Pifithrin-alpha attenuates p53-mediated apoptosis and improves cardiac function in response to myocardial ischemia/reperfusion in aged rats.
ABSTRACT Ischemic cardiovascular disease is a common age-related disease. The p53-dependent cardiac myocyte apoptosis induced by myocardial ischemia/reperfusion (MI/R) is an important feature in the progression of ischemic heart disease. In the present studies, we hypothesized that inhibition of p53-dependent myocyte apoptosis may improve cardiac dysfunction in aged rats after MI/R. A dose (2.2 mg/kg, i.p.) of pifithrin-alpha (PFT), a p53 inhibitor, or saline was administered to 20-month-old male F344 rats, which were subjected to 30 min of myocardial ischemia by ligating the left main coronary artery, followed by release of the ligature and 4 h of reperfusion. Results of our experiments indicate that MI/R induced a significant decrease in cardiac output index (CI) and mean arterial blood pressure (MABP). Administration of PFT to aged rats 40 min before ischemia significantly improved CI and MABP during 3 to 4 h of reperfusion. The improvement of cardiac function was associated with a marked reduction in DNA fragmentation in the area at risk of the heart when compared with aged MI/R rats pretreated with saline. Interestingly, treatment with PFT 10 min after ischemia or 10 min after reperfusion had a similar protective effect on CI and MABP, but this effect did not reach statistical significance when compared with aged MI/R rats pretreated with saline. Treatment with PFT, however, did not influence plasma creatine kinase activity and the number of circulating leukocytes and infiltrated leukocytes in the area at risk of the heart. Moreover, results of Western blot show that pretreatment with PFT significantly attenuated the ratio of Bax to Bcl-2 in the area-at-risk tissue of the heart compared with that of rats pretreated with saline. Our results suggest that pretreatment with PFT significantly improved cardiac function. The mechanism of protective effect of PFT may involve the inhibition of p53 transcriptional function, thereby attenuating the p53/Bax-mediated myocyte apoptosis during the reperfusion period.
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ABSTRACT: While the tumor suppressor functions of p53 have long been recognized, the contribution of p53 to numerous other aspects of disease and normal life is only now being appreciated. This burgeoning range of responses to p53 is reflected by an increasing variety of mechanisms through which p53 can function, although the ability to activate transcription remains key to p53's modus operandi. Control of p53's transcriptional activity is crucial for determining which p53 response is activated, a decision we must understand if we are to exploit efficiently the next generation of drugs that selectively activate or inhibit p53.Cell 06/2009; 137(3):413-31. DOI:10.1016/j.cell.2009.04.037 · 33.12 Impact Factor
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ABSTRACT: Activation of the PI3K/AKT pathway protects the heart from ischaemia-reperfusion injury. Phosphatase and Tensin Homolog deleted on Chromosome10 (PTEN) is a negative regulator of this pathway. The hypothesis on which this thesis was based stated that inhibition of PTEN would confer protection against ischaemia-reperfusion injury. PTEN was reduced using: 1) a PTEN inhibitor, bpV(HOpic), 2) a mouse model of PTEN haploinsufficiency and 3) PTEN siRNA. The effects of PTEN reduction on ischaemia-reperfusion injury were investigated by using: 1) an isolated perfused heart model of ischaemia-reperfusion injury, 2) an isolated cardiomyocyte model of ROS induced mitochondria damage and 3) a cellular model of hypoxia-reoxygenation injury. No protection against ischaemia-reperfusion was observed in isolated perfused myocardium from C57BL/J6 mice, which were perfused with bpV(HOpic), or from PTEN+/-mice. Likewise, no protection against ROS induced mitochondrial damage was observed in isolated cardiomyocytes from the PTEN+/- mice. In these models an increase in AKT activity was recorded, however, this was not sufficient to confer cardioprotection. Similarly, H9c2 rat myoblast cells, silenced for PTEN expression using siRNA, were not protected against hypoxia-reoxygenation injury. Nevertheless, in isolated C57BL/J6 hearts perfused with bpV(HOpic) and in myocardium from PTEN+/- mice, when the PI3K/AKT pathway was stimulated by the cardioprotective intervention of ischaemic preconditioning a reduced threshold for protection was achieved. To conclude, the level of PTEN inhibition achieved in this study was not sufficient to bestow protection against simulated ischaemiareperfusion injury. However, it appears that reductions in PTEN can increase the sensitivity towards cardioprotection.
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ABSTRACT: Ischemic heart disease is a common age-related disease. Apoptotic cell death and inflammation are the major contributors to I/R injury. The mechanisms that trigger myocyte apoptosis and inflammation during myocardial I/R (MI/R) remain to be elucidated. Published data from our laboratory demonstrated that pretreatment of MI/R rats with pifithrin-alpha (PFT), a specific p53 inhibitor, reduced myocyte apoptosis and improved cardiac function compared with MI/R rats pretreated with saline at 4 h of reperfusion. In the present study, we investigated the effects of PFT on the occurrence of myocyte apoptosis and leukocyte transmigration in the later period of reperfusion. Aged (20-month-old) male F344 rats were subjected to 30 min of myocardial ischemia via ligature of the LCA, followed by 24 h of reperfusion. Pifithrin-alpha (2.2 mg/kg, intraperitoneally) or saline was administered to rats before ischemia. The results indicate that pretreatment of MI/R rats with PFT significantly decreased the percentage of infarct area to ischemic area (33 +/- 8 vs. 54 +/- 9, P < 0.05) and improved cardiac output (79 +/- 11 vs. 38 +/- 9 mL/min per 100 g body weight, P < 0.05) when compared with rats pretreated with saline at 24 h of reperfusion. The protective effects of PFT may involve the p53/Bax-mediated apoptosis because treatment of MI/R rats with PFT attenuated the ratio of Bax to Bcl2 (0.97 +/- 0.1 vs. 2.1 +/- 0.2, P < 0.05) and reduced myocyte apoptosis. Interestingly, inhibition of p53 transcriptional function by PFT alleviated leukocyte infiltration into the ischemic area of the heart (339 +/- 37 vs. 498 +/- 75 cells/10 high-power fields, P < 0.05). These data suggest that inhibition of p53 transcriptional function by PFT attenuates myocyte apoptosis and alleviates leukocyte transmigration at 24 h of reperfusion. The mechanisms by which p53 modulates leukocyte transmigration require further investigation.Shock (Augusta, Ga.) 03/2008; 30(5):545-51. DOI:10.1097/SHK.0b013e31816a192d · 2.73 Impact Factor