【摘 要】
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该研究旨在探讨苯并(a)芘[Benzo(a)pyrene,B(a)P]对孕早期小鼠卵巢黄体功能的影响及机制.体内模型:将昆明小鼠每晚按雌雄3∶1的比例合笼,次晨查得阴栓记为孕第1天(d1);将其随
【机 构】
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重庆医科大学公共卫生与管理学院生殖生物学研究室,重庆医科大学生殖与发育国际合作联合实验室,重庆400016
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该研究旨在探讨苯并(a)芘[Benzo(a)pyrene,B(a)P]对孕早期小鼠卵巢黄体功能的影响及机制.体内模型:将昆明小鼠每晚按雌雄3∶1的比例合笼,次晨查得阴栓记为孕第1天(d1);将其随机分为对照组和B(a)P处理组,每日早晨称重后以0.1 mL/10 g动物体质量灌胃给予0.2 mg/(kg·d)的B(a)P,对照组灌胃等体积的玉米油,收取d4、d7小鼠卵巢组织.体外模型:培养小鼠卵巢颗粒细胞KK-1,将其分为对照组(0.1%DMSO)、HCG组(1.0 IU/mL HCG)、HCG+BPDE(1.0 IU/mL HCG和0.5 μmol/L BPDE)联合处理组,处理细胞24 h后进行后续检测.ELISA检测小鼠血清雌激素(E2)、孕激素(P4)水平;qRT-PCR检测体内外卵巢雌、孕激素合成限速酶3β-HSD、17β-HSD和P450SCC的mRNA水平;免疫组化检测卵巢组织切片中Ki67、PCNA的表达,CCK-8检测KK-1细胞增殖情况;Western blot、免疫组化和免疫荧光检测周期相关蛋白CyclinA1、CDK2、CDK4、CyclinB1以及GAS1的表达情况.透射电镜和Mitotracker探针观察线粒体形态.与对照组相比,B(a)P暴露导致孕早期小鼠血清中E2、P4水平明显降低;同时,卵巢雌、孕激素合成限速酶3β-HSD、17β-HSD和P450SCC mRNA水平下调;CCK-8结果显示,BPDE暴露导致细胞活力下降;体内B(a)P暴露导致卵巢黄体中Ki67、PCNA表达下调;Western blot、免疫组化和免疫荧光结果显示,B(a)P或BPDE暴露下调细胞周期相关因子CyclinA1、CDK2、CDK4、CyclinB1及GAS1水平;电镜和免疫荧光结果显示,BPDE暴露导致线粒体形态异常.B(a)P及其代谢物BPDE干扰细胞周期调控,影响线粒体功能,进而导致孕早期小鼠卵巢黄体功能异常.
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