Ameliorative effect of naringin in acetaminophen-induced hepatic and renal toxicity in laboratory rats: role of FXR and KIM-1

Figures & data

Table 1. Effect of naringin treatment on body weight, relative liver weight, relative spleen weight, albumin, blood urea nitrogen, serum creatinine and uric acid in APAP-induced toxicity in rats.

Treatment	Body weight (BW) (gm)	Relative liver weight	Relative spleen weight	Albumin (mg %)	BUN (mg/dl)	Serum creatinine (mg/dl)	Uric acid (mg %)
Normal	232.50 ± 1.33	0.017 ± 0.001	0.36 ± 0.07	3.21 ± 0.34	7.25 ± 1.50	1.41 ± 0.15	2.24 ± 0.16
APAP control	189.20 ± 6.33^#	0.028 ± 0.002^#	0.89 ± 0.04^#	0.61 ± 0.20^#	24.31 ± 0.73^#	7.27 ± 0.66^#	0.55 ± 0.04^#
Silymarin (25)	226.83 ± 2.89*^,^$	0.024 ± 0.001*^,^$	0.42 ± 0.03*^,^$	2.07 ± 0.32*^,^$	10.63 ± 2.31*^,^$	2.98 ± 0.64*	1.84 ± 0.26*^,^$
Naringin (20)	204.16 ± 1.90	0.027 ± 0.002	0.85 ± 0.06	0.93 ± 0.24	20.43 ± 1.45	4.79 ± 0.75	0.71 ± 0.07
Naringin (40)	220.83 ± 9.10*^,^$	0.024 ± 0.001*^,^$	0.51 ± 0.05*^,^$	1.20 ± 0.26	14.30 ± 0.46*^,^$	3.83 ± 0.45*^,^$	1.70 ± 0.31*^,^$
Naringin (80)	227.33 ± 4.25*^,^$	0.022 ± 0.002*^,^$	0.41 ± 0.03*^,^$	2.39 ± 0.27*^,^$	6.95 ± 1.22*^,^$	3.27 ± 0.60*^,^$	1.74 ± 0.16*^,^$
Per se	228.00 ± 1.34	0.019 ± 0.001	0.35 ± 0.04	2.90 ± 0.26	6.95 ± 1.22	1.47 ± 0.38	1.85 ± 0.17

Notes: Data are expressed as mean ± SEM (n = 6) and analyzed by one-way ANOVA followed by Tukey’s multiple range test for each parameter separately.

*p < 0.05 as compared to APAP group,

# p < 0.05 as compared to normal group and

$ p < 0.05 as compared to one another group.

APAP: acetaminophen; BUN: blood urea nitrogen.

Table 2. Effect of naringin treatment on alkaline phosphatase, total bilirubin, direct bilirubin, AST and ALT in APAP-induced toxicity in rats.

Treatment	Alkaline phosphatase (IU/I)	Total bilirubin (mg %)	Direct bilirubin (mg %)	AST (IU/I)	ALT (IU/I)
Normal	54.62 ± 8.51	0.10 ± 0.01	0.19 ± 0.03	130.5 ± 16.33	38.52 ± 6.65
APAP control	284.0 ± 38.44^#	0.39 ± 0.04^#	0.67 ± 0.06^#	360.1 ± 16.73^#	145.6 ± 10.48^#
Silymarin (25)	131.3 ± 8.98*^,^$	0.12 ± 0.01*^,^$	0.26 ± 0.03*	175.4 ± 20.43*^,^$	48.32 ± 10.38*^,^$
Naringin (20)	210.5 ± 26.65	0.33 ± 0.05	0.51 ± 0.11	290 ± 22.11	142.6 ± 12.36
Naringin (40)	198.4 ± 28.51	0.23 ± 0.02*	0.33 ± 0.08*	248.3 ± 31.16*	96.4 ± 16.68*
Naringin (80)	96.92 ± 9.68*^,^$	0.17 ± 0.04*	0.26 ± 0.04*	177.1 ± 32.26*^,^$	57.93 ± 4.68*^,^$
Per se	49.72 ± 3.57	0.13 ± 0.02	0.22 ± 0.04	155.6 ± 21.63	46.64 ± 10.60

Notes: Data are expressed as mean ± SEM (n = 6) and analyzed by one-way ANOVA followed by Tukey’s multiple range test for each parameter separately.

*p < 0.05 as compared to APAP group,

# p < 0.05 as compared to normal group and

$ p < 0.05 as compared to one another group.

APAP: acetaminophen; AST: aspartate transaminase; ALT: alanine transaminase.

Table 3. Effect of naringin treatment on cholesterol, HDL, LDL, LDH and triglyceride in APAP-induced toxicity in rats.

