褪黑素对二苯酮-3暴露小鼠卵母细胞成熟水平及线粒体动力学的影响  

The effect of melatonin on the maturation level of oocytes and mitochondrial dynamics in mice exposed to benzophenone⁃3

作　　者：史若锦 熊钰莹 张雪玲 金龙 朱海瑛 SHI Ruojin;XIONG Yuying;ZHANG Xueling;JIN Long;ZHU Haiying(Guangdong Provin-cial Key Laboratory of Major Obstetric Diseases,Guangdong Provincial Clinical Research Center for Obstetrics and Gynecology,Guangdong-Hong Kong-Macao Greater Bay Area Higher Education Joint Laboratory of Maternal-Fetal Medicine,Department of Obstetrics and Gynecology,the Third Affiliated Hospital of Guangzhou Medical University,Guangzhou 510150,Guangdong,China)

机构地区：[1]广东省产科重大疾病重点实验室,广东省妇产疾病临床医学研究中心,粤港澳母胎医学高校联合实验室,广州医科大学附属第三医院妇产科,广东广州510150 [2]广东省妇幼保健院生殖健康与不孕症科,广东广州511400

出　　处：《实用医学杂志》2024年第23期3275-3283,共9页The Journal of Practical Medicine

基　　金：国家自然科学基金青年科学基金项目(编号:82101733);广州市科技计划项目(编号:2023A04J0543)。

摘　　要：目的探讨褪黑素(melatonin,MT)改善二苯酮-3(benzophenone-3,BP-3)暴露小鼠卵母细胞质量的作用及机制。方法试验将6~12周龄的ICR雌鼠GV期卵母细胞分别在M16培养液、0.8μmol/L BP-3培养液以及1×10-7mol/L MT+0.8μmol/L BP-3混合培养液中进行体外培养。将ICR雌鼠随机分为3组:对照组、BP-3组、BMT组;对照组灌胃0.5 m L纯净水、BP-3组灌胃0.5 m L含0.8μmol/L BP-3的药物溶液、BMT组灌胃0.5 m L含0.8μmol/L BP-3以及15 mg/kg MT的药物溶液,每天1次,连续灌胃4周,进行体内实验。随后通过检测3组小鼠的MⅡ期卵子成熟率,线粒体动力学相关基因Mfn1、Opa1、Fis1、Drp1的转录水平及其蛋白表达水平,线粒体膜电位及纺锤体形态来探讨MT对BP-3暴露小鼠卵母细胞线粒体的挽救作用。结果与对照组相比,MT处理显著增加了BP-3组卵母细胞中线粒体融合基因Mfn1和Opa1的转录及蛋白水平表达,同时下调了线粒体分裂基因Fis1和Drp1的m RNA与蛋白表达。此外,BMT组的卵母细胞ROS水平和纺锤体形态异常比例明显低于BP-3组,但其线粒体膜电位显著升高。结论生理浓度BP-3暴露具有生殖毒性,但添加适当浓度的MT可以明显改善BP-3暴露小鼠卵母细胞的线粒体动力学及其发育潜能。Objective To investigate the effects and mechanisms of melatonin(MT)on improving oocyte quality in mice exposed to benzophenone-3(BP-3).Methods In this study,6~12-week-old female ICR mice were cultured in vitro in M16 culture,0.8μmol/L BP-3 medium and 1×10−7 mol/L MT+0.8μmol/L BP-3 mixed culture.Female ICR mice were randomly segregated into three groups:control,BP-3,and BMT.The control group received 0.5 mL of purified water,the BP-3 group was administered 0.5 mL of a 0.8μmol/L BP-3 solution,and the BMT group received 0.5 ml of a combination of 0.8μmol/L BP-3 and 15 mg/kg MT solution via gavage once daily for four weeks,to facilitate in vivo experimentation.Subsequently,the oocyte maturity rate,transcription levels and protein expression levels of mitochondrial dynamics-related genes Mfn1,Opa1,Fis1 and Drp1,mitochondrial membrane potential and spindle morphology were detected in the three groups to explore the rescue effect of MT on the mitochondria of BP-3-exposed mice.Results Compared to the control group,MT treatment markedly enhanced the transcription and protein levels of the mitochondrial fusion genes Mfn1 and Opa1 in oocytes,while concurrently down-regulating the mRNA and protein levels of the mitochondrial fission genes Fis1 and Drp1.Additionally,the BMT group exhibited significantly lower levels of ROS and abnormal spindle morphology in their oocytes compared to the BP-3 group,yet their mitochondrial membrane potential was notably elevated.Conclusion Physiological concentration of BP-3 exposure was toxic for reproduction,but the addition of appropriate concentrations of MT could significantly improve the mitochondrial dynamics and developmental potential of oocytes in BP-3-exposed mice.

关 键 词：褪黑素 二苯酮-3 线粒体动力学 卵母细胞成熟 

分 类 号：R394[医药卫生—医学遗传学]