Inhibition of myeloid differentiation factor 2 by MAC28 suppresses reactive oxygen species, inflammation and improves mitochondrial function, leading to improved cardiac function in prediabetic rats

Abstract Background Chronic inflammation involves in the left ventricular (LV) dysfunction in high-fat diet (HFD)-induced prediabetes, along with cardiac mitochondrial dysfunction. This involved an activation of myeloid differentiation factor 2 (MD2)/toll-like receptor 4 (TLR4) by lipopolysaccharide, leading to inflammatory cytokines production in the heart. MAC28 is a novel MD2 inhibitor, which had been shown to provide effects against LPS-induced cytokine secretion from macrophages. However, the potential benefits of MAC28 on the LV function and its underlying mechanisms in HFD-induced prediabetic rats are unknown. Purpose We tested the hypothesis that MAC28 improves LV function in prediabetic rats by reducing cardiac oxidative stress, inflammation, and cardiac mitochondrial dysfunction. Methods Male Wistar rats were fed either a normal diet (ND, n=8) or HFD (n=24) for 16 weeks. At week 12, HFD-fed rats developed prediabetes and LV dysfunction. At this time, these HFD-fed rats were divided into 3 treatment groups (n=8/group): 1) vehicle (HFDV; 1% Na-carboxymethyl cellulose; p.o.); 2) MAC28 (40 mg/kg; p.o.); 3) metformin (300 mg/kg; p.o.; a positive control), the ND-fed rats received a vehicle (NDV). Rats were received their treatment for 4 weeks. Then, LV function and heart rate variability (HRV) were examined, and the heart was removed to determinecardiac malondialdehyde (MDA), cardiac inflammation (TNF-α) and mitochondrial function. Results HFD-induced prediabetes, together with depressed HRV and %LV ejection fraction (LVEF) (Fig. 1A). Moreover, cardiac oxidative stress and inflammation overproduction, and cardiac mitochondrial dysfunction was also observed, shown by elevated cardiac MDA, cardiac TNF-α protein levels, and mitochondrial ROS levels, mitochondrial depolarization and swelling (Fig. 1B). Notably, treatment with MAC28 effectively improved HRV and %LVEF and HRV (Fig. 1A), compared to HFDV group. Moreover, MAC28 significantly reduced cardiac MDA levels, cardiac TNF-α protein levels and cardiac mitochondrial dysfunction in HFD-induced prediabetic rats (Fig. 1B). These beneficial effects were also observed in metformin-treated rats (Fig. 1A, B). Conclusion MAC28 exerts cardioprotection in prediabetic rats by reducing cardiac oxidative stress, inflammation, and mitochondrial dysfunction, leading to restoring LV function. Funding Acknowledgement Type of funding sources: Public grant(s) – National budget only. Main funding source(s): This work was supported by the Thailand Science Research and Innovation