摘要
目的探讨乙烯利暴露对青春期雄性SD大鼠精子质量的影响及机制。方法选取45日龄雄性SD大鼠40只,按照随机数字表法分为对照组和低、中、高剂量实验组,每组10只,分别给予9 g/L盐水溶液和100 mg/kg、200 mg/kg、400 mg/kg的乙烯利水溶液连续灌胃28 d。取一侧附睾尾制备精子混悬液,检测精子浓度及活力;取睾丸、附睾组织行HE染色,光镜下观察睾丸、附睾组织病理形态变化;检测附睾超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)活性,丙二醛(MDA)水平;采用酶联免疫吸附测定(ELISA)试剂盒检测附睾α-葡萄糖苷酶活性、左旋肉碱(LC)水平、核转录因子E-2相关因子(Nrf2)及肉碱转运蛋白(OCTN2)表达水平,以评估乙烯利对附睾的氧化损伤。采用SPSS 26.0软件进行分析,多组间比较采用单因素方差分析,2组间均数比较采用LSD法。结果对照组、低、中、高剂量组精子浓度分别为(40.21±1.94)×10^(9)/L、(35.23±2.53)×10^(9)/L、(23.61±2.62)×10^(9)/L、(18.86±2.16)×10^(9)/L;活动率分别为(70.98±3.01)%、(57.96±3.75)%、(45.71±2.41)%、(31.23±2.26)%;实验组大鼠附睾精子浓度及活力均明显低于对照组(均P<0.01);对照组,低、中、高剂量组SOD活性:(46.48±2.21)U/mg prot、(38.49±2.56)U/mg prot、(33.80±1.73)U/mg prot、(27.65±2.05)U/mg prot;GSH-Px活性:(21.41±1.95)U/mg prot、(17.32±1.28)U/mg prot、(15.09±0.94)U/mg prot、(14.08±1.23)U/mg prot;MDA水平:(1.41±0.09)nmol/mg prot、(1.59±0.09)nmol/mg prot、(1.81±0.09)nmol/mg prot、(2.16±0.14)nmol/mg prot;实验组SOD和GSH-Px酶活性均明显低于对照组(均P<0.05),而MDA水平则明显高于对照组(P<0.05)。对照组、低、中、高剂量组α-葡萄糖苷酶水平分别为(15.46±0.71)U/mL prot、(12.95±0.72)U/mL prot、(11.34±0.65)U/mL prot、(8.76±0.60)U/mL prot;LC分别为(6.21±0.31)μg/L、(5.89±0.13)μg/L、(5.02±0.12)μg/L、(4.38±0.07)μg/L,与对照组比较,实验组α-葡萄糖苷酶、LC浓度均明显下降(均P<0.01);对照组和低、中、高剂量组附睾Nrf2的表达水平分别为(1.34±0.05)ng/L、(1.25±0.04)ng/L、(1.08±0.06)ng/L、(0.92±0.04)ng/L,OCTN2的表达水平分别为(4.55±0.12)ng/L、(4.23±0.11)ng/L、(3.20±0.24)ng/L、(2.59±0.05)ng/L,与对照组比较,实验组Nrf2、OCTN2表达水平均明显下降(均P<0.01)。结论乙烯利暴露使大鼠生殖器官产生过量活性氧,发生氧化应激,可能通过激活Keap1-Nrf2/ARE信号通路,使其精子数量减少,活力降低,精子质量下降,导致生育能力受损。
Objective To investigate the effect of ethephon exposure on sperm quality of adolescent male SD rats and the influence mechanism.Methods A total of 4045-day-old male SD rats were divided into control group and low,middle and high experimental groups according to the random number table method,10 rats in each group.The said 4 groups were given 9 g/L normal saline,100 mg/kg,200 mg/kg,and 400 mg/kg ethephon aqueous solution for 28 days,respectively.One epididymal tail was taken to prepare sperm suspension,the sperm concentration and motility were detected.The testis and epididymis tissues were stained with HE,and their pathological changes were observed under light microscope.The activities of superoxide dismutase(SOD)and glutathione peroxidase(GSH-Px),and the malondialdehyde(MDA)content in the testis were detected.Enzyme linked immunosorbent assay kit was used to mea-sure the epididymalα-glucosidase activity,L-carnitine(LC)content,nuclear factor erythroid 2-related factor 2(Nrf2)and organic cation transporter 2(OCTN2)expression levels.Then the oxidative damage caused by ethephon to epididymis was evaluated.SPSS 26.0 software was used for data analysis.Data were compared by One-way ANOVA among groups and LSD method between 2 groups.Results The sperm