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目的:探讨孕早期Ba P暴露对胚胎着床的影响,为进一步完善Ba P对雌性生殖功能的影响提供实验依据。方法:将60只孕鼠随机分为对照组和Ba P组,每组30只,每组再随机分成3小组,每组10只。从妊娠第1天对照组用玉米油、Ba P组用0.2 mg/(kg·d)剂量Ba P进行灌胃染毒,至妊娠的第4天(D4)、第5天(D5)、第6天(D6)处死并取材。计数小鼠胚胎着床点数目,ELISA方法检测小鼠血浆中雌激素和孕激素水平,荧光定量PCR、Western blot及免疫组织化学方法检测小鼠子宫内膜中雌激素受体-α(estrogen receptor-α,ERα)和孕激素受体(progesterone receptor,PR)水平。结果:比较对照组与Ba P组的胚胎着床数,Ba P组的着床点数在妊娠D5和D6均低于对照组,差异有统计学意义(P<0.05);Ba P组血浆雌激素水平和孕激素水平均高于对照组,差异均有统计学意义(P<0.05),不同染毒时间的雌激素水平与孕激素水平不同,由高到低依次为妊娠D6、D5、D4,各时间点的差异均有统计学意义(P<0.05);ERα的m RNA水平在Ba P组的表达高于对照组,差异有统计学意义(P<0.05),不同染毒时间ERα的m RNA水平不同,妊娠D6的表达高于妊娠D4和D5,差异有统计学意义(P<0.05),PR的m RNA水平在Ba P组的表达低于对照组,差异有统计学意义(P<0.05)。Western blot和免疫组化结果也表明苯并(a)芘暴露组小鼠子宫中ERα表达量升高,PR表达量下降。Western blot和免疫组化结果也表明Ba P组小鼠子宫中ERα表达量升高,PR表达量下降。结论:Ba P可能通过影响孕鼠雌孕激素水平及PR的表达模式,最终导致胚胎着床点数的下降。
OBJECTIVE: To investigate the effect of Ba P exposure during early pregnancy on embryo implantation and to provide experimental evidence for further improving the effect of Ba P on female reproductive function. Methods: Sixty pregnant rats were randomly divided into control group and BaP group, 30 rats in each group. Each group was randomly divided into 3 groups of 10 rats. From the first day of gestation, corn oil was used as the control group, Ba P group was given intragastrically Ba Ba 0.2 mg / (kg · d) to the first day of gestation (D4), the fifth day (D5) 6 days (D6) were sacrificed and drawn. The number of mouse embryo implantation sites was counted. The levels of estrogen and progesterone in plasma of mice were detected by ELISA. The expression of estrogen receptor-α in mouse endometrium was detected by real-time quantitative PCR, Western blot and immunohistochemistry. -α, ERα) and progesterone receptor (PR). RESULTS: Compared with control group and BaP group, the number of implantation sites in BaP group was significantly lower at D5 and D6 than that in control group (P <0.05) (P <0.05). The levels of estrogen and progesterone in different exposure time were different from high to low in order of D6, D5, D4, (P <0.05). The mRNA level of ERα in BaP group was higher than that in control group (P <0.05). The mRNA expression levels of ERα The expression of D6 in pregnancy was higher than that of D4 and D5 in pregnancy (P <0.05), and the expression of m RNA in PR was lower than that in control group (P < 0.05). Western blot and immunohistochemistry also showed that the expression of ERα in the uterus of the mice exposed to benzo (a) pyrene increased, while the expression of PR decreased. The results of Western blot and immunohistochemistry also showed that the expression of ERα increased and the expression of PR decreased in the Ba P group. Conclusion: Ba P may affect the level of estrogen and progesterone in pregnant rats and the expression pattern of PR, leading to the decrease of embryo implantation points.