Detam 1 black soybean against cisplatin-induced acute ren failure on rat model via antioxidant, antiinflammatory and antiapoptosis potential

Abstract

Background and aim: Cis-Diamminedichloroplatinum (II) (Cisplatin) is one of the most synthetic anticancer drug but have several adverse effects and one of them is acute ren failure. Cisplatin can induce nephrotoxicity occur via the toxic generation of reactive oxygen species (ROS). Black soybean (Glycine max L. Merr.) has been reported contain high levels of phenolics and anthocyanins that has antioxidant activity. This study aims to determine the effect of ethanol extract of black soybean (EEBS) against cisplatin-induced nephrotoxicity in rats.

Experimental procedure: Cisplatin-induced nephrotoxicity rats treated with EEBS and the blood samples taken on days 0, 9, and 18. The effects of EEBS was evaluated by determining Interferon-γ (IFN-γ), Caspase-3 (Casp-3), and Interleukin-1β (IL-1β) expression using immunohistochemistry (IHC), blood urea nitrogen (BUN), Uric Acid (UA) content and catalase (CAT) content in the blood plasma with colorimetric assay kit.

Results and conclusion: Based on the results, EEBS treatment had successfully reduced pro-inflammatory cytokines IL-1β and IFN-γ, and improved physiological condition by lowering BUN and UA content while increasing CAT activity. No significant effect was found in Casp-3 expression. EEBS has potential to improve acute renal failure condition through inflammatory suppression and renal function improvement.

Keywords: Blood urea nitrogen; Catalase; Fibronectin; Glycine max L. Merr.; Nephrotoxicity.

Graphical abstract

Fig. 1

Chromatogram of Isoflavones as a standard using LC-MS/MS method. (A. Daidzein (RT 2.58), B. Daidzin (RT 1.23), C. Genistein (RT 3.77), D. Genistin (1.43), E. Biochanin A (RT 1.22) and Glycitein (RT 2.72)).

Fig. 2

Chromatogram of EEBS using LC-MS/MS method. EEBS contained A. Daidzein (RT 2.58), B. Daidzin (RT 1.21), C. Genistin (RT 1.43), D. Biochanin A (RT 1.22) and Glycitein (RT 2.72).

Fig. 3

Effect of EEBS towards IFN-γ expression in acute renal disease rat model The slides is seen at 400× (A–D) and 1000× (E–H) magnification. ∗A and E are Negative Control (normal rat); B and F are Positive Control (acute renal disease); C and G are EEBS 300 (Positive Control + EEBS 300 mg/kg BW); D and H are EEBS 600 (Positive Control + EEBS 600 mg/kg BW).

Fig. 4

Effect EEBS toward IFN-γ expression in acute renal disease rat model Data are presented as mean ± SD. NC: negative control (normal rat + untreated EEBS); PC: positive control (Cisplatin-induced rat); EEBS 300: positive control + EEBS 300 mg/kg BW; EEBS 600: positive control + EEBS 600 mg/kg BW. Different letters (a,b,c) show significant difference of IFN-γ positive cell number among treatments based on Tukey HSD post hoc test (p < 0.05).

Fig. 5

Effect EEBS towards Casp-3 expression in acute renal disease rat model The slides is seen at 400× (A–D) and 1000× (E–H) magnification. ∗A and E are Negative Control (normal rat); B and F are Positive Control (acute renal disease); C and G are EEBS 300 (Positive Control + EEBS 300 mg/kg BW); D and H are EEBS 600 (Positive Control + EEBS 600 mg/kg BW).

Fig. 6

Effect EEBS towards Casp-3 expression in acute renal disease rat model Data are presented as mean ± SD. NC: negative control (normal rat + untreated EEBS); PC: positive control (Cisplatin-induced rat); EEBS 300: positive control + EEBS 300 mg/kg BW; EEBS 600: positive control + EEBS 600 mg/kg BW. Different letters (a,ab,b) show significant difference of Casp-3 expression among treatments based on Tukey HSD post hoc test (p < 0.05).

Fig. 7

Effect of EEBS towards IL-1β expression in acute renal disease rat model The slides is seen at 400× (A–D) and 1000× (E–H) magnification. ∗A and E are Negative Control (normal rat); B and F are Positive Control (acute renal disease); C and G are EEBS 300 (Positive Control + EEBS 300 mg/kg BW); D and H are EEBS600 (Positive Control + EEBS 600 mg/kg BW).

Fig. 8

Effect of EEBS towards IL-1β expression in acute renal disease rat model Data are presented as mean ± SD. NC: negative control (normal rat + untreated EEBS); PC: positive control (Cisplatin-induced rat); EEBS 300: positive control + EEBS 300 mg/kg BW; EEBS 600: positive control + EEBS 600 mg/kg BW. Different letters (a,b, c) show significant difference of IL-1β expression among treatments based on Tukey HSD post hoc test (p < 0.05).

Fig. 9

Effect EEBS toward BUN level in acute renal disease rat model Data are presented as mean ± SD. NC: negative control (normal rat + untreated EEBS); PC: positive control (Cisplatin-induced rat); EEBS 300: positive control + EEBS 300 mg/kg BW; EEBS 600: positive control + EEBS 600 mg/kg BW. Different letters (a,b) show significant difference of BUN level among treatments in day 0, different letters (a,b,c,d) show significant difference of BUN content among treatments in day 9, and different letters (a,b,bc,c) show significant difference of BUN content among treatments in day 18. All statistical different was based on Tukey HSD post hoc test (p < 0.05).

Fig. 10

Effect EEBS toward UA level in acute renal disease rat model∗ Data are presented as mean ± SD. NC: negative control (normal rat + untreated EEBS); PC: positive control (Cisplatin-induced rat); EEBS 300: positive control + EEBS 300 mg/kg BW; EEBS 600: positive control + EEBS 600 mg/kg BW. Different letters (a,b) show significant difference of UA content among treatments in day 0, different letters (a,b,ab) show significant difference of UA content among treatments in day 9, and different letters (a,b,c) show significant difference of UA content among treatments in day 18. All statistical different was based on Tukey HSD post hoc test (p < 0.05).

Fig. 11

Effect EEBS toward CAT activity in acute renal disease rat model∗ Data are presented as mean ± SD. NC: negative control (normal rat + untreated EEBS); PC: positive control (Cisplatin-induced rat); EEBS 300: positive control + EEBS 300 mg/kg BW; EEBS 600: positive control + EEBS 600 mg/kg BW. Different letters (a,b) show significant difference of CAT content among treatments in day 0, different letters (a,b,c,d) show significant difference of CAT content among treatments in day 9, and different letters (a,b,c) show significant difference of CAT content among treatments in day 18. All statistical different was based on Tukey HSD post hoc test (p < 0.05).

Fig. 12

Effect EEBS toward renal weight in acute renal disease rat model∗ Data are presented as mean ± SD: NC: negative control (normal rat + untreated EEBS); PC: positive control (Cisplatin-induced rat); EEBS 300: positive control + EEBS 300 mg/kg BW; EEBS 600: positive control + EEBS 600 mg/kg BW. Different letters (a,b) show significant difference of renal weight among treatments in right side, different letters (a,b) show significant difference of renal weight among treatments in left side. All statistical different was based on Tukey HSD post hoc test (p < 0.05).