Betulinic acid protects against N-nitrosodimethylamine-induced redox imbalance in testes of rats

Figures & data

Table 1. Body weight and relative weight of testes of rats treated with NDMA and BA (alone and in combination).

Grouping	Weights (g)			Weight of testes (g)	Relative weight of testes (% of body weight)
	Initial	Final	Weight gain
Control	136.67 ± 3.50	192.50 ± 5.73	55.83 ± 2.83	2.03 ± 0.13	1.06 ± 0.02
BA	131.00 ± 4.01	195.00 ± 2.25	64.17 ± 4.29^a	2.18 ± 0.19	1.12 ± 0.05^b
NDMA	131.67 ± 3.83	163.33 ± 6.69	31.67 ± 1.69^c	3.06 ± 0.16^c	1.86 ± 0.02^c
BA + NDMA	136.67 ± 5.16	184.17 ± 4.00	47.50 ± 4.05^a	2.10 ± 0.18^a	1.18 ± 0.05^a

Data expressed as mean ± S.D. (n = 6).

^aStatistically different from NDMA (P < 0.05).

^bStatistically similar to [BA + NDMA] (P > 0.05).

^cStatistically different from control (P < 0.05).

BA: betulinic acid; NDMA: N-nitrosodimethylamine.

Table 2. Sperm qualities of rats treated with NDMA and BA (alone and in combination).

Treatments groups	Motility (%)	Live–dead ratio	Sperm volume (ml)	Sperm count (×10^6/ml)	TSA (%)
Control	87.50 ± 6.12	97.00 ± 1.55	5.18 ± 0.04	130.50 ± 8.76	7.48 ± 0.14
BA	83.83 ± 6.65^a	96.50 ± 1.64^a	5.17 ± 0.05^a	112.33 ± 9.24^a	7.80 ± 0.54^a
NDMA	60.11 ± 4.01^b	70.83 ± 9.70^b	5.18 ± 0.04	83.33 ± 8.14^b	12.14 ± 0.42^b
BA + NDMA	85.67 ± 6.06^c	91.83 ± 7.52^c	5.17 ± 0.05	107.33 ± 7.76^c	8.48 ± 0.72^c

Data expressed as mean ± S.D. (n = 6).

^aStatistically similar to [BA + NDMA] (P > 0.05).

^bStatistically different from control (P < 0.05).

^cStatistically different from NDMA (P < 0.05).

TSA: total sperm abnormality; BA: betulinic acid; NDMA: N-nitrosodimethylamine.

Table 3. Follicle-stimulating hormone, testosterone and prolactin levels in the rats treated with NDMA and BA (alone and in combination).

Treatments	FSH	LH	Testosterone
	(pg/ml)			Prolactin
Control	9.25 ± 2.06	5.18 ± 0.94	2.43 ± 0.70	8.35 ± 2.08
BA	8.05 ± 1.41	5.02 ± 0.87^a	2.58 ± 0.73^a,b	8.50 ± 3.11^a
NDMA	5.25 ± 2.63^c	3.64 ± 0.58^c	1.53 ± 0.76^c	9.05 ± 6.95
BA + NDMA	8.25 ± 2.50^b	4.39 ± 0.30^b	2.13 ± 0.75^b	8.75 ± 2.22

Data expressed as mean ± S.D. (n = 6).

^aStatistically similar to [BA+NDMA] (P > 0.05).

^bStatistically different from NDMA (P < 0.05).

^cStatistically different from control (P < 0.05).

BA: betulinic acid; NDMA: N-nitrosodimethylamine; FSH: follicle-stimulating hormone; LH: luteinizing hormone.

Figure 1. The levels of testicular TBARS in rats treated with NMDA and BA (alone and combination).

Figure 2. Activity of SOD in rats treated with NMDA and BA (alone and combination).

Figure 3. The activities of glutathione peroxidase and CAT in rats treated with NMDA and BA (alone and combination).

Figure 4. Levels of testicular GSH in rats treated with NMDA and BA (alone and combination).

Figure 5. Activity of testicular GST in rats treated with NMDA and BA (alone and combination).

Table 4. Testicular TC, TG and PL levels in the rats treated with NDMA and BA (alone and in combination).

Treatments	TC (mM/l)	TG (mM/l)	PL (mM/l)	TC/PL
Control	0.37 ± 0.09	0.45 ± 0.30	1.80 ± 0.37	0.17
BA	0.35 ± 0.12^a	0.66 ± 0.52^a	1.95 ± 0.43^a	0.18
NDMA	0.52 ± 0.18^b	1.12 ± 0.39^b	1.02 ± 0.28^b	0.51^b
BA + NDMA	0.41 ± 0.33^c	0.72 ± 0.12^c	1.75 ± 0.57	0.25^c

Data expressed as mean ± S.D. (n = 6).

^aStatistically similar to [BA + NDMA] (P > 0.001).

^bStatistically different from control (P < 0.05).

^cStatistically different from NDMA (P < 0.05).

TC: total cholesterol; TG: triglycerides; PL: phospholipids; BA: betulinic acid; NDMA: N-nitrosodimethylamine.

Figure 6. Representative photomicrographs of testes from control and NDMA-treated rats. Control testes (A) showed normal morphology. Treatment-related lesions such as mild congestion, degeneration of the seminiferous tubules and distortion in the testes of NDMA-treated rats (C) were reduced in [NDMA+BA]-treated rats (D), while photomicrographs from BA-treated rats (B) appeared similar to the control (A). BA: betulinic acid; NDMA: N-nitrosodimethylamine.

Table 5. Activities of phase 1 enzymes [aniline hydroxylase (AnH) and aminopyrine-N-demethylase (AmD)] and phase II enzyme (UDPGT) in the testis of rats treated with NDMA and BA (alone and in combination).

Treatments groups	AnH	AmD	UDPGT
	(μmol/mg protein)		(mmol/mg protein)
Control	0.398 ± 0.59	0.530 ± 0.01	0.12 ± 0.02
BA	1.014 ± 098^a	0.485 ± 0.01^a	0.14 ± 0.03^a
NDMA	1.694 ± 0.86^b	0.840 ± 0.08^b	0.07 ± 0.007^b
BA + NDMA	0.910 ± 0.21^c	0.612 ± 0.03^c	0.10 ± 0.03^c

Data expressed as mean ± S.D. (n = 6).

^aStatistically similar to [BA + NDMA] (P > 0.001).

^bStatistically different from control (P < 0.05).

^cStatistically different from NDMA (P < 0.05).

BA: betulinic acid; NDMA: N-nitrosodimethylamine.