BPA, a diphenyl compound containing groups, that make it structurally similar to synthetic estrogen and is considered as one of the major endocrine disruptors. Silymarin has extensively been used to prevent and/or alleviate some human disease, especially for the treatment of adverse liver conditions. It has an antioxidative efficacy and cancer preventive efficacy. Therefore, we examined the hypothesis that silymarin can inhibit BPA-induced toxicity in boar sperm duing in vitro storage. Sperm characteristics (motility, viability, membrane integrity and mitochondrion activity) in semen exposed to BPA (10~200 uM) were sharply lowered, while it increase in a dose and time dependent manner due to silymarin addition (50~200 uM) into semen extender in the presence of BPA (100 uM). All of the evaluated characteristics were gradually improved in the groups that were treated with silymarin (50~200 uM) in the presence of BPA (100 uM) in comparison to BPA 100 uM alone group, irrespective of incubation periods (3 and 6 h). These results demonstrate that silymarin can ameliorate the toxicity of BPA on boar sperm characteristics during in vitro storage, suggesting that silymarin indirectly act as an antioxidant.

본 연구는 BPA 및 nicotine 첨가 농도와 배양 시간이 돼지 난자의 체외성숙에 미치는 영향을 조사하였다. BPA와 nicotine이 첨가된 TCM-199배양액에서 시간 난자를 배양했을 때 체외성 숙율을 조사하였다. BPA농도가 높을수록 체외성숙율이 유의적으로 낮게 나타났다. BPA를 첨가한 TCM-199 배양액에서 난자를 44시간 배양했을 때 체외성숙율은 각각 로서 첨가 농도가 증가할수록 낮은 체외성숙율을 나타냈다. 난자를 nicotine를 첨가

본 연구는 내분비계 장애물질인 BPA와 DEHP가 돼지 정액 성상(운동성, 생존율, 원형질막의 정상성, 기형율)에 유해한 영향을 미치는지를 검토하였다. 일정 농도로 희석한 돼지 정액에 BPA와 DEHP를 각각 0, 1, 10, 100 μM의 농도로 처리하여 3, 6, 9시간 동안 체외배양을 실시하였다. BPA 처리에 따른 운동성과 생존율은 체외배양 시간과 첨가농도에 따라 감소하였으며, 체외배양시간이 경과함에 따라 대조구와 명백한 차이를 나타냈다. 100 μM의 농도로 처리한 경우의 운동성과 생존율은 체외배양시간에 관계없이 처리군이 대조구에 비해 유의적으로 감소하였다(p<0.05). DEHP 처리에 따른 정자의 운동성과 생존율도 체외배양시간과 첨가농도에 따라 감소하여 BPA의 성적과 유사한 경향을 나타냈다. 정자원형질막의 기능성은 배양시간에 따라 감소하였으며 BPA 및 DEHP 첨가농도 간에 차이가 인정되었으며, 특히 100 μM 처리구에서 급격히 감소하는 성적을 나타났다. 정자의 기형율은 BPA 및 DEHP 처리시간 및 농도에 크게 영향을 받지는 않았다. 본 연구의 결과로 볼 때, 고농도의 BPA와 DEHP(>10 μM)에 정자의 장시간 노출은 유해한 영향을 줄 것으로 사료된다.

Although the toxicological impacts of the xenoestrogen bisphenol-A (BPA) have been studied extensively, but its mechanism of action is poorly understood. Eventually, no standard method exists for evaluating the possible health hazard of BPA. Considering mice spermatozoa as a potential in vitro model, here we demonstrated the effects of BPA exposure (0.0001, 0.01, 1, and 100 μM for 6 h) on spermatozoa and the related mechanisms of action. Our results demonstrated that high concentrations of BPA negatively affect sperm motility, viability, intracellular ATP, and mitochondrial functions by activating the mitogen-activated protein kinase, phosphatidylinositol 3-kinase, and protein kinase-A pathways. The same doses were also employed to identify the differential expressed proteins of exposure and screen their functional affiliation to diseases using sperm proteomics and informatics, respectively. Our results demonstrated that a high concentration of BPA (100 μM) induced differential expression (> 2-fold) of 24 proteins in spermatozoa (16 down- and 9 up-regulated), that are putatively involved in the pathogenesis of several diseases. To the best of our knowledge, this is the first study to demonstrate the mechanisms of BPA action in spermatozoa and to identify the possible biomarkers of exposure. Moreover, we anticipated that current strategy might apply for the hazard assessment of other toxicological agents.

