Sulforaphane inhibits mitochondrial permeability transition and oxidative stress

doi:10.1016/j.freeradbiomed.2011.09.017

Free Radical Biology and Medicine

Volume 51, Issue 12, 15 December 2011, Pages 2164-2171

https://doi.org/10.1016/j.freeradbiomed.2011.09.017 Get rights and content

Abstract

Exposure of mitochondria to oxidative stress and elevated Ca²⁺ promotes opening of the mitochondrial permeability transition pore (PTP), resulting in membrane depolarization, uncoupling of oxidative phosphorylation, and potentially cell death. This study tested the hypothesis that treatment of rats with sulforaphane (SFP), an activator of the Nrf2 pathway of antioxidant gene expression, increases the resistance of liver mitochondria to redox-regulated PTP opening and elevates mitochondrial levels of antioxidants. Rats were injected with SFP or drug vehicle and liver mitochondria were isolated 40 h later. Respiring mitochondria actively accumulated added Ca²⁺, which was then released through PTP opening induced by agents that either cause an oxidized shift in the mitochondrial redox state or directly oxidize protein thiol groups. SFP treatment of rats inhibited the rate of pro-oxidant-induced mitochondrial Ca²⁺ release and increased expression of the glutathione peroxidase/reductase system, thioredoxin, and malic enzyme. These results are the first to demonstrate that SFP treatment of animals increases liver mitochondrial antioxidant defenses and inhibits redox-sensitive PTP opening. This novel form of preconditioning could protect against a variety of pathologies that include oxidative stress and mitochondrial dysfunction in their etiologies.

Highlights

► Liver mitochondria from sulforaphane injected rats are resistant to PTP opening. ► Peroxidase activity is elevated in liver mitochondria from sulforaphane injected rats. ► Mitochondrial malic enzyme is elevated in sulforaphane injected rats. ► Sulforaphane injection elevates mitochondrial glutathione and antioxidant enzymes. ► Nrf2 activated gene expression increases mitochondrial resistance to oxidative stress.

Section snippets

Chemicals and reagents

R,S-sulforaphane was purchased from LKT Laboratories (St. Paul, MN, USA). Mannitol, sucrose, EGTA, Hepes, Tris, potassium phosphate dibasic (K₂HPO₄), magnesium chloride (MgCl₂), malate, glutamate, oligomycin, carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone, adenosine 5′-diphosphate (ADP), bovine serum albumin (BSA), succinate, rotenone, calcium, dimethyl sulfoxide (DMSO), tert-butylhydroperoxide (tBOOH), phenylarsine oxide, horseradish peroxidase, and oxaloacetate were obtained from

Evidence for Nrf2 activation in livers from rats treated with sulforaphane

NAD(P)H quinone oxidoreductase 1 is an antioxidant enzyme whose gene and protein expression is generally up-regulated by compounds that activate the Nrf2/ARE pathway, including sulforaphane [38]. To verify that this Nrf2 pathway biomarker protein was elevated in rats 40 h after intraperitoneal injection of sulforaphane at 10 mg/kg, liver homogenates were used for immunoblot analysis. This dose of sulforaphane and the time between injection of drug and liver acquisition were based on what were

Discussion

This is the first in-depth study of the regulation of mitochondrial PTP opening and antioxidant-related mitochondrial proteins by a pharmacologic activator of the Nrf2 pathway of cytoprotective gene expression. The primary conclusion reached from this study is that treatment of rats with sulforaphane results in a robust inhibition of PTP opening in rat liver mitochondria triggered by several forms of oxidative stress. These forms include a shift in redox state induced through either

Acknowledgment

This study was supported by NIH RO1NS34152.

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Original ContributionSulforaphane inhibits mitochondrial permeability transition and oxidative stress

Abstract

Highlights

Section snippets

Chemicals and reagents

Evidence for Nrf2 activation in livers from rats treated with sulforaphane

Discussion

Acknowledgment

Biochem. Biophys. Res. Commun.

FEBS Lett.

J. Biol. Chem.

J. Mol. Cell. Cardiol.

Exp. Neurol.

Arch. Biochem. Biophys.

Toxicol. In Vitro

Hepatology

Biochimie

Hepatology

Biochem. Biophys. Res. Commun.

Free Radic. Biol. Med.

J. Biol. Chem.

Biochim. Biophys. Acta

Free Radic. Biol. Med.

Biochem. Biophys. Res. Commun.

Arch. Biochem. Biophys.

Arch. Biochem. Biophys.

Lipoperoxide plasma levels in patients with liver cirrhosis

Hepatogastroenterology

Time course and mechanism of oxidative stress and tissue damage in rat liver subjected to in vivo ischemia–reperfusion

J. Clin. Invest.

A pore way to die: the role of mitochondria in reperfusion injury and cardioprotection

Biochem. Soc. Trans.

Effect of acute ethanol exposure on cultured fetal rat hepatocytes: relation to mitochondrial function

Alcohol. Clin. Exp. Res.

Significance of plasma glutathione determination in patients with alcoholic and non-alcoholic liver disease

J. Gastroenterol. Hepatol.

Oxidative damage to mitochondria is mediated by the Ca2+-dependent inner-membrane permeability transition

Biochem. J.

Original Contribution
Sulforaphane inhibits mitochondrial permeability transition and oxidative stress

Oxidative damage to mitochondria is mediated by the Ca²⁺-dependent inner-membrane permeability transition