Treatment	Cholesterol (mg %)	HDL (mg %)	LDL (mg %)	LDH (mg %)	Triglyceride (mg %)
Normal	19.14 ± 3.20	60.94 ± 1.06	1.85 ± 0.34	514.3 ± 120.7	60.45 ± 14.86
APAP control	67.24 ± 8.68^#	21.03 ± 3.41^#	5.09 ± 0.516^#	3377 ± 213.3^#	178.40 ± 19.24^#
Silymarin (25)	33.61 ± 2.48*	26.18 ± 3.66	1.84 ± 0.48*^,^$	680.7 ± 151.0*^,^$	96.21 ± 29.06*^,^$
Naringin (20)	61.74 ± 4.54	29.93 ± 2.01	3.21 ± 0.50	3153 ± 406.3	172.40 ± 17.15
Naringin (40)	47.9 ± 5.01*	44.87 ± 4.76*^,^$	2.21 ± 0.37*	2349 ± 406.3*	107.60 ± 13.84*
Naringin (80)	27.97 ± 4.08*^,^$	55.57 ± 3.21*^,^$	2.14 ± 0.43*	1449 ± 132.2*^,^$	76.37 ± 9.027*^,^$
Per se	27.87 ± 2.28	58.48 ± 4.34	1.78 ± 0.41	647.3 ± 145.9	72.44 ± 10.03

Notes: Data are expressed as mean ± SEM (n = 6) and analyzed by one-way ANOVA followed by Tukey’s multiple range test for each parameter separately.

*p < 0.05 as compared to APAP group,

# p < 0.05 as compared to normal group and

$ p < 0.05 as compared to one another group.

APAP: acetaminophen; HDL: high-density lipoprotein; LDL: low-density lipoprotein; LDH: lactate dehydrogenase; VLDL: very low-density lipoprotein.

Figure 1. Effect of naringin treatment on hepatic and renal SOD (A), GSH (B), MDA (C) and NO (D) in APAP-induced toxicity in rats. Data are expressed as mean ± SEM (n = 6) and analyzed by one-way ANOVA followed by Tukey’s multiple range test for each parameter separately. *p < 0.05 as compared to APAP group, ^#p < 0.05 as compared to normal group and ^$p < 0.05 as compared to one another group. APAP: acetaminophen; N (20): naringin (20 mg/kg) treated; N (40): naringin (40 mg/kg) treated; N (80): naringin (80 mg/kg) treated; SOD: superoxide dismutase; GSH: glutathione; MDA: malondialdehyde; NO: nitric oxide.

Figure 2. Effect of naringin treatment on liver FXR mRNA expression (A), renal KIM-1 mRNA expression (B) in APAP-induced toxicity in rats. Data are expressed as mean ± SEM (n = 6) and analyzed by one-way ANOVA followed by Tukey’s multiple range test for each parameter separately. *p < 0.05 as compared to APAP group, ^#p < 0.05 as compared to normal group and ^$p < 0.05 as compared to one another group. APAP: acetaminophen; FXR: farnesoid X receptor; KIM-1: kidney injury molecule-1.

Figure 3. Effect of naringin treatment on APAP-induced pathological alteration in rat liver. Photomicrograph of sections of liver of normal (A), APAP controls (B), silymarin (25 mg/kg) treated (C), naringin (40 mg/kg) treated (D), naringin (80 mg/kg) treated (E) and per se treated (F) rats. Inflammatory infiltration (black arrow), edema (yellow arrow), congestion (green arrow) and necrosis (red arrow). H&E staining at 40× and 100 × (inset).

Table 4. Effect of naringin treatment on APAP-induced pathological alteration in rat hepatic and renal histology.

Treatment	Normal	APAP control	Silymarin (25)	Naringin (20)	Naringin (40)	Naringin (80)	Per se
Hepatic changes
Vesicular fat	0	++	+	++	++	+	0
Inflammatory infiltration	0	++++	+	++++	+++	++	0
Plasma cells	0	+++	+	+++	++	++	0
Congestion	+	+++	++	++	++	+	0
Edema	0	+++	++	++	+	+	0
Vacuolization	0	+++	+	++	+	0	+
Pyknosis	0	+++	+	++	+	+	0
Necrosis	0	++++	0	+++	+	+	0
Renal changes
Glomerular hypertrophy	0	++++	++	+++	++	+	0
Inflammatory infiltration	0	+++	+	++	++	+	0
Congestion	0	++++	+	+++	+	+	+
Edema	+	+++	++	++	+	+	0
Pyknosis	0	++	+	++	+	+	+
Necrosis	0	+++	0	++	+	+	0

Notes: APAP: acetaminophen; 0: no abnormality detected; +: damage/active changes up to <25%; ++: damage/active changes up to <50%; +++: damage/active changes up to <75%; ++++: damage/active changes up to >75%.

Figure 4. Effect of naringin treatment on APAP-induced pathological alteration in rat kidney. Photomicrograph of sections of kidney of normal (A), APAP controls (B), silymarin (25 mg/kg) treated (C), naringin (40 mg/kg) treated (D), naringin (80 mg/kg) treated (E) and per se treated (F) rats. Glomerular hypertrophy (blue arrow), inflammatory infiltration (black arrow), edema (yellow arrow), congestion (green arrow) and necrosis (red arrow). H&E staining at 40× and 100× (inset).