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Updated: Dec 9, 2025
In vitro Assessment of Myocardial Protection following Hypothermia-Preconditioning in a Human Cardiac Myocytes Model
Published on: October 27, 2020
RGFP966 inactivation of the YAP pathway attenuates cardiac dysfunction induced by prolonged hypothermic preservation
Xiao-He Zheng1, Lin-Lin Wang1,2, Ming-Zhi Zheng3
1Department of Basic Medicine Sciences, Zhejiang University School of Medicine, Hangzhou 310058, China.
Insights
The histone deacetylase 3 (HDAC3) inhibitor RGFP966 protects donor hearts during prolonged hypothermic preservation by reducing oxidative stress and apoptosis. This mechanism involves the YAP pathway, improving cardiac function post-preservation.
Area of Science:
- Cardiovascular Research
- Organ Preservation
- Molecular Biology
Background:
- Oxidative stress and apoptosis limit hypothermic preservation of donor hearts to 4-6 hours.
- Prolonged preservation is crucial for expanding the donor pool and improving transplant outcomes.
Purpose of the Study:
- To investigate the protective effects of the histone deacetylase 3 (HDAC3) inhibitor RGFP966 against cardiac injury during prolonged hypothermic preservation.
- To elucidate the underlying molecular mechanisms involving oxidative stress, apoptosis, and the YAP pathway.
Main Methods:
- Rat hearts underwent 12-hour hypothermic preservation in Celsior solution with or without RGFP966, followed by 60 minutes of reperfusion.
- Evaluated hemodynamic parameters, protein expression (Mst1, YAP), apoptosis (TUNEL), and oxidative stress markers (LDH, MDA, antioxidant enzymes).
- Utilized Verteporfin (VP), a YAP-TEAD inhibitor, to investigate the role of the YAP pathway.
Main Results:
- RGFP966 significantly improved cardiac function and inhibited hypothermic preservation-induced cardiac dysfunction.
- RGFP966 modulated Mst1 and YAP phosphorylation, increased nuclear YAP levels, and attenuated oxidative stress and apoptosis.
- The protective effects of RGFP966 were partially abolished by VP, indicating the involvement of the YAP pathway.
Conclusions:
- Supplementation with RGFP966 effectively attenuated cardiac dysfunction following prolonged hypothermic preservation.
- The protective mechanism involves the inhibition of oxidative stress and apoptosis, potentially mediated by the inactivation of the YAP pathway.
Abstract:
Oxidative stress and apoptosis are the key factors that limit the hypothermic preservation time of donor hearts to within 4-6 h. The aim of this study was to investigate whether the histone deacetylase 3 (HDAC3) inhibitor RGFP966 could protect against cardiac injury induced by prolonged hypothermic preservation. Rat hearts were hypothermically preserved in Celsior solution with or without RGFP966 for 12 h followed by 60 min of reperfusion. Hemodynamic parameters during reperfusion were evaluated. The expression and phosphorylation levels of mammalian STE20-like kinase-1 (Mst1) and Yes-associated protein (YAP) were determined by western blotting. Cell apoptosis was measured by the terminal deoxynucleotidyl-transferase (TdT)-mediated dUTP nick-end labeling (TUNEL) method. Addition of RGFP966 in Celsior solution significantly inhibited cardiac dysfunction induced by hypothermic preservation. RGFP966 inhibited the hypothermic preservation-induced increase of the phosphorylated (p)-Mst1/Mst1 and p-YAP/YAP ratios, prevented a reduction in total YAP protein expression, and increased the nuclear YAP protein level. Verteporfin (VP), a small molecular inhibitor of YAP-transcriptional enhanced associate domain (TEAD) interaction, partially abolished the protective effect of RGFP966 on cardiac function, and reduced lactate dehydrogenase activity and malondialdehyde content. RGFP966 increased superoxide dismutase, catalase, and glutathione peroxidase gene and protein expression, which was abolished by VP. RGFP966 inhibited hypothermic preservation-induced overexpression of B-cell lymphoma protein 2 (Bcl-2)-associated X (Bax) and cleaved caspase-3, increased Bcl-2 mRNA and protein expression, and reduced cardiomyocyte apoptosis. The antioxidant and anti-apoptotic effects of RGFP966 were cancelled by VP. The results suggest that supplementation of Celsior solution with RGFP966 attenuated prolonged hypothermic preservation-induced cardiac dysfunction. The mechanism may involve inhibition of oxidative stress and apoptosis via inactivation of the YAP pathway.
