Histomorphometric and immunohistochemical changes in interstitial cells and ovarian follicles of rats with polycystic ovaries treated with clomiphene citrate

Abstract

Objective

To evaluate the histomorphometric and immunohistochemical changes in interstitial cells and ovarian follicles of rats treated with clomiphene citrate during and after induction of permanent estrus.

Methods

Twenty four adult-female rats with regular estrous cycle were equally divided into three groups: (1) GCtrl–at estrous phase. (2) GPCOS–at permanent-estrous phase. (3) GCC–PCOS rats, which remained exposed to 60 days of continuous illumination and treated with Clomiphene Citrate. After that, the animals were euthanized, and the ovaries were removed and processed for paraffin embedding. Sections were stained with H.E. for histomorphometry or subjected to immunohistochemistry for Ki-67 and cleaved caspase-3 detections.

Results

The GPCOS showed lack of corpus luteum and several ovarian cysts, as well as interstitial-like cells. The presence of corpus luteum and a significant increase in primary and antral follicles were observed in GCC, which also showed a decrease in the number of ovarian cysts and in the area occupied by interstitial-like cells, as well as a decrease in nuclear volume of interstitial cells. The percentage of cell proliferation was significantly higher in granulosa cells of the GCC. On the other hand, the percentage of apoptosis was significantly higher in the granulosa cells of GPCOS than the GCC.

Conclusion

The ovaries of rats treated with clomiphene citrate showed a decrease in the number of cysts, an increase in the number of ovarian follicles, the presence of corpus luteum along with a decrease in the nuclear volume in the area occupied by interstitial cells.

摘要

目的:观察枸橼酸克罗米芬诱导永久发情期前后大鼠间质细胞和卵泡的组织形态和免疫组化变化。

方法:将24只发情周期规律的成年雌性大鼠随机分为3组:(1)GCtrl-发情期。(2) GPCos -永久发情期。(3) GCC-PCOS大鼠, 持续光照60 d, 并给予枸橼酸克罗米芬。之后, 这些动物被安乐死, 摘除卵巢并进行石蜡包埋。切片用H.E.染色进行组织形态测定, 进行免疫组化检测Ki-67和cleaved caspase-3。

结果:GPCOS组表现为缺乏黄体、多个卵巢囊肿及间质样细胞。GCC中可见黄体存在, 原始卵泡和窦卵泡明显增多, 卵巢囊肿数量减少, 间质样细胞区域减少, 间质细胞核体积缩小。GCC组颗粒细胞的增殖率明显高于对照组。另一方面, GPCOS颗粒细胞的凋亡率明显高于GCC。

结论:枸橼酸克罗米芬组大鼠卵巢囊肿减少, 卵泡增多, 黄体存在, 间质细胞区域缩小。

Keywords:

• Ovary
• Polycystic ovary syndrome
• clomiphene citrate

Disclosure statement

No potential conflict of interest was reported by the author(s).