Resveratrol protects cardiomyocytes from hypoxia-induced apoptosis through the SIRT1–FoxO1 pathway

doi:10.1016/j.bbrc.2008.11.110

Biochemical and Biophysical Research Communications

Volume 378, Issue 3, 16 January 2009, Pages 389-393

https://doi.org/10.1016/j.bbrc.2008.11.110 Get rights and content

Abstract

Loss of cardiomyocytes through apoptosis has been proposed as a cause of ventricular remodeling and heart failure. Ischemia- and hypoxia-induced apoptosis of cardiomyocytes reportedly plays an important role in many cardiac pathologies. We investigated whether resveratrol (Res) has direct cytoprotective effects against ischemia/hypoxia for cardiomyocytes. Exposure of H9c2 embryonic rat heart-derived cells to hypoxia for 24 h caused a significant increase in apoptosis, as evaluated by TUNEL and flow cytometry, while treatment with 20 μM Res greatly decreased hypoxia-induced apoptosis in these cells. Exposure of the cells to Res (20 μM) caused rapid activation of SIRT1, which had a dual effect on FoxO1 function: SIRT1 increased FoxO1’s ability to induce cell cycle arrest, but inhibited FoxO1’s ability to induce cell death. This effect could be reversed by SIRT1 inhibition. Results of our study indicate that Res inhibits hypoxia-induced apoptosis via the SIRT1–FoxO1 pathway in H9c2 cells. This polyphenol may have potential in preventing cardiovascular disease, especially in coronary artery disease (CAD) patients.

Section snippets

Materials and methods

H9c2 embryonic rat heart-derived cells were obtained from American Type Culture Collection. Resveratrol, nicotinamide, and propidium iodide (PI) were purchased from Sigma. Antibodies against SIRT1, FoxO1, and BIM were purchased from Santa Cruz Biotechnology. Antibody against P27^KIP1 was purchased from Sigma. Alexa Fluor 555 anti-rabbit secondary antibody was purchased from Invitrogen. The In Situ Cell Apoptosis Detection kit was purchased from Promega.

Cell culture and treatments. H9c2 cells

Resveratrol protected H9c2 cells from apoptosis

The apoptosis of H9c2 cells was detected by TUNEL assay. As shown in Fig. 1, the percentage of TUNEL-positive cells was significantly greater in the ones incubated during 24 h under hypoxic conditions than normal control group (Cont: 0.37 ± 0.12% vs. Hyp: 11.60 ± 1.41%; P < 0.05). However, incubation with resveratrol (20 μM) for 24 h prevented most of the increase in apoptosis caused by hypoxia (Res: 4.76 ± 0.63% vs. Hyp; P < 0.05). Similar results were obtained with PI staining (Fig. 2).

Resveratrol activated SIRT1 in H9c2 cells

To explore the

Discussion

Studies have shown that apoptosis occurs in cardiomyocytes exposed to ischemia, but the mechanism and signaling pathway have been poorly elucidated to date. The NAD⁺-dependent protein deacetylase Sir2 is important for many cellular processes including gene silencing, cell cycle regulation, and life span extension [21], [22]. SIRT1, the most extensively studied human homolog of Sir2, has been reported to play a cardioprotective role because increased SIRT1 expression can circumvent cell

Acknowledgment

This work was supported by Natural Science Foundation of Guangdong Province, China (5008363).

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¹: These authors contributed equally to this work.

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Resveratrol protects cardiomyocytes from hypoxia-induced apoptosis through the SIRT1–FoxO1 pathway

Abstract

Section snippets

Materials and methods

Resveratrol protected H9c2 cells from apoptosis

Resveratrol activated SIRT1 in H9c2 cells

Discussion

Acknowledgment

FEBS Lett.

Biochem. Biophys. Res. Commun.

Biochem. Biophys. Res. Commun.

Cell

Mol. Cell

Cell

Cell

Trends Cell Biol.

J. Biol. Chem.

J. Pharmacol. Sci.

Mutat. Res.

Cell

J. Biol. Chem.

J. Biol. Chem.

Mech. Ageing Dev.

J. Biol. Chem.