Sevoflurane ameliorates LPS-induced inflammatory injury of HK-2 cells through Sirtuin1/NF-κB pathway

Abstract

The anesthetic sevoflurane (SEV) has been shown to protect against organ’s injury during sepsis. The present study intended to uncover the protective effects of SEV on sepsis-induced acute kidney injury (SI-AKI) and its possible mechanism. Human renal tubular epithelial cell HK-2 was treated with 10 μg/mL lipopolysaccharide (LPS) to construct SI-AKI cell model. LPS-induced HK-2 cells were pretreated with SEV in the absence or presence of EX527, an inhibitor of Sirtuin 1 (SIRT1), after which were the detection of cell viability, lactate dehydrogenase (LDH) release, apoptosis, inflammation, and oxidative stress. Our results demonstrated that LPS caused decreased cell viability, increased LDH release, improved cell apoptosis along with decreased expression of Bcl2 and enhanced expressions of Bax, cleaved PARP and cleaved caspase, enhanced production, and protein expressions of TNF-α, IL-6, and IL-1β, increased generation of reactive oxygen species (ROS) and malondialdehyde (MDA), but contributed to declined activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px). LPS inhibited SIRT1 and IκBα expressions but upregulated p-NF-κB p65 and acetyl-p53 expressions as well. However, SEV pretreatment abolished all above-mentioned effects of LPS on HK-2 cells, while EX527 significantly reversed the effects of SEV. In conclusion, SEV effectively protected HK-2 cells against LPS-induced apoptosis, inflammation, and oxidative stress, and these effects may depend on the increase of SIRT1 expression, thereby inactivating NF-κB signaling.

KDM2B regulates inflammation and oxidative stress of sepsis via targeting NF‐κB and AP‐1 pathways

“…The reported incidence of AKI in septic patients ranges from 22% to 53%. Previous studies have shown that damage to RTEC is a main underlying cause of AKI 2 . KDM2B is involved in numerous pathological processes, for instance, cell senescence and tumor development 5 .…”

“…HK‐2 cells which are epithelial cell line from normal adult human kidney were maintained in DMEM added with 10% FBS in a 37°C incubator supplied with 5% CO 2 . For the AKI model establishment, the HK‐2 cells were treated with 10 μg/mL LPS for 24 h to induce sepsis‐associated AKI 2,9 . The LPS‐induced HK‐2 cells were treated with 1 μM SR11302 as an AP‐1 inhibitor or JSH‐23 as an NF‐κB inhibitor, 10 48 h later, the supernatant was used for cytokines analysis and the cells were collected for western blots analysis.…”

“…Twenty‐four hours later, the cells were replaced with new medium supplemented with 10 μL CCK‐8 per well and cultured for another 5 days. The absorbance was analyzed at 450 nm wavelength, and the cell viability ratio was calculated 2 …”

“…Even if AKI patients survive, the likelihood of chronic kidney disease is greatly increased. It has been reported that damage to renal tubular epithelial cells (RTEC) is the main underlying cause of AKI 2 . However, the molecular mechanisms of RTEC injury in AKI are not fully elucidated.…”

“…It has been reported that damage to renal tubular epithelial cells (RTEC) is the main underlying cause of AKI. 2 However, the molecular mechanisms of RTEC injury in AKI are not fully elucidated.…”