Effects of daidzein on rat ovary against ischemia-reperfusion

Abstract

Purpose: Our aim was to investigate protective effects of daidzein treatment on ischemia-reperfusion (I/R) injury-induced ovarian tissue by immunohistochemical techniques.

Methods: Thirty Sprague Dawley female rats were categorized into three groups as sham, I/R group, and I/R+daidzein groups. Bloods were analyzed for malondialdehyde (MDA), glutathione peroxidase (GSH), and myeloperoxidase (MPO), and ovaries were processed for histological tissue protocol.

Results: Both MDA and MPO values were increased in I/R group compared to sham and I/R+daidzein groups. GSH content was increased in I/R+daidzein group compared to I/R groups. In I/R group, theca and follicular cells were degenerated with apoptosis and dilatation and congestion, edema. In I/R+daidzein group, daidzein improved pathologies. In the I/R group, Bax expression was positive with follicular cells, granulosa cells and inflammatory cells. In the I/R+daidzein group, positive Bax reaction was observed in the epithelial, antral, and inflammatory cells. In I/R group, Bcl-2 reaction was in germinative epithelial cells, cells of antral follicle. In the I/R+daidzein group, Bcl-2 expression level was reduced after daidzein treatment.

Conclusions: After the I/R procedure, ovarian cells and follicles were degenerated with apoptosis and inflammation. After daidzein treatment, Bax and Bcl-2 signal were decreased. It was observed that daidzein stopped the apoptotic process.

Figure 1. Biochemical parameters (MDA, GSH and MPO) and histological scores of ovarian tissues in sham, IR and IR+Daidzein groups.. (a) MDA values of sham, I/R and I/R+daidzein groups. (b) GSH, MPO and histological scoring for follicular degeneration, inflammation, hemorrhage, Bcl-2 and Bax expression in sham, I/R and I/R+daidzein groups.

Figure 2. Histological staining of ovarian sections. Hematoxylin and eosin staining. (a) Sham group: smooth-looking germinative epithelium (black arrow) and primordial follicle (red arrow), smooth corpus luteum and stromal area (star). (b) IR group: degenerative changes in theca and granulosa cells in the corpus lutea of the antral follicle (black arrow), loss of structural integrity in the germinal epithelium (blue arrow), dilatation, congestion, and edema in the connective tissue areas (red arrow). (c) IR+daidzein group: the epithelial structure is smooth (black arrow), regular arrangement of granulosa cells in the corpus luteum (orange arrow), decrease in the area of inflammation (star). (d) Bax immunostaining group / sham group: positive Bax expression in some cells under the germinative epithelium (black arrows), negative Bax expression in granulosa cells (asterisk). (e) IR group: positive Bax expression in granulosa cells (asterisk), vessel endothelium (red arrow) and inflammation cells (blue arrowhead). (f) IR+Daidzein group: positive Bax reaction in germinative epithelium (black arrow), some cells in the stromal area (orange arrow), antral follicle cumulus oophorus (red arrow). (g) Bcl-2 immunostaining group / sham group: negative Bcl-2 expression in cells in preantral and antral follicle (black arrow), granulosa and theca cells in corpus luteum (star), and Bcl-2 reaction in base macrophage cells in stromal region (red arrow) moderate positivity. (h) IR group: Bcl-2 reaction positive in germinative epithelium (black arrow), antral follicle cumulus cells (red arrow), stromal area (star). (i) IR+daidzein group: positive Bcl-2 expression in germinal epithelial cells (black arrow), inflammatory cells (star) and negative Bcl-2 expression in granulosa cells of follicles (red arrow).