Antiapoptotic effect of Vitis vinifera (grape seed) proanthocyanidins in cardiomyocytes

Shao Z^1,3, Yuan CS², Becker LB^1,3, Schumacker PT^1,4, Hsu C^1,5, Wojick K^1,3, Li J^1,3, Li C^1,3, Anderson T^1,3, Qin Y^1,4, Hamann K^1,4, Barth E⁶, Halpern H⁶, Vanden Hoek TL^1,3

¹Emergency Resuscitation Center, University of Chicago, Chicago, IL 60637, USA

²Tang Center for Herbal Medicine Research, University of Chicago, Chicago, IL 60637, USA

³Department of Medicine, Section of Emergency Medicine, University of Chicago, Chicago, IL 60637, USA

⁴Department of Medicine, Section of Pulmonary/Critical Care, University of Chicago, Chicago, IL 60637, USA

⁵Tri-service General Hospital, National Defense Medical Center, Taiwan

⁶Radiation and Oncology, Center for Low Frequency EPR Imaging for In Vivo Physiology, University of Chicago, Chicago, IL 60637, USA

Objective

Grape seed proanthocyanidin extract (GSPE) treatment improves cardiomyocyte survival following ischemia/reperfusion (I/R). Since I/R injury in this system appears to be mediated in part via caspase activation while nitric oxide (NO) can be cardioprotective, we hypothesised that GSPE may decrease caspase-mediated I/R injury in chick cardiomyocytes via increased NO generation.

Materials and methods

Cultured chick cardiomyocytes were perfused and monitored in a Sykes–Moore chamber during 1 h of simulated ischemia and 3 h of reperfusion. Oxidant generation was measured by 2′, 7′-dichlorifluorescin diacetate (DCFH/DA). NO production was determined by 4′,5′-diaminofluorescein diacetate (DAF-2DA). Viability was assessed by propidium iodide (PI). Caspase activity was measured by fluorogenic assay.

Results

GSPE (10, 50 and 100 μg/ml) administered at the reperfusion dose-dependently attenuated the burst of DCF fluorescence. A significant reduction in cell death was observed from 44.9 ± 2.3% in I/R untreated cells to 17.8 ± 3.0% in GSPE-treated cells (50 μg/ml; n = 6, P < 0.001), with the return of contractions observed in the GSPE group. GSPE (50 μg/ml) significantly inhibited caspase 3, 2, 8, 9 activities compared to I/R untreated cells. When GSPE (50 μg/ml) was added at reperfusion DAF-2 fluorescence significantly increased and N^G-nitro-L-arginine methyl ester (L-NAME), a NOS inhibitor, partially abolished caspase inhibition and abrogated GSPE-conferred protection against cell death (from 17.8 ± 3.0% to 33.5 ± 3.8%).

Conclusion

GSPE attenuates I/R-induced apoptosis in chick cardiomyocytes by inhibiting caspase activity. GSPE may mediate protection by stimulating NO production within the cardiomyocytes.

Acknowledgement

This work was supported by grants AT01575 and HL68951 from NIH, USA.

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