聚苯乙烯纳米塑料致雄性小鼠睾丸睾酮合成障碍的作用机制研究  

Mechanism of Action of Testosterone Synthesis Disorder in Male Mice Induced by Polystyrene Nanoplastics

作　　者：缪心媛 许晴雨 夏苏干 林天金 蔡国栋[1] 邹辉[1] 顾建红[1] 袁燕[1] 刘宗平[1] 卞建春 MIAO Xinyuan;XU Qingyu;XIA Sugan;LIN Tianjin;CAI Guodong;ZOU Hui;GU Jianhong;YUAN Yan;LIU Zongping;BIAN Jianchun(College of Veterinary Medicine,Yangzhou University,Yangzhou 225009,China;Jiangsu Collaborative Innovation Center for Prevention and Control of Important Epidemics and Zoonoses,Yangzhou 225009,China;Animal Disease Prevention and Control Center of Dantu District,Zhenjiang City,Jiangsu Province,Zhenjiang 212028,China)

机构地区：[1]扬州大学兽医学院,扬州225009 [2]江苏省重要疫病与人兽共患病防控协同创新中心,扬州225009 [3]江苏省镇江市丹徒区动物疫病预防控制中心,镇江212028

出　　处：《中国畜牧兽医》2025年第4期1488-1498,共11页China Animal Husbandry & Veterinary Medicine

基　　金：“十四五”国家重点研发支持项目(2023YFD1801100);高等学校学科创新引智计划资助(D18007);江苏高校优秀学科建设工程资助项目(PAPD);江苏省(扬州大学)研究生科研与实践创新计划项目(SJCX23-2066)。

摘　　要：【目的】探究聚苯乙烯纳米颗粒(NPs)对雄性小鼠生殖系统造成的损伤和影响睾酮合成的作用机制。【方法】将24只清洁级雄性C56小鼠随机分为4组,分别为对照组及低(0.1 mg/L)、中(1 mg/L)、高(10 mg/L)剂量聚苯乙烯NPs组,每组6只,饮水暴露,试验期90 d。试验结束后,小鼠眼球采血,脱颈处死后剥离附睾及睾丸,采集精子。睾丸称重,统计睾丸指数,利用HE染色观察睾丸组织病理变化,使用综合精液分析系统分析精子总活率及精子总活力;利用ELISA试剂盒检测睾丸组织丙二醛(MDA)含量、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)活性以及血清睾酮含量;利用Western blotting检测氧化应激、凋亡相关蛋白表达水平;通过实时荧光定量PCR和Western blotting分别检测睾酮合成相关基因的mRNA和蛋白表达水平。【结果】与对照组相比,中、高剂量聚苯乙烯NPs组小鼠睾丸指数显著或极显著升高(P<0.05;P<0.01),小鼠精子总活力及总活率、血清睾酮含量均显著或极显著降低(P<0.05;P<0.01)。ELISA检测结果显示,与对照组相比,各聚苯乙烯NPs处理组小鼠睾丸组织中MDA含量均极显著升高(P<0.01),T-SOD、CAT和GSH-Px活性显著或极显著降低(P<0.05;P<0.01)。Western blotting检测结果显示,与对照组相比,中、高剂量聚苯乙烯NPs组小鼠睾丸组织中核因子E2相关因子2(Nrf2)蛋白表达量极显著升高(P<0.01),3β-羟类固醇脱氢酶(HSD-3β)蛋白表达量均极显著降低(P<0.01);各聚苯乙烯NPs处理组小鼠睾丸组织血红素加氧酶-1(HO-1)蛋白表达量均极显著降低(P<0.01);高剂量聚苯乙烯NPs组小鼠睾丸组织中天冬氨酸半胱氨酸特异性蛋白酶9(Caspase9)、Caspase3、Bcl2相关X蛋白(Bax)/B淋巴细胞瘤-2(Bcl-2)蛋白表达量均极显著升高(P<0.01),胆固醇侧链裂解酶(CYP11A)蛋白表达量显著降低(P<0.05)。实时荧光定量PCR结果显示,与对照组相比,各聚苯乙烯NPs处【Objective】The aim of this study was to explore the effects of polystyrene nanoparticles(NPs)on reproductive system damage and testosterone synthesis in male mice.【Method】A total of 24 clean grade male C56 mice were randomly divided into 4 groups:Control group and low(0.1 mg/L),medium(1 mg/L)and high(10 mg/L)dose polystyrene NPs groups,with 6 mice in each group.The experiment lasted for 90 d.After the test,blood was collected from eyeballs of mice,epididymis and testicles were removed after neck removal,and sperm was collected.Testis were weighed,testicular index was counted,testicular histopathological changes were observed by HE staining,total sperm motility and total sperm viability were analyzed by comprehensive semen analysis system.The content of malondialdehyde(MDA),the activities of superoxide dismutase(SOD),catalase(CAT)and glutathione peroxidase(GSH-Px)in testicular tissue,and the content of serum testosterone were detected by ELISA kit.Western blotting was used to detect the expression of oxidative stress and apoptosis-related proteins.The mRNA and protein expression of genes related to testosterone synthesis were detected by Real-time quantitative PCR and Western blotting,respectively.【Result】Compared with control group,the testicular index of mice in medium and high dose polystyrene NPs groups was significantly or extremely significantly increased(P<0.05 or P<0.01),the total motility and total viability of sperm and serum testosterone content of mice were significantly or extremely significantly decreased(P<0.05 or P<0.01).The ELISA results showed that compared with control group,the MDA content in testis tissue of mice in polystyrene NPs treated groups was extremely significantly increased(P<0.01),and the activities of SOD,CAT and GSH-Px were significantly decreased or extremely significantly decreased(P<0.05 or P<0.01).Western blotting results showed that compared with control group,the expression of nuclear factor E2 related factor 2(Nrf2)protein in testicular tissue of mice in medium

关 键 词：聚苯乙烯 纳米塑料(NPs) 氧化应激 凋亡 睾酮合成 

分 类 号：S859.8[农业科学—临床兽医学]