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目的:苯并(a)芘(benzo(a)pyrene,Ba P)被证实是对生殖有不良影响的雌激素类似物,研究发现苯并(a)芘影响胚胎着床,本研究检测Ba P对早孕小鼠子宫中Wnt信号通路的影响,以探讨其干扰胚胎着床的机制。方法:将成年雌雄昆明鼠进行交配,次日见阴栓记为妊娠第1天(D1)。将孕鼠随机分为玉米油对照组(n=45)和Ba P处理组(n=45),玉米油对照组从妊娠第1天开始给予玉米油灌胃,Ba P处理组则给予0.2 mg/(kg·d)的Ba P进行灌胃,直至妊娠的第4、5、6天(D4、D5、D6)分别处死,收取子宫材料,计数着床点数目,采用荧光定量PCR,免疫组化法,Western blot分别检测所收集子宫内膜材料中Wnt信号通路相关基因表达情况。结果:苯并(a)芘染毒后,引起胚胎着床数下降,差异具有统计学意义(P=0.000);荧光定量PCR显示Wnt7a m RNA水平在Ba P组较对照组表达下降,差异具有统计学意义(P=0.000);而β-catenin m RNA在Ba P组水平升高,差异具有统计学意义(P=0.002、P=0.003、P=0.011);Western blot和免疫组化也均显示Ba P染毒后Wnt7a的表达下降,β-catenin的表达有所上升。结论:苯并(a)芘可能通过干扰Wnt7a、β-catenin的表达影响小鼠胚胎着床。
OBJECTIVE: Benzo (a) pyrene (BaP) has been shown to be an estrogen analogue that has adverse effects on reproduction. It has been found that benzo (a) pyrene affects embryo implantation. On the Wnt signaling pathway in the uterus of early pregnant mice to explore its mechanism of interfering with embryo implantation. Methods: Male and female Kunming mice mating, the next day to see the pinning of the first day of pregnancy (D1). The pregnant rats were randomly divided into corn oil control group (n = 45) and Ba P treatment group (n = 45). The corn oil control group was orally administered with corn oil from the first day of pregnancy, while the Ba P treatment group received 0.2 mg (kg · d) Ba P was intragastrically administered until pregnancy 4,5,6 days (D4, D5, D6) were sacrificed, the uterus material was collected, counting the number of implantation sites, using quantitative PCR, immune group Western blot were used to detect the expression of Wnt signaling pathway related genes in the endometrium collected. Results: Benzo (a) pyrene exposure decreased the number of embryo implantation, the difference was statistically significant (P = 0.000). Fluorescent quantitative PCR showed that the expression of Wnt7a m RNA in BaP group was lower than that in control group (P = 0.002, P = 0.003, P = 0.011). Western blot and immunohistochemistry also showed that β-catenin m RNA in Ba group was significantly higher than that in Ba group The results showed that the expression of Wnt7a decreased and the expression of β-catenin increased after Ba P exposure. CONCLUSION: Benzo (a) pyrene may affect mouse embryo implantation by interfering with the expression of Wnt7a and β-catenin.