XAV-939

Myocardial ischemia reperfusion injury (MIRI) remains a major clinical challenge. Polyphenols derived from Potentilla anserina L (PPA) were found to possess anti-hypoxic-ischemic in vitro. The aim of this study was to examine (1) whether this plant exerts a protective effect and (2) whether the underlying mechanisms involving inflammatory responses and the Wnt/β-catenin signaling pathway were involved in rats. Rats were randomly divided as follows (1): Sham operation (2); ischemia-reperfusion (MIRI) (3); polyphenols (PPA) (4); Wnt inhibitor group (XAV939). In cardiomyocytes (1), HE staining was used to examine morphology (2), enzyme-linked immunosorbent assay to determine interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) levels and (3) Western blot to measure protein expression of Wnt and β-catenin. HE staining showed in MIRI edema of cardiomyocytes, localized inflammatory cell infiltration, unclear outline of cells and cell fragmentation, and some cardiomyocytes were seen to have disordered arrangement of tissues. PPA markedly lowered the morphological alterations in MIRI. The levels of levels of IL-1β and TNF-α in MIRI were significantly elevated; however, PPA reduced these cytokine concentrations compared to MIRI. Western blot analysis demonstrated that MIRI increased the relative protein expression levels of Wnt and β-catenin. Treatment of MIRI with PPA resulted in a significant decrease in protein expression levels of Wnt and β-catenin. The involvement of the Wnt/β-catenin pathway in MIRI was further affirmed with the use Wnt inhibitor group (XAV939) that also diminished protein expression levels of Wnt and β-catenin.

Myocardial ischemia reperfusion (I/R) injury is the main pathological manifestation of coronary artery disease closely linked with adverse cardiovascular outcomes. Aquaporin 1 (AQP1) is a water molecule that has been reported to be highly expressed during the process of myocardial I/R injury. The aim of this research was to explore the role of AQP1 in myocardial I/R injury and the relevant mechanism of action. RT-qPCR and western blotting were used to detect AQP1 expression. CCK-8 method was used to detect cell viability. JC-1 dye, MitoSox-Red staining and ATP-Red 1 probe were respectively used to detect mitochondrial membrane potential, mitochondrial ROS (mtROS) and ATP synthesis. C11-BODIPY 581/591 probe and FerroOrange probe were respectively used to measure lipid reactive oxygen species (ROS) and Fe(²⁺). Seahorse XFe96 Analyser was used to detect oxygen consumption rate (OCR). Assay kits were used to estimate mitochondrial permeability transition pore (mPTP) opening, total iron and lipid peroxidation levels. Western blotting was used to detect the expression of ferroptosis, energy metabolism and Wnt/β-catenin pathway-related proteins. AQP1 expression was elevated in hypoxia/reoxygenation (H/R)-exposed H9c2 cells. Deficient AQP1 promoted the viability, ameliorated mitochondrial dysfunction, ferroptosis and energy metabolism disorder in H/R-injured H9c2 cells. Further, AQP1 deletion might activate Wnt/β-catenin pathway and XAV939, an inhibitor of Wnt signaling pathway could partially revert the influences of AQP1 knockdown on the viability, mitochondrial function, ferroptosis and energy metabolism in H/R-treated H9c2 cells. To be concluded, AQP1 interference might protect against H/R-induced mitochondrial dysfunction, ferroptosis and energy metabolism disorder in H9c2 cells via modulating Wnt/β-catenin pathway.