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Frontiers of Medicine

ISSN 2095-0217

ISSN 2095-0225(Online)

CN 11-5983/R

Postal Subscription Code 80-967

2018 Impact Factor: 1.847

Front Med Chin    2009, Vol. 3 Issue (2) : 220-226     DOI: 10.1007/s11684-009-0037-2
Effect of repeated gonadotropin stimulation on ovarian reserves and proliferation of ovarian surface epithelium in mice
Linlin LIANG, Bei XU, Guijin ZHU()
Reproductive Medicine Center, Tongji Hospital, Tongji Medicine College, Huazhong University of Science and Technology, Wuhan 430030, China
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This study aimed to evaluate the effect of repeated ovarian stimulation (OS) on the ovarian follicular population and morphology in female mice and its influence on the embryo’s developmental ability, and the profile of the ovarian surface epithelium (OSE). A total of 75 mice were enrolled in this experiment and randomly assigned into three groups: repeated ovarian stimulated group [n=25; receiving 5 IU pregnant mare serum gonadotrophin (PMSG) and human chorionic gonadotropin (hCG) at 6 day intervals for 5 cycles]; single ovarian stimulated group (n=25; receiving 5 IU PMSG and hCG for 1 cycle), and control group (n=25; without additional treatment). The follicle number at various stages and the morphologies were recorded respectively in the three groups. The harvested oocytes or embryos, cleavage rate, good quality embryo rate, and blastocyst production rate were counted and calculated, and the proliferations of ovarian surface epithelium were evaluated respectively. In the three groups, the single ovarian stimulation treatment significantly increased the mean number of ovarian oocytes or embryos (39.25±10.77 one-cell embryos/female); on the other hand, repeated gonadotropin stimulation obtained the lowest mean number (5.15± 2.81 eggs/female, P<0.01). Repeated ovarian stimulation also tended to decrease normal follicles of primary follicles (66.67%) and secondary follicles (72.86%), and got the lowest cleavage rate (67.47%), lowest good quality embryo rate (2.41%), and lowest blastocyst production rate (0). The OSE cells adjacent to the antral follicles and corpus luteum (CL) in the repeated ovarian stimulated group (81.8%) had a significantly higher proliferation rate than the other groups. The proliferation rate of the OSE in the single ovarian stimulated group (56.4%) was significantly higher than that in the control group (37.5%) (P<0.01). In conclusion, single ovarian stimulation may produce more oocytes/embryos. However, repeated gonadotropin stimulation may have a negative effect on the ovarian follicular quality, the number of mature retrieved oocytes, and the embryo quality, even increasing the chance of ovarian cancer.

Keywords gonadotropin-releasing hormone      ovarian reserve      embryo developmental ability      ovarian surface epithelium     
Corresponding Authors: ZHU Guijin,   
Issue Date: 05 June 2009
URL:     OR
Fig.1  Histological pictures of mouse ovarian tissues of three groups (HE, ×400). (a) Ovarian tissue in the control group, the morphologically normal primary (P) and secondary follicles (S) consisted of intact oocytes and compact granulosa cells. (b) Ovarian tissue subjected to single ovarian stimulated group, the normal secondary follicle (S.N) and the abnormal secondary follicle (S. Ab) are seen. The abnormal secondary follicle (arrow) have a shrunken oocyte. (c) Ovarian tissue subjected to repeated ovarian stimulated group, primary follicles (P) and secondary follicles (S) are morphologically abnormal. The follicles haved shrunken oocytes with pyknotic nuclei.
Fig.2  The percent age of primordial(1), primary(20 and secondary(3) follicles in mouse ovarian tissue in control group, single ovarian stimulated group and repeated ovarian stimulated group
Fig.3  Percentages of morphologically normal follicles in the stages of primary(1) and secondary(2) in control group, single ovarian stimulated group and repeated ovarian stimulated group. : <0.05, compared with control group and single ovarian stimulated group.
Fig.4  Cleavage rate(1), good quality embryo rate(20 and blastocyst rate(3) of control group, single ovarian stimulated group and repeated ovarian stimulated group. Percentage data were shown as ±.: <0.05, compared with control group and single ovarian stimulated group.
Fig.5  The OSE cell counts adjacent to antral follicles and corpus luteum (CL) in three groups. (a) Control group; (b) single ovarian stimulated group; (c) repeated ovarian stimulated group. Data represent the least-square mean (%) of proliferation. A significant difference was shown among groups (<0.05). BrdU in the OSE of cells surrounding antral follicles and CL stained with DAB and counterstained with hematoxylin from postovulatory treatment groups. BrdU-positive cells were indicated by black arrows, BrdU-negative cells were labeled with arrowheads, and proliferating granulosa cells within antral follicles were labeled with red arrows.
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