孕期纳米银暴露对子代雌性成年小鼠乳腺干细胞分化的影响  

Effect of silver nanoparticles exposure during pregnancy on mammary stem cells differentiation in offspring female adult mice

作　　者：江浩 赵凡 喻家键 王红珠 孙晓伟 高慧 JIANG Hao;ZHAO Fan;YU Jia-jian;WANG Hong-zhu;SUN Xiao-wei;GAO Hui(School of Public Health and Management,Wenzhou Medical University,Wenzhou 325035,China)

机构地区：[1]温州医科大学公共卫生与管理学院,浙江温州325035

出　　处：《中国药理学与毒理学杂志》2020年第5期359-367,共9页Chinese Journal of Pharmacology and Toxicology

基　　金：国家自然科学基金(81801428);温州市科技局项目(Y20180199)。

摘　　要：目的探讨孕期纳米银颗粒(Ag-NP)暴露对子代雌性成年小鼠乳腺干细胞数量和功能的影响,并研究其增加乳腺癌风险的潜在机制。方法孕小鼠于孕第12天分别ig给予超纯水(正常对照组)或Ag-NP 5 mg·kg^-1,7 d后停止暴露。取各组3月龄子代雌性小鼠,测量子宫长度和直径和体质量;洋红-硫酸铝钾染色观察乳腺导管形态,测量淋巴结面积;HE染色观察乳腺导管增生并定量增生比例;流式细胞术检测乳腺干细胞亚群比例;集落形成和免疫荧光实验检测管腔干细胞亚群集落百分比;免疫组织化学方法检测子代小鼠乳腺导管环氧化酶2(Cox-2)和信号转导和转录活化蛋白5(Stat5)蛋白表达水平。结果与正常对照组相比,孕期暴露Ag-NP后子代小鼠的子宫长度和直径无显著影响,但体质量显著升高〔(25.7±2.5)g vs(18.3±2.7)g,P<0.01〕。洋红-硫酸铝钾染色发现,孕期暴露Ag-NP后子代成年雌性小鼠乳腺导管变细,结构发生紊乱。HE染色结果显示,与正常对照组相比,Ag-NP暴露组乳腺导管增生比例增加,为(54.6±2.8)%(P<0.05)。流式结果显示,与正常对照组相比,管腔干细胞数目减少,为(1.1±0.2)%(P<0.01)。集落形成和免疫荧光实验结果显示,与正常对照组相比,Ag-NP暴露组总集落数无显著差异,双阳性(K8+K14+)管腔干细胞亚群集落比例增加,为(78.2±2.2)%(P<0.05)。免疫组化结果显示,与正常对照组相比,Ag-NP暴露组Cox-2和Stat5蛋白表达水平显著升高(P<0.05,P<0.01)。结论孕期Ag-NP暴露导致乳腺干细胞分化的改变,引起干细胞数量的变化,从而增加乳腺癌的发生风险,而该影响可能与Cox-2/Stat5信号通路的激活有关。OBJECTIVE To investigate the effect of silver nanoparticles(Ag-NPs)exposure during pregnancy on the number and function of mammary gland stem cells in offspring female mice,and to explore its potential mechanism of increasing breast cancer risk.METHODS Pregnant mice were given intragastric(ig)ubtrapure water(normal control)or Ag-NPs 5 mg·kg^-1 on the 12th day of pregnancy.The exposure was stopped after 7 d,and the uterine length and diameter of 3-month-old female mice were measured.The mammary duct morphology was observed and the lymph node area was measured with magenta aluminum potassium sulfate staining.HE staining was used to observe and quantify the proportion of mammary duct hyperplasia.Flow cytometry was used to detect the proportion of breast stem cell subsets,while colony formation and immunofluorescence experiments were used to detect the properties of lumen stem cells.Immunohistochemical methods were employed to detect the expres⁃sion levels of cyclooxygenase-2(Cox-2)and signal transducer and activator of transcription 5(Stat5)in the mammary ducts of offspring mice.RESULTS Compared with the normal control group,the uterine length and diameter of offspring mice exposed to Ag-NPs during pregnancy were not significantly affected,but the body mass was significantly increased[(25.7±2.5)g vs(18.3±2.7)g,P<0.01].Magenta aluminum potassium sulfate staining revealed that the mammary ducts of the offspring of adult female mice after exposure to Ag-NPs during pregnancy became thinner and the structure was disorderly.HE staining results showed that compared with the normal control group,the proportion of breast duct hyperplasia in the Ag-NPs exposed group increased,which was(54.6±2.8)%(P<0.05).Flow cytometry results showed that compared with the normal control group,the number of luminal stem cells in the Ag-NPsexposed group decreased by(1.1±0.2)%(P<0.01).The results of colony formation and immunofluores⁃cence showed that there was no significant difference between the normal control and Ag-NPs group in to

关 键 词：纳米银 乳腺干细胞 管腔干细胞 乳腺癌 

分 类 号：R394.6[医药卫生—医学遗传学]