Korean Circ J.  2007 Sep;37(9):408-413. 10.4070/kcj.2007.37.9.408.

The Cardioprotective Effects of Resveratrol via Anti-Apoptosis in Hypoxic Injury of Myocardial Cells

Affiliations
  • 1Department of Pediatrics, School of Medicine, The Catholic University of Daegu, Daegu, Korea. grapfarm@hanmail.net
  • 2Department of Pediatrics, Dongguk University College of Medicine, Gyeongju, Korea.
  • 3Department of Opthalmology, Dongguk University College of Medicine, Gyeongju, Korea.

Abstract

BACKGROUND AND OBJECTIVES: Resveratrol (trans-3, 4', 5-trihydroxy-stilbene), a naturally occurring polyphenolic phytoalexin found abundantly in grape skins and red wines, has been reported to protect heart cells from ischemia/reperfusion (I/R) injury through its significant antioxidant properties. Apoptosis of cardiac myocytes is also involved in several cardiovascular diseases, but it remains unknown whether the protective effects of resveratrol in hypoxic myocardial cell injury are mediated via suppression of apoptosis. In this study, we investigated whether resveratrol confers cardioprotection against hypoxia via anti-apoptosis in a hypoxic model of cultured H9c2 cardiomyoblasts.
MATERIALS AND METHODS
H9c2 cardiomyoblasts were obtained from the Korean Cell Line Bank. The cultured cells were divided into four groups: a normal control group, a hypoxia group, a group treated with resveratrol (10 microgram/mL) before hypoxic insult, and a group treated with resveratrol (10 microgram/mL) after hypoxic insult. The control group was placed in 5% CO2 incubators, and the hypoxia and resveratrol-treated groups were placed in 1% O2 incubators. Apoptosis was assayed by cytological analysis with Western blotting and real-time PCR for Bcl-2, Bax, and caspase-3.
RESULTS
The expression of Bcl-2 was significantly decreased in the hypoxia group compared with the control group, and resveratrol treatment inhibited the hypoxia-induced decline of Bcl-2 in hypoxic myocardial cells. Conversely, the expressions of Bax and caspase-3 were significantly increased in the hypoxia group, while resveratrol inhibited the hypoxia-induced increase of Bax and caspase. In addition, hypoxia significantly increased the ratio of Bax/Bcl-2 expression, but it was significantly decreased in the resveratrol-treated group.
CONCLUSION
The present study demonstrates that the cardioprotective effects of resveratrol in hypoxic injury are mediated via the mechanisms of anti-apoptosis.

Keyword

Resveratrol; Apoptosis; Bax protein; Bcl-2; Caspase-3

MeSH Terms

Anoxia
Apoptosis
bcl-2-Associated X Protein
Blotting, Western
Cardiovascular Diseases
Caspase 3
Cell Line
Cells, Cultured
Heart
Incubators
Myocytes, Cardiac
Real-Time Polymerase Chain Reaction
Skin
Vitis
Wine
Caspase 3
bcl-2-Associated X Protein

Figure

  • Fig. 1 High magnification (×400) photomicrographs of cultured H9c2 cells. A: control group; nuclear membranes are distinct and the shape of the cytoplasm is regular. B: hypoxia group; the picture shows cellular swelling. Damaged cells show cell membrane disruption and necrotic cellular debris. Nuclear shape is indistinct, the cytoplasm shows numerous irregular vacuoles. C: resveratrol group treated before hypoxic exposure; the cellular shape is generally regular. The degree of cellular swelling and number of damaged cells are decreased compared to those of (B, D). Resveratrol group treated after hypoxic exposure: findings of cellular swelling and damaged cells are similar to those of (B).

  • Fig. 2 Western blotting of Bcl-2 (A), Bax (B), and caspase-3 (C) from cultured H9c2 cells. The ratio of Bax/Bcl-2 (D) expression is also shown. Resveratrol was administered at 10 µg/mL. Immunoblots from three independent experiments are shown. Densitometric analysis is also shown. Data are presented as the ratios of band intensities compared with the control group. Before: resveratrol group treated before hypoxic exposure, After: resveratrol group treated after hypoxic exposure. *: p<0.05 compared with control.

  • Fig. 3 Real-time PCR was performed for Bcl-2 (A), Bax (B), and caspase-3 (C) on DNA from cultured H9c2 cells. The ratio of Bax/Bcl-2 (D) expression is also shown. Resveratrol was administered at 10 µg/mL. Data are presented as the ratios of mRNA expression compared with those of the control group. Before: resveratrol group treated before hypoxic exposure, After: resveratrol group treated after hypoxic exposure. *: p<0.05 compared with control. PCR: polymerase chain reaction, DNA: deoxyribonucleic acid, mRNA: messenger ribonucleic acid.


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