Dapagliflozin Alleviates Renal Fibrosis by Inhibiting RIP1-RIP3-MLKL-Mediated Necroinflammation in Unilateral Ureteral Obstruction

Abstract

Dapagliflozin, a sodium-glucose cotransporter-2 inhibitor, offers renoprotection in diabetes. However, potential for use in nondiabetic kidney disease remains unknown. Herein, we assessed whether dapagliflozin alleviates renal fibrosis by interfering with necroinflammation in a rat model of unilateral ureteral obstruction (UUO) and in vitro. After induction of UUO, rats were administered dapagliflozin daily for seven consecutive days. UUO induced significant renal tubular necrosis and overexpression of RIP1-RIP3-MLKL axis proteins; these coincided with NLRP3 inflammasome activation, and subsequent development of renal fibrosis. Oxidative stress caused by UUO is tightly associated with endoplasmic reticulum stress and mitochondrial dysfunction, leading to apoptotic cell death through Wnt3α/β-catenin/GSK-3β signaling; all of which were abolished by both dapagliflozin and specific RIP inhibitors (necrostatin-1 and GSK872). In H2O2-treated HK-2 cells, dapagliflozin and RIP inhibitors suppressed overexpression of RIP1-RIP3-MLKL proteins and pyroptosis-related cytokines, decreased intracellular reactive oxygen species production and apoptotic cell death, whereas cell viability was improved. Moreover, activated Wnt3α/β-catenin/GSK-3β signaling was inhibited by dapagliflozin and Wnt/β-catenin inhibitor ICG-001. Our findings suggest that dapagliflozin ameliorates renal fibrosis by inhibiting RIP1-RIP3-MLKL-mediated necroinflammation via Wnt3α/β-catenin/GSK-3β signaling in UUO.

Dapagliflozin alleviates renal fibrosis in a mouse model of adenine-induced renal injury by inhibiting TGF-β1/MAPK mediated mitochondrial damage

“…This is partly due to the lack of effective drugs. DN as an important cause of CKD, and some drugs treated with diabetes like SGLT2 inhibitors have shown renal protective effects ( Sharma et al, 2017 ; Kelly et al, 2019 ), as a typical drug of SGLT2 inhibitors, dapagliflozin’s renal protective effects on CKD have been demonstrated in patients with and without diabetes ( Kurata and Nangaku, 2022 ) , and it has also displayed antirenal fibrosis activities in animal models induced by unilateral ureter obstruction and alcohol damage ( Wu et al, 2021 ; Xuan et al, 2021 ; Liu et al, 2022 ). In this study, we extended this model to the setting of renal fibrosis induced by a 0.2% adenine diet.…”

“…Inflammation and oxidative stress were critical aspects of renal fibrosis ( Meng et al, 2014 ; Rayego-Mateos and Valdivielso, 2020 ). Dapagliflozin reduces inflammation and oxidative stress in the kidneys in UUO animal models ( Xuan et al, 2021 ; Liu et al, 2022 ). Our study also showed that dapagliflozin increased content of GSH, improved activity of SOD, and decreased the content of MDA in 0.2% adenine diet-fed mice.…”

“…The sections were stained with uranyl acetate and lead citrate, dried. Using transmission electron microscope (Hitachi; HT7800/HT7700), the number and size of mitochondria were measured in 20 random unoverlapped proximal tubular cells of each sample ( Xuan et al, 2021 ).…”

“…Due to its protective effect in the kidneys of patients with non-diabetic chronic kidney disease, research into the use of dapagliflozin to treat renal fibrosis is underway. Studies have shown that dapagliflozin can reduce renal fibrosis in alcohol-induced kidney injury and unilateral ureteral obstruction (UUO) animal models ( Wu et al, 2021 ; Xuan et al, 2021 ; Liu et al, 2022 ).…”

Autophagy and necroptosis in cisplatin-induced acute kidney injury: Recent advances regarding their role and therapeutic potential

“…Under the conditions of unilateral ureteral obstruction and renal ischemia/reperfusion injury, tubular necroptosis is markedly upregulated, which in turn promotes NOD-like receptor protein 3 (NLRP3) inflammasome activation leading to renal fibrosis. Whereas genetic or pharmacologic inhibition of the necroptosis axis prevented activation of necroinflammation and subsequent development of renal fibrogenesis ( Chen et al, 2018 ; Jiang et al, 2022 ; Xuan et al, 2022 ), implying a relationship between necroinflammation and renal fibrosis. Most importantly, in the CP-induced AKI model, Landau et al (2019) found that continuous activation of regulated necrosis following CP treatment was shown to be the most important factor that drives the transition and progression of AKI to chronic irreparable kidney disease.…”