Prevention of arsenic-mediated reproductive toxicity in adult female rats by high protein diet

Figures & data

Table 1. Changes in body weight, ovarian and uterine somatic indices and pattern of estrous cycle in arsenic trioxide exposed rats co-administered with/without HPD.

	Body wt. (g)		Relative wt. (mg/100 g body wt.)
Treatment	Initial	Final	Ovary	Uterus	Number of cycles/rat^a
Control	116 ± 4.0	117 ± 3.6	54.3 ± 4.2	161.7 ± 24.6	5.71 ± 0.18
As2O3^b	114 ± 8.2	115 ± 5.0	35.8 ± 1.6*	94.44 ± 13.2**	4.33 ± 0.21*
As2O3+HPD^c	114 ± 7.8	120 ± 6.2	45.8 ± 1.2	156.8 ± 13.8	5.83 ± 0.17

^aThe number of regular cycles during the course of treatment.

^bCycling rats were treated with As₂O₃ at the dose level of 3 ppm/rat/d for 30 consecutive days and maintained on routine laboratory diet.

^cCycling rats were treated with As₂O₃ at the dose level of 3 ppm/rat/d and maintained on high-protein diet for 30 consecutive days.

Each value represents mean ± SEM (n = 8). The data were analyzed by one-way ANOVA followed by Bonferroni test. All comparisons were made with respect to the respective control values [*p < 0.001; **p < 0.05].

Figure 1. (a) Ovarian section of control rats showing follicles at different stages of maturation and recent ovulation is documented by few functional corpora lutea scattered throughout the periphery (100×). (b) Ovarian section of arsenic-treated rats exhibiting scarce number of follicles and atretic cells with arrested growth at preantral stage (arrow) and a significant part of the stroma is filled by fibrous connective tissue (100×). (c) In greater than 60% of the HPD-supplemented group ovaries, there is major restoration of follicular growth and maturation. Presence of few fully functional corpora lutea marks recent past ovulation (100×).

Figure 2. (a) Uterine section from control rats showing normal histological features (100×). (b) Significant thinning of uterine luminal epithelium, myometrium and endometrium with a consequent reduction in the diameter of the uterine horn in arsenic intoxicated rats (100×). (c) Restoration of normal histological features of the uterus in HPD-supplemented group (100×).

Table 2. Changes in uterine histometry in arsenic trioxide exposed rats co-administered with/without HPD.

Treatment	Diameter (mm)	Epithelial height (µm)	Endometrial thickness (µm)	Myometrial thickness (µm)
Control	2.05 ± 0.02	27.46 ± 1.4	324.0 ± 2.0	322.3 ± 18.0
As2O3^a	1.23 ± 0.04*	10.76 ± 0.6*	302.9 ± 0.8**	149.7 ± 8.6*
As2O3 + HPD^b	1.51 ± 0.03*	25.27 ± 0.6	317.6 ± 4.9	320.2 ± 22.0

^aCycling rats were treated with As₂O₃ at the dose level of 3 ppm/rat/d for 30 consecutive days and maintained on routine laboratory diet.

^bCycling rats were treated with As₂O₃ at the dose level of 3 ppm/rat/d and maintained on high-protein diet for 30 consecutive days.

Each value represents mean ± SEM (n = 8). The data were analyzed by one-way ANOVA followed by Bonferroni test. All comparisons were made with respect to the respective control values [*p < 0.05; **p < 0.001].

Figure 3. (a) Comparative ovarian Δ⁵, 3β-HSD and 17β-HSD activities in the control and arsenic-treated groups with or without HPD supplementation (mean ± SEM), n = 8. (b) Comparative estradiol levels in the control and arsenic-treated groups with or without HPD-supplementation (mean ± SEM), n = 8. *p < 0.05 and **p < 0.001.

Table 3. Effect of HPD co-administration on ovarian MDA level, SOD activity and DNA damage in arsenic trioxide-treated rats.

Treatment	MDA (nM/mg tissue)	SOD (unit/mg tissue)	DNA damage Tail length (µm)
Control	15.10 ± 2.45	2.18 ± 0.19	0.0006 ± 0.00005
As2O3^a	29.51 ± 3.44*	1.33 ± 0.18*	3.8030 ± 0.3040**
As2O3 + HPD^b	14.23 ± 1.17	2.19 ± 0.25	0.0170 ± 0.0100**

^aCycling rats were treated with As₂O₃ at the dose level of 3 ppm/rat/d for 30 consecutive days and maintained on routine laboratory diet.

^bCycling rats were treated with As₂O₃ at the dose level of 3 ppm/rat/d and maintained on high-protein diet for 30 consecutive days.

Each value represents mean ± SEM (n = 8). The data were analyzed by one-way ANOVA followed by Bonferroni test. All comparisons were made with respect to the respective control values [*p < 0.05; **p < 0.001].

Figure 4. DNA fragmentation in ovarian cells as evaluated by single cell gel electrophoresis in control (a), and arsenic-treated without (b) or with (c) HPD supplementation; 100×, (400×, inset).