Membrane receptor‐dependent Notch1/Hes1 activation by melatonin protects against myocardial ischemia–reperfusion injury: in vivo and in vitro studies

Abstract

Melatonin confers profound protective effect against myocardial ischemia-reperfusion injury (MI/RI). Activation of Notch1/Hairy and enhancer of split 1 (Hes1) signaling also ameliorates MI/RI. We hypothesize that melatonin attenuates MI/RI-induced oxidative damage by activating Notch1/Hes1 signaling pathway with phosphatase and tensin homolog deleted on chromosome 10 (Pten)/Akt acting as the downstream signaling pathway in a melatonin membrane receptor-dependent manner. Male Sprague Dawley rats were treated with melatonin (10 mg/kg/day) for 4 wk and then subjected to MI/R surgery. Melatonin significantly improved cardiac function and decreased myocardial apoptosis and oxidative damage. Furthermore, in cultured H9C2 cardiomyocytes, melatonin (100 μmol/L) attenuated simulated ischemia-reperfusion (SIR)-induced myocardial apoptosis and oxidative damage. Both in vivo and in vitro study demonstrated that melatonin treatment increased Notch1, Notch1 intracellular domain (NICD), Hes1, Bcl-2 expressions, and p-Akt/Akt ratio and decreased Pten, Bax, and caspase-3 expressions. However, these protective effects conferred by melatonin were blocked by DAPT (the specific inhibitor of Notch1 signaling), luzindole (the antagonist of melatonin membrane receptors), Notch1 siRNA, or Hes1 siRNA administration. In summary, our study demonstrates that melatonin treatment protects against MI/RI by modulating Notch1/Hes1 signaling in a receptor-dependent manner and Pten/Akt signaling pathways are key downstream mediators.

Melatonin ameliorates myocardial ischemia/reperfusion injury in type 1 diabetic rats by preserving mitochondrial function: role of AMPK-PGC-1α-SIRT3 signaling

“…Previously, we and others demonstrated that melatonin membrane receptors mediated the cardiac protective actions of melatonin2833. Moreover, we recently demonstrated that melatonin reduced flow shear stress (FSS)-induced bone marrow mesenchymal stem cell injury by activating melatonin membrane receptors and AMPK signaling50.…”

“…As an endogenous circadian hormone, melatonin has been proved to be a promising antioxidant against MI/R injury due to its strong anti-oxidative capacity and high safety profile928293031. Previously, we and others also demonstrated that melatonin exerted a solid protective effect against MI/R injury in non-diabetic animals by activating multiple intracellular signaling pathways such as silent information regulator 1 (SIRT1), Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) and Notch1/Hairy and enhancer of split 1293233.…”

“…As previously described2829, H9c2 cells were incubated in high-glucose medium (33 mmol/l) for 6 hours before the SIR treatment and during the entire reperfusion period (4 hours) to mimic the in vivo diabetic setting. Normal medium (containing 5.5 mmol/l glucose) was used as a control.…”

Ubiquitin-Specific Protease 4 Is an Endogenous Negative Regulator of Pathological Cardiac Hypertrophy

“…All the studies involving human samples were approved by the Ethics Committee of Renji Hospital, School of Medicine, Shanghai Jiao tong University, and conformed to the principles outlined in the Declaration of Helsinki. An expanded Materials and Methods section is available in the online-only Data Supplement, which includes detailed information on the following aspects: generation of global USP4 knockout and USP4 transgenic mice, surgical procedure of AB, 21 histological analysis, echocardiographic measurement, 22 culture of neonatal rat ventricular myocytes and adenovirus transfection, 23,24 immunofluorescence analysis, 25,26 ubiquitination assay, 27 immunoprecipitation, Western blot and quantitative real-time polymerase chain reaction, and human heart samples.…”

Novel Protective Role for Ubiquitin-Specific Protease 18 in Pathological Cardiac Remodeling

“…NRCMs were assayed by immunofluorescence studies, 19 and we observed that the overexpression of USP18 inhibited cardiomyocyte hypertrophy, whereas silencing USP18 exaggerated the hypertrophic response ( Figure 2B through 2F). Meanwhile, USP18 overexpression inhibited atrial natriuretic factor and β-MHC expression, whereas USP18 deficiency augmented it ( Figure 2E and 2F).…”