Effect of Nigella sativa (black seeds) against methotrexate-induced nephrotoxicity in mice

Abstract

Aim: To evaluate the protective effect of Nigella sativa (NS) against nephrotoxicity of methotrexate (MTX) in mice.

Materials and methods: Four groups of Swiss albino male mice, eight in each group were used. The study was carried on between October 2014 and April 2015. Group 1 (control) were administered 0.3 ml distilled water orally daily for 21 days and injected with normal saline (0.25 ml) IP weekly. Group 2 (MTX group) were treated with MTX, 10 mg/kg IP weekly, while Group 3 were treated with 0.125 ml of NS oil by mouth daily and injected with normal saline (0.25 ml) IP weekly. Group 4 received 0.125 ml of NS oil by mouth daily and injected with 10 mg/kg MTX IP weekly. Oral treatments were administered using a special curved smooth tip nontraumatic metal needle and IP injections were given for 3 weeks at days 7, 14 and 21. Animals were sacrificed at day 23. Malondialdehyde (MDA) and glutathione (GSH) measurements were performed on kidney homogenate. Histopathology of the kidneys were prepared and examined.

Results: MTX has resulted in a small elevation in MDA and reduction in GSH levels in kidney homogenate which was returned back to control values when NS and MTX were administered in combination. Statistical significance was achieved with elevation of GSH by MTX and NS compared to MTX alone. MTX caused histopathological changes suggesting nephrotoxicity in 6 animals out of 8, while no changes were found in all animals treated with MTX and NS.

Conclusion: NS is protective against MTX-induced nephrotoxicity.

Keywords: Antioxidant; Nigella sativa; kidney toxicity; methotrexate.

Figure 1

(a) Changes in animal body weights between day 1 (before treatment) and after 23 days of treatment with MTX, NS and the combination, (b) the ratio of kidney weights to animal weights. C: Control, MTX: Methotrexate, NS: Nigella sativa, NS+MTX: Combination of NS and MTX. *: Significantly different from the control (P=0.05)

Figure 2

Renal tissue from a mouse treated with normal saline, (a) Nigella sativa, (b) histopathological features revealed normal glomeruli and tubules (×40 hematoxylin and eosin)

Figure 3

Renal tissue of a mouse treated with methotrexate (MTX), (a) showed severe focal degeneration of tubular epithelial cells (×40, hematoxylin and eosin [H&E]), (b) it is a renal tissue from a mouse treated with the combination Nigella sativa and MTX. Normal glomeruli and tubules are seen (×40, H&E)