Resveratrol Attenuates Ischemia/Reperfusion Injury in Neonatal Cardiomyocytes and Its Underlying Mechanism

Abstract

This study was designed to investigate whether Resveratrol (Res) could be a prophylactic factor in the prevention of I/R injury and to shed light on its underlying mechanism. Primary culture of neonatal rat cardiomyocytes were randomly distributed into three groups: the normal group (cultured cardiomyocytes were in normal conditions), the I/R group (cultured cardiomyocytes were subjected to 2 h simulated ischemia followed by 4 h reperfusion), and the Res+I/R group (100 µmol/L Res was administered before cardiomyocytes were subjected to 2 h simulated ischemia followed by 4 h reperfusion). To test the extent of cardiomyocyte injury, several indices were detected including cell viability, LDH activity, Na+-K+-ATPase and Ca2+-ATPase activity. To test apoptotic cell death, caspase-3 activity and the expression of Bcl-2/Bax were detected. To explore the underlying mechanism, several inhibitors, intracellular calcium, SOD activity and MDA content were used to identify some key molecules involved. Res increased cell viability, Na+-K+-ATPase and Ca2+-ATPase activity, Bcl-2 expression, and SOD level. While LDH activity, capase-3 activity, Bax expression, intracellular calcium and MDA content were decreased by Res. And the effect of Res was blocked completely by either L-NAME (an eNOS inhibitor) or MB (a cGMP inhibitor), and partly by either DS (a PKC inhibitor) or Glybenclamide (a KATP inhibitor). Our results suggest that Res attenuates I/R injury in cardiomyocytes by preventing cell apoptosis, decreasing LDH release and increasing ATPase activity. NO, cGMP, PKC and KATP may play an important role in the protective role of Res. Moreover, Res enhances the capacity of anti-oxygen free radical and alleviates intracellular calcium overload in cardiomyocytes.

Resveratrol attenuates hypoxia/reoxygenation-induced Ca2+ overload by inhibiting the Wnt5a/Frizzled-2 pathway in rat H9c2 cells

“…Thus, the control of the intracellular Ca 2+ levels is a key factor in the treatment of reperfusion injury. A recent study demonstrated that resveratrol suppressed H 2 O 2 -induced Ca 2+ inflow and indicated that resveratrol attenuated H/R-induced cell injury and apoptosis by inhibiting the Ca 2+ overload (11). Consistent with the abovementioned study, Ca 2+ release was demonstrated to be significantly decreased when H/R cardiomyocytes were treated with resveratrol.…”

“…Studies have reported that resveratrol is able to attenuate ischemia-reperfusion injury, inhibit low-density lipoprotein oxidation and promote vasorelaxation, which clearly support its potential role for the prevention and therapy of MIRI (9,10). Additionally, Shen et al (11) identified that resveratrol alleviates the cardiomyocyte Ca 2+ overload induced by H 2 O 2 . However, the mechanisms underlying its effects against MIRI and Ca 2+ overload are not well established.…”

Preclinical and clinical evidence for the role of resveratrol in the treatment of cardiovascular diseases

“…As the myocardium is reperfused following an acute ischemic episode, the tissue is damaged, in part, by the generation of ROS [188]. Resveratrol has been shown to reduce lipid-peroxidation [163,189], reduce the levels of superoxide [46,190], and activate anti-oxidant defense mechanisms [161,189]. A recent proteomics study on Yorkshire miniswine implanted with an ameroid constrictor identified a number of differences in proteins expressed in the animals pre-treated with resveratrol [191].…”

Lacidipine attenuates TNF-α-induced cardiomyocyte apoptosis

“…Cardiomyocyte apoptosis is involved in various heart diseases, such as myocardial hypertrophy, heart failure and myocardial ischemia [6,7]. Apoptosis is positively and negatively regulated by the Bcl-2 protein family [7][8][9][10].…”