Enavogliflozin attenuates early sepsis-associated acute kidney injury in a murine cecal ligation and puncture model

Abstract

Sepsis is often complicated by the onset of acute kidney injury (AKI) through processes involving inflammation, oxidative stress, tubular damage, and cell death. In this study, we investigated whether enavogliflozin might mitigate acute renal damage following Cecal Ligation and Puncture (CLP)-induced sepsis.

Methods. Twenty-four male mice were allocated to the following four experimental groups: sham operation (Sham), CLP, CLP plus dimethyl sulfoxide (DMSO), and CLP plus enavogliflozin. Enavogliflozin was administered intraperitoneally at 1mg/kg, 1h before CLP surgery. At 24 hours after surgery, blood and kidney tissue samples were collected. Serum creatinine was measured to assess kidney function, while renal tissue levels of kidney injury molecule-1 (KIM-1), nuclear factor kappa B (NF-κB), malondialdehyde (MDA), caspase-3, and nuclear factor erythroid 2-related factor 2 (Nrf2) were measured using an enzyme-linked immunosorbent assay (ELISA). We also performed a kidney histopathological assessment using Hematoxylin & Eosin staining and a score assigned for renal damage.

Results. Serum creatinine was significantly elevated in mice undergoing CLP compared to sham controls (2.240 ± 0.2546 mg/dL vs 0.4307 ± 0.05516 mg/dL; P < 0.0001). Administration of enavogliflozin in CLP animals resulted in a decrease in serum creatinine levels (1.117 ± 0.1276 mg/dL vs 2.240 ± 0.2546 mg/dL; P < 0.01). Compared with the CLP group, enavogliflozin significantly reduced renal tissue levels of the tubular injury marker KIM-1 (359.3 ± 12.30 vs. 461.4 ± 18.32 pg/mL; P < 0.001), the inflammatory mediator NF-κB (844.7 ± 20.09 vs. 1429 ± 11.38 pg/mL; P < 0.0001), the oxidative stress marker MDA (204.5 ± 18.79 vs. 320.6 ± 23.51 ng/mL; P < 0.01), and the apoptosis-related protein caspase-3 (7.726 ± 0.4581 vs. 10.21 ± 0.5911 ng/mL; P < 0.05). In contrast, renal Nrf2 levels were significantly higher in enavogliflozin-treated mice than in the CLP group (4383 ± 123.6 vs. 3210 ± 117.6 pg/mL; P < 0.0001), indicating enhancement of the antioxidant response.  Histologically, renal tissue of enavogliflozin-treated mice showed ameliorated injury scores compared to the CLP controls [0.0 (0.0–1.0) vs 3.5 (3.0–4.0); P < 0.05].

Conclusions. Treatment with enavogliflozin attenuates acute CLP-induced renal injury by ameliorating tubular damage, suppressing inflammation and oxidative stress, and promoting an antioxidative cellular response. Further studies are warranted to elucidate the mechanism of action and establish the clinical utility of enavogliflozin administration following sepsis.