新生期双酚A染毒对小鼠睾丸Boule基因表达的长期影响

Long-term effects of neonatal exposure to bisphenol A on testes structure and the expression of Boule in testes of male mice

目的探讨新生小鼠双酚A染毒对小鼠睾丸结构及Boule基因表达的长期影响及机制。方法将12只清洁级妊娠ICR母鼠,待分娩后随机分为空白对照组、阴性对照组(玉米油)和双酚A(100μg/kg)组。仔鼠从出生第1天开始按4μL/g BW颈背部皮下注射药物,直到出生第21天停止注射,出生第35和70天处死取材。苏木精-伊红染色观察小鼠睾丸组织形态学变化,采用RT-PCR法测定睾丸Boule mRNA表达量变化,免疫荧光和Western blot法检测睾丸内Boule蛋白的表达水平。结果与空白对照组、阴性对照组相比,出生第35和70天的双酚A组睾丸曲细精管直径与生精上皮厚度显著减小,管腔显著增大(P<0.05),睾丸中Boule mRNA和蛋白表达水平均显著降低(P<0.05,P<0.01)。结论新生期双酚A染毒可通过降低睾丸组织内Boule mRNA和蛋白质的表达水平进而影响睾丸发育。

Objective To investigate the long-term effects and the mechanism of neonatal bisphenol A（BPA） exposure on mouse testicular structure and Boule expression. Methods A total of 12 pregnant ICR mice were randomly divided into three groups： blank control group, negative control group （corn oil） and BPA 100 μg/kg group. After delivery, BPA was given daily by neck subcutaneous injection to the offspring male mice from postnatal day （PND 1 ） to PND 21. The offspring male mice were sacrificed on PND 35 and PND 70. Morphological changes of testes were detected with hematoxylin-eosin staining, the level of Boule mRNA expression was determined by RT-PCR, the expression of Boule protein was detected by immunofluorescenee and Western blotting. Results Compared with blank control group and negative control group, the diameter and the epithelium thickness of seminiferous epithelium in the same period （PND 35, PND 70） were significantly decreased and the lumen was significantly increased （P 〈 0.05 ） in the testes of BPA （100 μg/kg） group. In addition, the expressions of Boule mRNA and protein were decreased remarkably （P 〈 0. 05, P 〈 0. 01 ） in testes of BPA 100 μg/kg group. Conclusion Neonatal BPA exposure has a long-term effect on mouse testicular development and may affect testicular development by decreasing the expression of Boule mRNA and protein in testes.