Aqueous extract of Boerhaavia diffusa root ameliorates ethylene glycol-induced hyperoxaluric oxidative stress and renal injury in rat kidney

Figures & data

Figure 1.  HPTLC Fingerprint of aqueous extract of roots of Boerhaavia diffusa Linn.

Figure 2.  DPPH free radicals scavenging activity BDE with reference to standard (ascorbic acid).

Table 1.  Effect of BDE treatment on oxalate excretion and renal function in rats.

Observational parameter	Control	Untreated	Treated (BDE 100 mg/kg)	Treated (BDE 200 mg/kg)
Urine volume (mL/24 h)	6.77 ± 0.48	12.15 ± 0.34^a	16.04 ± 0.36^a,d	24.03 ± 1.29^a,c,e
Oxalate (mg/mL)	0.460 ± 0.061	1.852 ± 0.61^a	0.617 ± 0.29^c	0.545 ± 0.049^c
Blood urea nitrogen (mg/dl)	37.21 ± 1.91	50.67 ± 2.28^a	36.13 ± 2.01^c	32.91 ± 1.68^c
Serum creatinine (mg/dl)	0.568 ± 0.025	0.879 ± 0.029^b	0.575 ± 0.059^d	0.357 ± 0.085^c
Creatinine clearance (mg/min)	0.682 ± 0.062	0.325 ± 0.045^a	0.590 ± 0.037^d	0.740 ± 0.025^c

Values are expressed in mean ± SEM (n = 6).

^ap < 0.001 compared with control group.

^bp < 0.01 compared with control group.

^cp < 0.001 compared with untreated group.

^dp < 0.01 compared with untreated group.

^ep < 0.001 compared with treated (BDE 100 mg/kg) group.

Figure 3.  CaOx crystals in the urine of different treatment groups (Leica EZ-4D) at 40 × 10× magnification. (A) Control rats showed very few or no crystals; (B) Untreated rats excreted numerous oval COM and pyramidal shaped COD crystals; (C and D) Treated rats excreted a significantly reduced number of crystals.

Table 2.  Effect of BDE treatment on renal enzymes and other antioxidant markers in rat kidney homogenate analysis.

Antioxidant marker/enzyme	Control	Untreated	Treated (BDE 100 mg/kg)	Treated (BDE 200 mg/kg)
Malondialdehyde (MDA) (µM/mg protein)	6.67 ± 0.03	9.35 ± 0.14^a	5.38 ± 0.02^c	6.47 ± 0.05^c,e
Catalase (µM H2O2 decomposed/min)	68.2 ± 0.32	39.5 ± 0.61^a	53.6 ± 0.52^c	55.4 ± 0.21^c,f
Super oxide dismutase (U/mg protein)	1.623 ± 0.02	0.826 ± 0.08^a	1.441 ± 0.09^c	1.418 ± 0.05^c
Glutathione s-transferase (µM/min/mg protein)	0.721 ± 0.01	0.765 ± 0.03	0.723 ± 0.02^d	0.725 ± 0.03
Reduced glutathione (µM/mg protein)	6.452 ± 0.13	3.78 ± 0.08^a	5.254 ± 0.17^c	5.817 ± 0.15^c,f
Glutathione peroxidase (U/mg protein)	0.720 ± 0.01	0.421 ± 0.03^a	0.678 ± 0.02^a,c	0.690 ± 0.02^c,f
Total protein content (mg/dl)	284 ± 11.2	315 ± 4.28^a	210 ± 7.8^c	261 ± 21.1^c,f

Values are expressed in mean ± SEM (n = 6).

^ap < 0.001 compared with control group.

^bp < 0.01 compared with control group.

^cp < 0.001 compared with untreated group.

^dp < 0.01 compared with untreated group.

^ep < 0.001 compared with treated (BDE 100 mg/kg) group

^fp < 0.001 compared with treated (BDE 100 mg/kg) group.

Table 3.  Histopathological analysis of hyperoxaluria-induced renal damage and crystal deposits.

Observational parameter	Control	Untreated	Treated (BDE 100 mg/kg)	Treated (BDE 200 mg/kg)
Tubular degeneration	−	+++	+	+
Tubular necrosis	−	+++	+	−
Tubular desquamation	−	+++	++	+
Tubular congestion	−	+++	+	+
Glomerular congestion	−	+++	++	−
Glomerular damage	−	+++	++	−
Interstitial edema	−	+++	−	−
Crystal deposits	−	+++	++	+
Gross damage to renal architecture	Normal	Severe	Moderate	Mild

Scores are based on average observations of renal damage observed in minimum 6 different fields per slide.

+ mild, ++ moderate, +++ severe damage and − normal; Similarly for crystal deposits, + rare, ++ few, +++ numerous and− nil.

Figure 4.  Histopathological examination of kidney slides of different treatment groups (Leica EZ-4D) at 10 × 10× magnification. (A) Control rats showing normal renal architecture; (B) Kidney of untreated rats showing polymorphic irregular crystal deposits tubular damage (in figure, black spots inside the rectangular box are crystal deposits); (C) BDE-treated (100 mg/kg) showed few crystal deposits and little dilation of tubules; (D) BDE-treated (200 mg/kg) showed a very few or no crystal deposits and nearly normal renal architecture.