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Trichostatin A attenuates oxidative stress-mediated myocardial injury through the FoxO3a signaling pathway
Yunhui Guo1, Zhiping Li1, Canxia Shi1
1Department of Pharmacology, College of Basic Medical Sciences, Jilin University, Changchun, Jilin 130021, P.R. China.
Abstract:
Trichostatin A (TSA), a histone deacetylase inhibitor, is widely used as an anticancer drug. Recently, TSA has been shown to exert a protective effect on ischemia/reperfusion (I/R) injury; however, the underlying mechanisms remain unclear. Forkhead box O3a (FoxO3a), a unique FoxO family member, has been shown to attenuate myocardial injury by increasing resistance to oxidative stress in mice. The present study aimed to investigate whether TSA exerts its cardioprotective effects through the FoxO3a signaling pathway. For this purpose, healthy male Wistar rats were pre-treated with TSA for 5 days before they were subjected to ligation/relaxation of the left anterior descending branch of the coronary artery and to 30 min of ischemia, followed by 24 h of reperfusion. The activities of creatine kinase (CK), lactate dehydrogenase (LDH), aspartate aminotransferase (AST) and superoxide diamutase (SOD), as well as the malondialdehyde (MDA) levels were examined. The H9c2 rat myocardial cell line was cultured in 10% FBS-containing DMEM for 24 h. The cells were incubated with/without TSA (50 nmol/l) for 1 h and then incubated with/without H2O2 (400 µM) for 2 h. Reactive oxygen species (ROS) and mitochondrial membrane potential (Δψm) were measured by probe staining in the H9c2 cells. The expression of FoxO3a, mitochondrial SOD2 and catalase was quantified by western blot analysis. The levels of H3 and H4 acetylation of the FoxO3a promoter region were examined by chromatin immunoprecipitation assay. TSA significantly reduced the myocardial infarct size and the activities of serum LDH, AST and CK in the rats. TSA also decreased the levels of MDA and increased the activities of SOD in the myocardial tissue of the rats. Consistent with the reduced injury to the TSA-treated rats, TSA significantly reduced the H2O2-induced levels of ROS and increased Δψm. In addition, TSA increased the expression of FoxO3a, SOD2 and catalase, which may be related to increasing the level of H4 acetylation of the FoxO3a promoter region. Our results thus revealed that TSA protected the myocardium from oxidative stress-mediated damage by increasing H4 acetylation of the FoxO3a promoter region, and the expression of FoxO3a, SOD2 and catalase.
Insights
Trichostatin A (TSA) protects the heart from ischemia/reperfusion injury by activating the Forkhead box O3a (FoxO3a) pathway. This involves increasing H4 acetylation, which boosts the expression of antioxidant enzymes like SOD2 and catalase, reducing oxidative stress.
Area of Science:
- Cardiovascular Research
- Molecular Biology
- Pharmacology
Background:
- Histone deacetylase inhibitor Trichostatin A (TSA) shows anticancer properties and recent evidence suggests cardioprotective effects against ischemia/reperfusion (I/R) injury.
- The precise mechanisms underlying TSA's protective role in I/R injury are not fully understood.
- Forkhead box O3a (FoxO3a) is a key transcription factor involved in cellular defense against oxidative stress and has been shown to attenuate myocardial injury.
Purpose of the Study:
- To investigate whether TSA exerts its cardioprotective effects against I/R injury via the FoxO3a signaling pathway.
- To elucidate the molecular mechanisms by which TSA protects the myocardium from oxidative stress.
Main Methods:
- In vivo studies involved pre-treating Wistar rats with TSA before inducing myocardial I/R injury, followed by assessment of cardiac injury markers (CK, LDH, AST) and oxidative stress indicators (SOD, MDA).
- In vitro studies utilized H9c2 rat myocardial cells treated with TSA and hydrogen peroxide (H2O2) to measure reactive oxygen species (ROS) and mitochondrial membrane potential (Δψm).
- Molecular analyses included Western blot to quantify protein expression (FoxO3a, SOD2, catalase) and chromatin immunoprecipitation (ChIP) assay to assess H3 and H4 acetylation of the FoxO3a promoter region.
Main Results:
- TSA treatment significantly reduced myocardial infarct size and serum levels of cardiac injury markers (LDH, AST, CK) in rats.
- TSA decreased malondialdehyde (MDA) levels and increased superoxide dismutase (SOD) activity in myocardial tissue, indicating reduced oxidative stress.
- In H9c2 cells, TSA attenuated H2O2-induced ROS production and preserved mitochondrial membrane potential, while increasing the expression of FoxO3a, SOD2, and catalase, associated with enhanced H4 acetylation of the FoxO3a promoter.
Conclusions:
- TSA demonstrates significant cardioprotective effects against ischemia/reperfusion injury in both in vivo and in vitro models.
- These protective effects are mediated through the activation of the FoxO3a signaling pathway, leading to increased expression of antioxidant enzymes.
- TSA enhances H4 acetylation of the FoxO3a promoter region, thereby upregulating FoxO3a, SOD2, and catalase, which collectively combat oxidative stress-induced myocardial damage.
