconditioning were attenuated by DIDS. B , DIDS abolished the protection previously seen with hypoxic preconditioning. These results suggest that anion channels are an important component in the pathway from mitochondrial ROS generation to preconditioning protection.CC-BY · From the PDF

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Show all 40 references

Research works printed but not linked: 6

  1. Liang, 1996, Am. J. Physiol., 271, H1769
  2. Vanden Hoek, 1996, Am. J. Physiol., 270, H1334
  3. Bond, 1991, Res. Commun. Chem. Pathol. Pharm., 71, 195
  4. Murry, 1988, Circulation, 78, 77
  5. Pathak, 1995, Circulation, 92, I
  6. Larsson, 1989, Cancer Res., 49, 5627

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Metadata

Authors
Vanden Hoek, T. L., Becker, L. B., Shao, Z., Li, C., & Schumacker, P. T.
Affiliations
University of Chicago (US)
Corresponding author
Schumacker, Paul T.
ORCID
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
1998-07
Volume
273
Issue
29
Pages
18092-18098
ISSN
0021-9258 (ISSN-L)
DOI
10.1074/jbc.273.29.18092
OpenAlex
W2145047965
Cited by
721
References
40
Citations by year
2026: 5 · 2025: 5 · 2024: 7 · 2023: 4 · 2022: 4 · 2021: 15 · 2020: 10 · 2019: 8 · 2018: 11 · 2017: 8 · 2016: 29 · 2015: 16 · 2014: 31 · 2013: 33 · 2012: 26
FWCI
20.19
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of Biological Chemistry (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • National Institutes of Health (5k08hl003779-02, HL35440)
  • National Heart, Lung, and Blood Institute (HL 32646, HL-03779, R01HL035440, HL35440)
Topics
  • Health Sciences › Medicine › Cardiology and Cardiovascular Medicine › Cardiac Ischemia and Reperfusion
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cell death mechanisms and regulation
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Mitochondrial Function and Pathology
Keywords
reactive oxygen species, mitochondrial superoxide, hypoxia, cardioprotection, preconditioning, cardiomyocytes, ischemia-reperfusion injury, hydrogen peroxide, superoxide dismutase, DIDS, myxothiazol, nitric oxide synthase, cell death
MeSH terms
Animals, Cells, Cultured, Chick Embryo, Cytosol, Ditiocarb, Enzyme Inhibitors, Hydrogen Peroxide, Tiopronin, Methacrylates, Mitochondria, Heart, Myocardial Contraction, Onium Compounds, Superoxide Dismutase, Thiazoles, Cell Hypoxia, Cell Death, Reactive Oxygen Species, Nitric Oxide Synthase, Ischemic Preconditioning, Myocardial

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