黄河包头段水源水有机提取物对雄性小鼠生表殖达细的胞影凋响亡及睾丸组织3-NT/Caspase-3表达的影响  

Effects of organic extracts in source water in Baotou section of the Yellow River on germ cell apoptosis and 3-NT/Caspase-3 expressions in testis of male mice

作　　者：高艳荣[1] 史雪敏 贾玉巧 王丽[1] 邰代鹏 孙瑶 常翔宇 张晗 田江 刘璐敏 李东颖 GAO Yan-rong;SHI Xue-min;JIA Yu-qiao;WANG Li;TAI Dai-peng;SUN Yao;CHANG Xiang-yu;ZHANG Han;TIAN Jiang;LIU Lumin;LI Dong-ying(School of Public Health,Baitou Medical College,Inner Mongolia University of Science and Technology,Baotou,Inner Mongolia 014040,China)

机构地区：[1]内蒙古科技大学包头医学院公共卫生学院

出　　处：《环境与职业医学》2019年第12期1137-1142,共6页Journal of Environmental and Occupational Medicine

基　　金：包头医学院科学研究基金项目(BYJJ-YF 201712);内蒙古自治区高等学校科学研究项目(NJZCI14262);2019年国家级大学生创新创业训练计划项目(201910127003)

摘　　要：[背景]水体有机物污染对生殖健康的危害已成为目前重点关注的问题。我国水环境污染主要是河流有机物污染。[目的]探讨黄河包头段水源水有机提取物对小鼠生殖细胞凋亡及睾丸组织3-硝基酪氨酸(3-nitrotyrosine,3-NT)和天冬氨酸半胱氨酸特异性蛋白3(cysteinyl aspartate specific proteinase-3,Caspase-3)表达的影响,并初步探讨其生殖毒性作用机制。[方法]60只SPF级雄性ICR小鼠随机分为6组:对照组、黄河水有机提取物染毒组(YRW-OE组)、黄河水有机提取物+N-乙酰半胱酸(NAC)处理组(YRW-OE+NAC组)、自来水有机提取物染毒组(TW-OE组)、自来水有机提取物+NAC处理组(TW-OE+NAC组)、NAC组,每组10只。经口灌胃染毒,隔日1次,每次染毒前1 hYRW-OE+NAC组、TW-OE+NAC组及NAC组小鼠经腹腔以0.02 mL/g(以体重计)体积注射浓度为10 g/L的NAC。染毒4周后,取小鼠附睾及睾丸组织,透射电镜检测生殖细胞凋亡,采用细胞计数板计数精子数量和观察精子畸形,ELISA法检测睾丸组织3-NT的表达,Western blotting法检测睾丸组织Caspase-3表达。[结果]与对照组[(4.15±0.06)×10^7/g,(4.84±0.65)%]比较,YRW-OE组小鼠精子数量[(2.86±0.06)×10^7/g]减少,精子畸形率[(19.59±1.49)%]增加(P<0.05),TW-OE组仅精子畸形率[(7.89±1.16)%]增加(P<0.05);YRW-OE+NAC组小鼠精子数量[(3.55±0.06)×10^7/g]高于YRW-OE组(P<0.05),精子畸形率[(11.44±1.22)%]低于YRW-OE组(P<0.05);TWOE+NAC组小鼠精子畸形率[(5.62±0.91)%]低于TW-OE组(P<0.05)。透射电镜显示,YRWOE组细胞核碎裂,核糖体和溶酶体等细胞器减少,细胞器结构不完整,凋亡细胞多见;YRW-OE+NAC组偶见核染色质固缩,核糖体、内质网和溶酶体等细胞器未见减少。与对照组比较,各染毒组小鼠睾丸组织3-NT和Caspase-3蛋白表达量明显增加,YRW-OE+NAC组3-NT[(8.46±0.40)mg/L]和Caspase-3蛋白(0.38±0.02)表达均低于YRW-OE组[(12.28±0.78)mg/L、0.48±0.02,均P<0.05];TW-OE+NAC组仅3-NT表达低于[Background]Reproductive health hazards sourced from organic pollutants in river water have become a major concern at present.The pollution of river water in China is mainly caused by organic pollutants.[Objective]This experiment investigates the effects of organic pollutants in source water inBaotou section of the Yellow River on the apoptosis of germ cells and the expressions of 3-nitrotyrosine(3-NT)and cysteinyl aspartate specific proteinase-3(Caspase-3)in testis of male mice,and preliminarily explores the relevant mechanism of action.[Methods]Sixty SPF ICR male mice were randomly divided into six groups:control group,Yellow River water organic extracts treatment group(YRE-OE group),Yellow River water organic extracts+N-acetylcysteine(NAC)treatment group(YRE-OE+NAC group),tap water organic extracts treatment group(TW-OE group),tap water organic extracts+NAC treatment group(TW-OE+NAC group),and NAC group,with 10 mice in each group.The YRE-OE were administered to all mice by gavage once every other day,and the mice in the YRE-OE+NAC group,TW-OE+NAC group,and NAC group were intraperitoneally injected with 10 g/L NAC in a volume of 0.02 mL/g(in body weight)1 h before the YRE-OE administration.After four weeks of the exposure,epididymis and testis tissues were isolated to observe germ cell apoptosis under transmission electron microscope;germ cells were counted and sperm malformation was observed;3-NT expression in testis was detected by ELISA;and Caspase-3 protein expression was detected by Western blotting.[Results]Compared with the control group[(4.15±0.06)×10^7/g,(4.84±0.65)%],the YRE-OE group showed decreased germ cells[(2.86±0.06)×10^7/g]and increased sperm malformation rate[(19.59±1.49)%](P<0.05),and the TW-OE group only showed increased sperm malformation rate[(7.89±1.16)%](P<0.05);compared with the YRE-OE group,the YRE-OE+NAC group showed increased germ cells[(3.55±0.06)×10^7/g]and decreased sperm malformation rate[(11.44±1.22)%](P<0.05);compared with the TW-OE group,the TWOE+NAC group showed dec

关 键 词：黄河 有机提取物 睾丸 细胞凋亡 3-NT Caspase-3 

分 类 号：R114[医药卫生—卫生毒理学] R123.7[医药卫生—公共卫生与预防医学]