Dual targeting of TNF-α and free radical toxic stress as a promising strategy to manage experimental polycystic ovary

Figures & data

Table 1. Comparative assessment of histopathological findings of ovary in experimental and control groups and the protective effects of ANP and CLV.

	Control	PCO	ANP	PCO + ANP	CVL	PCO + CVL
Body weight (g)	236.13 ± 3.18	256.38 ± 3.46^a	234.38 ± 4.50	236.50 ± 2.45^b	235.88 ± 3.14	235.13 ± 1.25^b
Ovary weight (g)	0.043 ± 0.004	0.059 ± 0.002^a	0.041 ± 0.001	0.045 ± 0.001^b	0.042 ± 0.002	0.043 ± 0.002^b
Mean no. of atretic primordial follicles	26.82 ± 2.64	48.54 ± 2.84^a	27.27 ± 3.85	34.18 ± 2.89^a,b	25.18 ± 2.96	32.27 ± 3.58^a,b
Mean no. of atretic growing follicles	43.45 ± 4.27	77.73 ± 4.38^a	42.45 ± 6.64	48.82 ± 3.03^b	39.82 ± 4.28	45.27 ± 3.50^b
Mean no. of atretic graafian follicles	6.78 ± 1.19	21.54 ± 2.80^a	5.82 ± 1.33	10.73 ± 1.79^a,b	4.82 ± 1.17	8.73 ± 2.19^b
Mean no. of cystic primordial follicles	0	0	0	0	0	0
Mean no. of cystic growing follicles	0	6.82 ± 1.17^a	0.27 ± 0.47	1.54 ± 0.52^a,b	0	1.36 ± 0.50^a,b
Mean no. of cystic graafian follicles	0.64 ± 0.50	12.36 ± 1.43^a	0.54 ± 0.15	2.18 ± 0.60^a,b	0.45 ± 0.16	1.36 ± 0.50^b
Mean no. of corpus luteum (CL)	4.64 ± 1.03	0.09 ± 0.30^a	4.73 ± 1.00	3.18 ± 0.75^a,b	6.18 ± 1.17^a	4.18 ± 0.75^b
Luteinization of follicular wall and granulosa cells median (min–max)	0 (0–0)	4 (3–4)^a	0 (0–0)	2 (2–2)^b	0 (0–0)	0 (0–1)^b
Vascularization within follicular wall median (min–max)	0 (0–0)	3 (2–4)^a	0 (0–0)	1 (1–2)^b	0 (0–0)	1 (1–2)^b
Oocyte degeneration median (min–max)	0 (0–0)	4 (3–4)^a	0 (0–0)	2 (1–2)^b	0 (0–0)	0 (0–1)^b
Pyknosis of granulosa cells median (min–max)	0 (0–0)	4 (2–4)^a	0 (0–0)	1 (1–2)^b	0 (0–0)	1 (1–2)^b
Granulosa cells Dissociation median (min–max)	0 (0–0)	4 (3–4)^a	0 (0–0)	2 (2–2)^b	0 (0–0)	1 (1–2)^b
Hyperplasia of theca cells median (min–max)	0 (0–0)	4 (3–4)^a	0 (0–0)	1 (1–1)^b	0 (0–0)	0 (0–1)^b

ANP, ANGIPARS™; CVL, carvedilol; PCO, polycystic ovary; min, minimum; max, maximum.

^aRepresents a significant difference between control and other treated groups at p < 0.05 level.

^bRepresents a significant difference between PCO and PCO + ANP and PCO + CVL treated groups at p < 0.05 level.

(0–4): Semi-quantitative microscopic scores in each treated group in terms of ovarian histopathological changes in comparison with the control group (p < 0.05).

Figure 1. Ovarian sections of control and experimental groups. (A) Lower magnification (40×) of ovarian section showing normal folliculogenesis as well as several CL from the control rat. (B) Higher magnification (400×) of control ovary showing fresh CL. (C) Higher magnification (400×) of two intact antral follicles with a clear oocyte surrounded by cumulus cells and several layers of granulosa cells (×400). (D) Section of ovary from PCO rats showing multiple fluid-filled sub-capsular cysts (40×). (E) Higher magnification (400×) of a fluid-filled large cystic follicle of PCO rats with a degenerated thin layer of granulosa cells and hyperplasia of theca cells (×400). (F) Higher magnification (400×) of one antral degenerating atretic follicle showing detached floating oocyte. (G) An antral atretic follicle with a malformed oocyte and luteinized theca cells (400×). (H) Section of the ovary from the PCO + CVL group showing normal developing follicles and several CL (×40). (I) Section of the ovary from the PCO + ANP group showing normal developing follicles and several CL (×40). AF, antral follicle; AAF, atretic antral follicle; CF, cystic follicle; CL, corpora lutea; LGW, luteinized follicular wall; MF, mature follicle; MO, malformed oocyte.

