Synergistic Renoprotective Effect of Melatonin and Zileuton by Inhibition of Ferroptosis via the AKT/mTOR/NRF2 Signaling in Kidney Injury and Fibrosis

Abstract

Kidney diseases pose a tremendous burden to patients and healthcare facilities globally (Luyckx et al., 2018). A recent report indicates that over 700 million individuals are affected by chronic kidney disease (CKD) worldwide (Sundström et al., 2022). According to time frames, acute kidney injury (AKI) and CKD have a substantial impact on mortality and other health-related consequences, including end-stage kidney disease (ESKD) and cardiovascular events (Kellum et al., 2021;Kovesdy, 2022; Matsushita et al., 2022). Despite the recent development of promising therapeutics, such as sodium-glucose co-transporter 2 inhibitors, disease-specific therapies for diverse kidney diseases are still lacking (van Asbeck et al., 2020).Various forms of cell death including apoptosis, necroptosis, autophagy, and ferroptosis are involved in different types of kidney injuries (Priante et al., 2019;Maremonti et al., 2022). Among these cell death processes, non-apoptotic cell death is considered a potential therapeutic target. Regulated necrosis such as ferroptosis and necroptosis is a more immunogenic form of cell death that triggers the innate immune system by 599 According to recent evidence, ferroptosis is a major cell death mechanism in the pathogenesis of kidney injury and fibrosis. Despite the renoprotective effects of classical ferroptosis inhibitors, therapeutic approaches targeting kidney ferroptosis remain limited. In this study, we assessed the renoprotective effects of melatonin and zileuton as a novel therapeutic strategy against ferroptosis-mediated kidney injury and fibrosis. First, we identified RSL3-induced ferroptosis in renal tubular epithelial HK-2 and HKC-8 cells. Lipid peroxidation and cell death induced by RSL3 were synergistically mitigated by the combination of melatonin and zileuton. Combination treatment significantly downregulated the expression of ferroptosis-associated proteins, 4-HNE and HO-1, and upregulated the expression of GPX4. The expression levels of p-AKT and p-mTOR also increased, in addition to that of NRF2 in renal tubular epithelial cells. When melatonin (20 mg/kg) and zileuton (20 mg/kg) were administered to a unilateral ureteral obstruction (UUO) mouse model, the combination significantly reduced tubular injury and fibrosis by decreasing the expression of profibrotic markers, such as α-SMA and fibronectin. More importantly, the combination ameliorated the increase in 4-HNE levels and decreased GPX4 expression in UUO mice. Overall, the combination of melatonin and zileuton was found to effectively ameliorate ferroptosis-related kidney injury by upregulating the AKT/mTOR/ NRF2 signaling pathway, suggesting a promising therapeutic strategy for protection against ferroptosis-mediated kidney injury and fibrosis.

Mechanism of targeting the mTOR pathway to regulate ferroptosis in NSCLC with different EGFR mutations

“…In addition, the effect of GPX4 regulation on the AKT/mTOR pathway was observed and it was found that GPX4 inhibition effectively inhibited the AKT/mTOR pathway in H1975 cells. This may be related to the positive regulatory relationship between AKT/mTOR and GPX4 ( 50 , 51 ). Notably, compared with H1975 and A549 cells, H3255 cells were the most sensitive to the GPX4 inhibitor, RSL3, and the least sensitive to erastin, which may be related to the different mechanisms of these drugs.…”

Sestrin 2 protects human lens epithelial cells from oxidative stress and apoptosis induced by hydrogen peroxide by regulating the mTOR/Nrf2 pathway

“…Jung et al showed that the combination of zileuton and melatonin improved kidney injury by activating the AKT/mTOR/Nrf2 pathway. 59 Zhang et al found that vascular injury was attenuated by irisin, which was associated with the activation of the AKT/mTOR/Nrf2 pathway. 60 In our present study, we found SESN2 could activate mTOR pathway and upregulate Nrf2 expression.…”

Ferroptosis in organ fibrosis: From mechanisms to therapeutic medicines

“…[ 48 ] Our studies confirmed that ACSL4-mediated ferroptosis was involved in UUO and adenine diet-induced kidney fibrosis. [ 6 , 26 ] Other reported signaling pathways include Smad3/ATF3/SLC7A11 signaling, [ 49 ] Akt/GSK-3p/Nrf2 signaling, [ 50 ] XBP1-Hrd1-Nrf2 signaling, [ 51 ] AKT/ mTOR/Nrf2 signaling, [ 52 ] TLR4/Nox4 signaling [ 53 ] and so on ( Figure 3A ).…”