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. 2009 Jun;83(6):601-8.
doi: 10.1007/s00204-008-0400-0. Epub 2009 Jan 23.

Lactational coumestrol exposure increases ovarian apoptosis in adult rats

Hyun-Ju Moon  1 , Ji Hyun Seok, Soon Sun Kim, Gyu Seek Rhee, Rhee Da Lee, Jun Young Yang, Soo Yeong Chae, Seung Hee Kim, Ji Young Kim, Jin-Yong Chung, Jong-Min Kim, Soo Youn Chung
Affiliations

Lactational coumestrol exposure increases ovarian apoptosis in adult rats

Hyun-Ju Moon et al. Arch Toxicol. 2009 Jun.

Abstract

This study is the first to examine the increased apoptosis in the adult rat ovary after lactational exposure to coumestrol (COU), a potent phytoestrogen. Lactating dams were gavaged at doses of 0.01, 0.1, 1, and 10 mg/kg COU during the lactation period and the reproductive effects of female pups were investigated in young adults. Rats were sacrificed at postnatal days (PND) 81-84. Ovarian weights were reduced significantly at 0.1 and 1.0 mg/kg COU. The reduction in the ovarian weight occurred in parallel with an increase in the apoptosis at PND 135-140. A marked dose-dependent increase in the expressions of active caspase-3 and -7 was observed in ovarian granulosa cells. Immunostaining for active caspase-3 and the TUNEL staining of apoptotic cells were also increased in ovaries exposed to COU in a dose-dependent manner. These results suggest new sights into the effect of lactational exposure to COU on the female reproductive health.

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Figures

Fig. 1
Fig. 1
Western blot for active caspase-3 and -7 during cell death (apoptosis) in ovaries of adult rats lactationally exposed to coumestrol (0.01, 0.1, 1, and 10 mg/kg). Arrows indicate the positions of active caspase-3 (17 and 19 kDa) and -7 (19 kDa). These data are representative of results obtained from the analysis of three independent experiments (four rats per group)
Fig. 2
Fig. 2
Immunohistochemistry (IHC) of cleaved caspase-3 in ovaries of adult rats lactationally exposed to coumestrol (0.1, 1, and 10 mg/kg). Magnification, ×250. OC oocyte, GC granulosa cell, TC theca cell, LC luteal cell, AT antrum. These data are representative of results obtained from the immunostaining in ovarian sections from four different rats
Fig. 3
Fig. 3
a In situ detection of apoptotic cells (TUNEL) staining in ovaries of adult rats lactationally exposed to coumestrol (0.1, 1, and 10 mg/kg) during the lactation period. These photographs are representative of results obtained in ovarian sections from four different rats. Arrows point to the apoptotic cells. Magnification, ×250. OC oocyte, GC granulosa cell, TC theca cell, LC luteal cell, AT antrum. b Semi-quantitative evaluation of TUNEL assay. The number of TUNEL-positive follicles was counted in ovarian sections from four animals. Data are mean ± SEM from 4 animals. *P < 0.05 and **P < 0.01 by Duncan’s test
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