Improved Myocardial Function With Supplement Of Resveratrol To Celsior Solution

BackgroundHeart transplantation is the preferred treatment for the patients with severe heart failure and coronary artery disease. Successful organ preservation is more imperative to reduce the reperfusion injury, and is prerequisite for successful heart transplantation in clinical heart transplantation practice. Although many researchers have been reported that the cardiac function recovery after hypothermia preservation can be improved through the adjustment of heart preservation solution components, for example by adding oxygen free radical scavenger or calcium antagonists. Actually, heart hypothermia preservation in vitro for clinical transplantation is limited to approximately 6 hours. So how to prolong the preservation period and reduce the heart preservation-induced injury via improving the preservation solution is a problem urgently needed to be resolved.Resveratrol (3,5,4'-trihydroxy-trans-stilbene), a non-flavonoid polyphenol, is first been exacted from the roots of Veratrum album L. by a Japanese scientist. Naturally, resveratrol is abundantly present in grapes and mulburrys. Many studies have demonstrated that it elicits a broad spectrum of biological responses in vitro and in vivo, including anticarcinogenic effects, prolongation of life span, neuroprotective and cardioprotective activities, due to its antioxidant and anti-inflammation properties. So we speculated that resveratrol might also have the cardioprotection to prolong the preservation time, improve the cardiac function recovery after heart hypothermia preservation, which could make it possible to obtain donor heart from goodish distance and expand the possible donor heart sources.ObjectivesIn the present study, we aimed to determine the effect of resveratrol on cardiac dysfunction in hypothermic preservation rat hearts, and to explore its underlying mechanism.Methods(1) Establishment of hypothermic preservation rat heart model and measurement of cardiac functionAfter anesthesia, the hearts from male Sprague-Dawley rat were mounted on a Langendorff apparatus for perfusion at 37? with Krebs-Henseleit buffer. After 30 minutes of perfusion for stabilization, the hearts were retrograde perfused with cold Celsior preservation solution from aortic root to induce cardiac arrest. Then the hearts were moved to the cold Celsior preservation solution with or without resveratrol (10,20, 40 ?M) for 9 h, which followed by 60 min of reperfusion. During the reperfusion, Hemodynamic parameters (left ventricular end-diastolic pressure, left ventricular developed pressure,+dP/dtmax,-dP/dtmax, heart rate, and coronary flow) were monitored.(2) Western blottingAfter the experiment, the left ventricular muscle of rat heart was cut and lysed in ice-cold lysis buffer, then centrifuged at 4?. The supernatant containing 20 ?g of protein was collected for 10%SDS-polyacrylamide gel electrophoresis, then blotted onto a nitrocellulose membrane. After blocking, the membrane was incubated with UCP2 primary antibody (1:1000) overnight at 4?. After thorough washing, the membrane was then incubated with the horseradish peroxidase-conjugated secondary antibody for 2 h at room temperature. An enhanced ECL kit was used to detect the target protein.Results(1) After hypothermic preservation for 9 h followed by 60 min of reperfusion, the LVEDP increased, the recovery of LVDP,+dP/dtmax and -dP/dtmax decreased, heart rate and coronary flow declined in SD rat heart. Celsior solution supplement with resveratrol (10,20,40?M) could inhibit the hypothermic preservation-induced LVEDP elevation, prevent the hypothermic preservation-induced decrease in LVDP,+dP/dtmax,-dP/dtmax, heart rate and coronary flow in a dose-dependent manner.(2) Compared with control group, the UCP2 protein increased (3.2 fold) in rat heart suffered with 9 h of hypothermic preservation. Celsior solution supplement with resveratrol (20,40?M) could inhibit the overexpression of UCP2 protein induced by hypothermic preservation.(3) Compared with Celsior group, a SIRT1 inhibitor EX-257 had no significantly effects on the UCP2 protein expression and cardiac function in hypotherimic preservation rat heart. However, EX-257 could prevent the resveratrol-induced inhibition of UCP2 overexpression, and abolish the resveratrol-induced protection against cardiac dysfunction in the hypothermic preservation rat heart.ConclusionThe data suggested that Celsior solution supplement with resveratrol improved cardiac function recovery in hypothermic preservation rat hearts. The cardioprotective effects of resveratrol were found to be dependent on its ability to activate SIRT1 protein and inhibit hypothermic preservation-induced UCP2 protein overexpression.