Protective Effect of Croton macrostachyus (Euphorbiaceae) Stem Bark on Cyclophosphamide-Induced Nephrotoxicity in Rats

Figures & data

Table 1 Grouping and Dosing of Rats Used in This Experiment

Treatment Groups (Crude Extract and Aqueous Fraction)	Dose
Normal control (Normal saline)	0.5 mL/100 gm
Cyclophosphamide (CP) control	200 mg/kg
Crude extract (CE100)	100 mg/kg
Crude extract (CE200)	200 mg/kg
Crude extract (CE400)	400 mg/kg
Aqueous fraction (AQ100)	100 mg/kg
Aqueous fraction (AQ200)	200 mg/kg
Treatment Groups (Ethyl Acetate Fraction)	Dose
Normal control (2% DMSO)	0.5 mL/100 gm
Cyclophosphamide (CP) control	200 mg/kg
Ethyl acetate fraction (EA100)	100 mg/kg
Ethyl acetate fraction (EA200)	200 mg/kg

Abbreviations: AQ, aqueous fraction; CE, crude extract; CP, cyclophosphamide; DMSO, dimethyl sulfoxide; EA, ethyl acetate fraction.

Table 2 Percentage Yield of Crude Extract and Solvent Fractions of Croton macrostachyus

Extract/Fraction	Actual Mass (g)	Percentage Yield (w/w)
Crude extract	101.5	6.68%
Ethyl acetate	11.4	17.53%
Aqueous	36.7	56.46%

Table 3 Effect of Crude Extract and Solvent Fractions of Croton macrostachyus on Kidney Weight and Kidney-to-Body Weight Ratio

Group	Dose	Kidney Weight (gm)	Kidney to Body Wt. Ratio (x10^−3)
NC	NS (0.5 mL/100 gm)	0.88±0.17	2.71±0.06
CP control	CP 200 mg/kg	1.02±0.01**	4.85±0.04***
CE	CE 100 mg/kg	0.94±0.02	3.46±0.04^###
	CE 200 mg/kg	0.99±0.02	3.87±0.02^#
	CE 400 mg/kg	0.90±0.01^##	3.84±0.02^##
AF	AQ 100 mg/kg	0.90±0.03^#	4.28±0.15
	AQ 200 mg/kg	0.86±0.02^##	3.99±0.10^#
NC	2% DMSO (0.5 mL/100 gm)	0.48±0.04	1.79±0.13
CP control	CP 200 mg/kg	0.73±0.03**	4.83±0.28***
EA	EA 100 mg/kg	0.67±0.03	3.57±0.11^###
	EA 200 mg/kg	0.66±0.03	3.48±0.07^###

Notes: Results were expressed as mean ± SEM (n = 6). **_P < 0.01, ***_P < 0.001 when compared with normal control; and ^#_P < 0.05, ^##_P < 0.01, ^###_P < 0.001 when compared with CP control.

Abbreviations: AF, aqueous fraction; CE, crude extract; DMSO, dimethyl sulfoxide; EA, ethyl acetate fraction; CP, cyclophosphamide; NC, normal control; NS, normal saline.

Table 4 Effect of Crude Extract and Aqueous Fraction of Croton macrostachyus on Renal Biomarkers

Group	Dose	Serum Creatinine (mg/dl)	BUN (mg/dl)
NC	NS (0.5 mL/100 gm)	0.55 ± 0.05	32.83 ± 3.72
CP control	CP 200 mg/kg	1.56 ± 0.08	152.00 ± 8.50***
CE	CE 100 mg/kg	0.65 ± 0.05	26.75 ± 2.75^###
	CE 200 mg/kg	0.78 ± 0.09	41.60 ± 5.07^###
	CE 400 mg/kg	0.60 ± 0.03	29.60 ± 3.29^###
AF	AQ 100 mg/kg	0.78 ± 0.03	25.50 ± 2.48^###
	AQ 200 mg/kg	0.76 ± 0.03	23.00 ± 1.61^###
NC	DMSO (0.5 mL/100 gm)	0.65 ± 0.08	26.00 ± 3.35
CP control	CP 200 mg/kg	1.60 ± 0.14***	51.00 ± 2.67***
EA	EA 100 mg/kg	1.13 ± 0.03^#	30.17 ± 1.62^###
	EA 200 mg/kg	1.01 ± 0.05^##	25.17 ± 1.08^###

Notes: Results were expressed as mean ± SEM (n = 6). ***_P < 0.001 when compared with normal control; and ^#_P < 0.05, ^##_P < 0.01, ^###_P < 0.001 when compared with CP control.

Abbreviations: AF, aqueous fraction; CE, crude extract; DMSO, dimethyl sulfoxide; EA, ethyl acetate fraction; CP, cyclophosphamide; NC, normal control; NS, normal saline.

Figure 1 Histopathological changes in the kidney tissue treated with crude extract and aqueous fraction. (A) Normal control group (normal structure), (B) Cyclophosphamide control group (multifocal necrosis), (C) Crude extract 100 mg/kg (damaged glomeruli), (D) Crude extract 200 mg/kg (nearly normal glomeruli), (E) Crude extract 400 mg/kg (pigment laden macrophage), (F) Aqueous fraction 100 mg/kg (increased inflammatory cells), (G) Aqueous fraction 200 mg/kg (increased inflammatory cells).

Figure 2 Histopathological changes in the kidney tissue treated with ethyl acetate fraction. (A) Normal control group (normal structures), (B) Cyclophosphamide control group (necrosis), (C) Ethyl acetate fraction 100 mg/kg (necrosis), (D) Ethyl acetate fraction 200 mg/kg (inflammatory cells and congestion).