As an endocrine disruptor, bisphenol-A (BPA) causes several functional and behavioral abnormalities related to reproduction. The current study was design to evaluate the effect of perinatal exposure of female mice to BPA on sperm function of adult F(1) offspring. Pregnant female mice F(0) were gavaged with three different concentration of BPA, such as 50 μg/kg/day (tolerable daily intake value by the European Food Safety Authority), 5 mg/kg/day (no-observed-adverse-effect level; NOAEL), and 50 mg/kg/day (lowest-observed-adverse-effect level; LOAEL) and corn oil (7 mg/kg/day; vehicle control). The functional parameters of F(1) spermatozoa were studied both before and after capacitation, whereas the fertility assessment was evaluated by in vitro and in vivo assay using unexposed females. Our results showed that spermatozoa hyperactivated motility, capacitation, intracellular ATP, Ca2+, and ROS levels after capacitation were significantly affected using NOAEL and LOAEL concentration of BPA. However, the sperm motility was only affected by LOAEL dose after capacitation. All of the tested parameters were potentially unaffected by BPA before capacitation, except intracellular ATP that decreased by all concentrations. Although both NOAEL and LOAEL concentration were effectively reduced the rate of fertilization and embryonic development in vitro, however the average litter size was only affected by LOAEL dose. Our finding suggested that perinatal exposure of 50 μg/kg/day did not produce significant effects; however both NOAEL and LOAEL affects overall sperm function after capacitation, leading to impairments in the fertility of F(1) male offspring.

This study was carried out to evaluate the preventive effect of three forms of Korean ginseng roots (fresh, white and red) against bisphenol A (BPA) toxicity in mouse male germ cells (GC-2spd, TM3, TM4). ROS (reactive oxygen species) generation were measured by DCF-DA (2’,7’-dichlorohydrofluorescein diacetate) assay. Also, semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) was performed to quantify the mRNA expression levels of apoptosis- related genes, Bax (pro-apoptotic gene) and Bcl2 (anti-apoptotic gene). ROS generation was increased by 50 μM BPA, but definitely decreased by treatment with Korean ginseng extracts (fresh, white and red) in mouse male germ cells. In especial, Korean fresh ginseng extract reduced significantly ROS production to normal control. In addition, Korean fresh and white ginseng extracts suppressed the apoptosis of mouse male germ cells by fine-tuning mRNA levels of apoptotic genes changed by BPA. In general, Korean fresh ginseng extract was more effective than white ginseng extract for reducing BPAinduced oxidative stress and apoptosis in mouse male germ cells. Therefore, Korean fresh and white ginseng may help to alleviate biphenol A toxicity in mouse male germ cells.

Estrogen is a primary steroid hormone to govern cell fates in the endometrium. It induces expression of a spectrum of genes such as early growth response 1 (Egr1) critical for dynamic change of uterine environments for embryo implantation. Egr1 belongs to the Egr family of zinc finger transcription factors consisting of 4 members (Egr1 to Egr4) that are co-expressed in many different tissues, suggesting that they may have some redundant functions. Bisphenol A (BPA) is a well-known endocrine disruptor with potent estrogenic activity on reproductive system. Here we have demonstrated molecular pathway(s) by which estrogen (17β estradiol, E2) and BPA regulates Egr1 in uterus. Eight-week-old female mice were ovariectomized (OVX) and rested for a week. Uteri of OVX mice treated with E2, BPA and/or progesterone (P4) were collected 2 h after hormone treatment unless otherwise indicated. ICI 182,780 [estrogen receptor (ER) antagonist] and RU486 [progesterone receptor (PR) antagonist] were pretreated 30 min before hormone treatment. Collected uteri were mainly utilized for RT-PCR, realtime-RT-PCR and Western blotting. Egr1 mRNA was rapidly induced with the highest level at 2h after E2 treatment and gradually decreased to basal levels at 12 h. Pretreatment of ICI 182,780 effectively inhibited E2-induced phosphorylation of ERK1/2 and AKT as well as Egr1 transcription. U0126 (a pharmacological ERK1/2 inhibitor), but not Watmannin (a AKT inhibitor), significantly blocked E2-induced Egr1 expression as well as ERK1/2 phosphorylation in the uterus. P4 effectively dampened E2-dependent Egr1 transcription, and its antagonistic effects were partially interfered with RU486 pretreatment. Interestingly, Egr2 and Egr3 showed similar hormone-dependent expression profiles to that of Egr1 in the uterus. BPA (100 mg/kg) was able to induce immediate expression of Egr1 as effective as E2 at 2 h after treatment. ICI 182,780 and P4 considerably reduced BPA-induced expression of Egr1. In addition, RU486 counteracted inhibitory action of P4 on BPA-induced expression of Egr1. While overall patterns of BPA- induced expression of Egr2 and Egr3 were similar to that of Egr1, BPA was not as effective as E2 for induction of Egr2 and Egr3. BPA could induce phosphorylation of ERK1/2 as well as expression of Egr family members, too. Collectively, these results strongly suggest that BPA as well as E2 can activate concurrent expression profiles of Egr family members via ER-ERK1/2 pathways in the uterus.