Abstract

 
 
 
 Purpose: To determine the potential effect of ruscogenin in cisplatin-induced nephrotoxicity.
 Methods: Rat renal tubular epithelial cells (NRK-52E) were treated with 50 μM cisplatin to establish an in vitro cell model of nephrotoxicity. Cytotoxicity was assessed by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, flow cytometry, and western blot. Different concentrations of ruscogenin (2.5, 5, and 10 μM) were incubated with cisplatin-treated NRK-52E cells. Alterations in the nod-like receptor family, the pyrin domain-containing protein (NLRP3) inflammasome, toll-like receptor 4 (TLR4)/nuclear factor kappa B (NF-κB), and nuclear factor erythropoietin-2-related factor 2 (Nrf2)/heme oxygenase 1 (HO-1) components were determined using western blot. Flow cytometry was also used to investigate the levels of reactive oxygen species (ROS).
 Results: Ruscogenin significantly increased cell viability (p < 0.01) and suppressed apoptosis of NRK- 52E cells (p < 0.01), attenuating cisplatin-induced cytotoxicity. The NLRP3 inflammasome was activated in cisplatin-treated NRK-52E cells with enhanced NLRP3, interleukin 1 beta, and cleaved caspase-1; however, ruscogenin significantly decreased the expression of NLRP3 inflammasome components (p < 0.01). Ruscogenin attenuated cisplatin-induced expression of TLR4, myeloid differentiation primary response 88, and NF-κB. Further, cisplatin induction enhanced ROS formation, with increased malondialdehyde and decreased glutathione reductase and catalase levels. Ruscogenin attenuated cisplatin-induced ROS accumulation in NRK-52E cells through up-regulation of Nrf2 and HO-1.
 Conclusion: Ruscogenin protects against cisplatin-induced apoptosis, inflammation, and oxidative stress in renal tubular epithelial cells via suppression of TLR4/NF-κB activation and promotion of Nrf2/HO-1 activation. Therefore, ruscogenin provides a potential therapeutic strategy for mitigating cisplatin-induced nephrotoxicity.
 
 
 
Highlights
Acute kidney injury, characterized by a loss of kidney function, is a common complication in hospitalized patients [1]
Ruscogenin showed an anti-inflammatory effect on cisplatintreated renal tubular epithelial cells through a dose-dependent reduction of NLRP3, IL-1β, and cleaved caspase-1 (Figure 2)
Cell viability of NRK-52E cells was decreased by cisplatin (Figure 1A), and apoptosis was promoted by cisplatin (Figure 1B)
Summary
Acute kidney injury, characterized by a loss of kidney function, is a common complication in hospitalized patients [1]. Cisplatin treatment induces proximal tubular epithelial cell apoptosis, inflammation, and oxidative stress to aggravate the renal injury [6]. The effect of ruscogenin on cisplatin-induced acute kidney injury has not been reported. The protective effect of ruscogenin on cisplatintreated rat renal proximal tubular epithelial cells (NRK-52E) was assessed. Rat renal proximal tubular epithelial cells (NRK52E) were plated in a 96-well plate and incubated with cisplatin and/or ruscogenin for 24 h. NRK-52E cells were harvested and suspended in 100 μL Annexin V-binding buffer (Thermo Fisher Scientific). Cell apoptosis in NRK-52E cells was assessed using an AttuneTM Flow Cytometer (Thermo Fisher Scientific). NRK-52E cells, after cisplatin and/or ruscogenin treatment, were harvested and suspended in 100 μL serum-free medium with 10 μM DCFH-DA (Cellular ROS Assay Kit; Abcam, Cambridge, MA, USA) for 30 min. A p value less than 0.05 was considered as statistically significant
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
