A H 2 S Donor GYY4137 Exacerbates Cisplatin-Induced Nephrotoxicity in Mice

Abstract

Accumulating evidence demonstrated that hydrogen sulfide (H2S) is highly involved in inflammation, oxidative stress, and apoptosis and contributes to the pathogenesis of kidney diseases. However, the role of H2S in cisplatin nephrotoxicity is still debatable. Here we investigated the effect of GYY4137, a novel slow-releasing H2S donor, on cisplatin nephrotoxicity in mice. Male C57BL/6 mice were pretreated with GYY4137 for 72 h prior to cisplatin injection. After cisplatin treatment for 72 h, mice developed obvious renal dysfunction and kidney injury as evidenced by elevated blood urea nitrogen (BUN) and histological damage. Consistently, these mice also showed increased proinflammatory cytokines such as TNF-α, IL-6, and IL-1β in circulation and/or kidney tissues. Meanwhile, circulating thiobarbituric aid-reactive substances (TBARS) and renal apoptotic indices including caspase-3, Bak, and Bax were all elevated. However, application of GYY4137 further aggravated renal dysfunction and kidney structural injury in line with promoted inflammation, oxidative stress, and apoptotic response following cisplatin treatment. Taken together, our results suggested that GYY4137 exacerbated cisplatin-induced nephrotoxicity in mice possibly through promoting inflammation, oxidative stress, and apoptotic response.

Figure 1

Effects of GYY4137 on renal function and tubular injury following cisplatin treatment. (a) Representative images of periodic acid-Schiff staining (×400) of kidneys. (b) Tubular injury score. (c) BUN levels. CTR: control, n = 5; cp: cisplatin, n = 10; and cp + GYY4137: cisplatin + GYY4137, n = 10. Data are means ± SE.

Figure 2

Effects of GYY4137 on cisplatin-induced inflammatory response. (a) Enzyme-linked immunosorbent assay analysis of circulating TNF-α. (b) qRT-PCR analysis of renal TNF-α mRNA expression. (c) qRT-PCR analysis of renal IL-6 mRNA expression. (d) qRT-PCR analysis of renal IL-1β mRNA expression. CTR: control, n = 5; cp: cisplatin, n = 10; and cp + GYY4137: cisplatin + GYY4137, n = 10. Data are means ± SE.

Figure 3

Effects of GYY4137 on cisplatin-induced oxidative stress. (a) Measurement of circulating thiobarbituric acid-reactive substances (TBARS) levels. (b) qRT-PCR analysis of renal SOD3 mRNA expression. (c) qRT-PCR analysis of renal SOD1 mRNA expression. (d) qRT-PCR analysis of renal SOD2 mRNA expression. CTR: control, n = 5; cp: cisplatin, n = 10; and cp + GYY4137: cisplatin + GYY4137, n = 10. Data are means ± SE.

Figure 4

Effects of GYY4137 on cisplatin-induced apoptotic response. (a) qRT-PCR analysis of caspase-3 mRNA expression in kidney. (b) qRT-PCR analysis of Bak mRNA expression in kidney. (c) qRT-PCR analysis of Bax mRNA expression in kidney. CTR: control, n = 5; cp: cisplatin, n = 10; and cp + GYY4137: cisplatin + GYY4137, n = 10. Data are means ± SE.

Figure 5

Effects of GYY4137 on CBS and CSE expression after cisplatin treatment. (a) Western blot analyses of CBS and β-actin in kidney. (b) Densitometry of CBS. A densitometric ratio between the densitometry of CBS and β-actin was calculated, and data are expressed in comparison with the controls. (c) qRT-PCR analysis of CBS. (d) qRT-PCR analysis of CSE. CTR: control, n = 5; cp: cisplatin, n = 10; and cp + GYY4137: cisplatin + GYY4137, n = 10. Data are means ± SE.

Figure 6

Effects of GYY4137 alone on renal morphology and function. (a) Representative images of periodic acid-Schiff staining (×400) of kidneys. (b) Tubular injury score. (c) Plasma creatinine levels. n = 5 in each group. Data are means ± SE.