Figure 2. Comparative assessment of serum LPO in control and experimental groups of rats. ^aA significant difference between control and other treated groups at p < 0.05 level. ^bA significant difference among PCO−, PCO + ANP−, and PCO + CVL-treated groups at p < 0.05 level. CVL, carvedilol; ANP, ANGIPARS™; PCO, polycystic ovary; LPO, lipid peroxidation.

Figure 3. Comparative assessment of ovarian LPO in control and experimental groups of rats. ^aA significant difference between control and other treated groups at p < 0.05 level. ^bA significant difference among PCO−, PCO + ANP−, and PCO + CVL-treated groups at p < 0.05 level. CVL, carvedilol; ANP, ANGIPARS™; PCO, polycystic ovary; LPO, lipid peroxidation.

Figure 4. Comparative assessment of ROS in serum in control and experimental groups of rats. ^aA significant difference between control and other treated groups at p < 0.05 level. ^bA significant difference among PCO−, PCO + ANP−, and PCO + CVL-treated groups at p < 0.05 level. CVL, carvedilol; ANP, ANGIPARS™; PCO, polycystic ovary; ROS, reactive oxygen species.

Figure 5. Comparative assessment of ROS in ovary in control and experimental groups of rats. ^aA significant difference between control and other treated groups at p < 0.05 level. ^bA significant difference among PCO−, PCO + ANP−, and PCO + CVL-treated groups at p < 0.05 level. CVL, carvedilol; ANP, ANGIPARS™; PCO, polycystic ovary; ROS, reactive oxygen species.

Figure 6. Comparative assessment of TAP in serum in control and experimental groups of rats. ^aA significant difference between control and other treated groups at p < 0.05 level. ^bA significant difference among PCO−, PCO + ANP−, and PCO + CVL-treated groups at p < 0.05 level. CVL, carvedilol; ANP, ANGIPARS™; PCO, polycystic ovary; TAP, total antioxidant power.

Figure 7. Comparative assessment of TAP in ovary in control and experimental groups of rats. ^aA significant difference between control and other treated groups at p < 0.05 level. ^bA significant difference among PCO−, PCO + ANP−, and PCO + CVL-treated groups at p < 0.05 level. CVL, carvedilol; ANP, ANGIPARS™; PCO, polycystic ovary; TAP, total antioxidant power.

Figure 8. Comparative assessment of serum TNF-α concentration in control and experimental groups of rats. ^aA significant difference between control and other treated groups at p < 0.05 level. ^bA significant difference among PCO−, PCO + ANP−, and PCO + CVL-treated groups at p < 0.05 level. CVL, carvedilol; ANP, ANGIPARS™; PCO, polycystic ovary; TNF-α, tumor necrosis factor alpha.

Figure 9. Comparative assessment of serum insulin concentration in control and experimental groups of rats. ^aA significant difference between control and other treated groups at p < 0.05 level. ^bA significant difference among PCO−, PCO + ANP−, and PCO + CVL-treated groups at p < 0.05 level. CVL, carvedilol; ANP, ANGIPARS™; PCO, polycystic ovary.

Figure 10. Comparative assessment of serum testosterone concentrations in control and experimental groups of rats. ^aA significant difference between control and other treated groups at p < 0.05 level. ^bSignificant difference among PCO−, PCO + ANP−, and PCO + CVL-treated groups at p < 0.05 level. CVL, carvedilol; ANP, ANGIPARS™; PCO, polycystic ovary.

Figure 11. Comparative assessment of serum progesterone concentrations in control and experimental groups of rats. ^aA significant difference between control and other treated groups at p < 0.05 level. ^bA significant difference among PCO−, PCO + ANP−, and PCO + CVL-treated groups at p < 0.05 level. CVL, carvedilol; ANP, ANGIPARS™; PCO, polycystic ovary.

Figure 12. Comparative assessment of serum estradiol concentrations in control and experimental groups of rats. ^aS significant difference between control and other treated groups at p < 0.05 level. ^bA significant difference among PCO−, PCO + ANP−, and PCO + CVL-treated groups at p < 0.05 level. CVL, carvedilol; ANP, ANGIPARS™; PCO, polycystic ovary.