concentration of the control group,low,medium and high dose groups were(40.21±1.94)×10^(9)/L,(35.23±2.53)×10^(9)/L,(23.61±2.62)×10^(9)/L,and(18.86±2.16)×10^(9)/L,respectively.The sperm activity rate were(70.98±3.01)%,(57.96±3.75)%,(45.71±2.41)%,and(31.23±2.26)%,respectively.The concentration and vitality of epididymal sperms in the experimental group were significantly lower than those in the control group(all P<0.01).In the control group,low,medium and high dose groups,the SOD activity were(46.48±2.21)U/mg prot,(38.49±2.56)U/mg prot,(33.80±1.73)U/mg prot,and(27.65±2.05)U/mg prot,respectively.The GSH-Px activity in said 4 groups were(21.41±1.95)U/mg prot,(17.32±1.28)U/mg prot,(15.09±0.94)U/mg prot,and(14.08±1.23)U/mg prot,respectively.The MDA content in said 4 groups were(1.41±0.09)nmol/mg prot,(1.59±0.09)nmol/mg prot,(1.81±0.09)nmol/mg prot,and(2.16±0.14)nmol/mg prot,respectively.Compared to the control group,the experimental groups had significantly lower SOD and GSH-Px activities and significantly higher MDA content(all P<0.05).α-glucosidase levels in the control group,low,middle and high experimental groups were(15.46±0.71)U/mL prot,(12.95±0.72)U/mL prot,(11.34±0.65)U/mL prot,and(8.76±0.60)U/mL prot,respectively.LC levels in the control group,low,middle and high dose groups were(6.21±0.31)μg/L,(5.89±0.13)μg/L,(5.02±0.12)μg/L,(4.38±0.07)μg/L,respectively,compared with those of the control group,the concentration ofα-glucosidase and LC in experimental groups decreased significantly(all P<0.01).The expression levels of Nrf2 in epididymis of the control group,low,middle and high dose groups were(1.34±0.05)ng/L,(1.25±0.04)ng/L,(1.08±0.06)ng/L,(0.92±0.04)ng/L,respectively;the expression levels of OCTN2 in epididymis of the control group,low,middle and high dose groups were(4.55±0.12)ng/L,(4.23±0.11)ng/L,(3.20±0.24)ng/L,(2.59±0.05)ng/L,respectively,compared with those of the control group,the expression levels of Nrf2 and OCTN2 in experimental groups decreased significantly(all P<0.01).Conclusions Ethephon exposure leads to excessive generation of reactive oxygen and oxidative stress in reproductive organs.Ethephon exposure may activate the Keap1-Nrf2/ARE signal pathway,resulting in a decrease in the number,vitality and quality of sperms,and impaired fertility.
作者
杨中华
宋翠萍
张海洋
饶旺
邵秋平
原志庆
Yang Zhonghua;Song Cuiping;Zhang Haiyang;Rao Wang;Shao Qiuping;Yuan Zhiqing(The First Clinical College of Xinxiang Medical University,Weihui 453100,China;Department of Pediatric Surgery,First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,China;Xinxiang Medical University,Xinxiang 453003,China)
出处
《中华实用儿科临床杂志》
CAS
CSCD
北大核心
2022年第23期1813-1817,共5页
Chinese Journal of Applied Clinical Pediatrics
基金
新乡市科技攻关项目(CXGG17006,GG2021031)。
关键词
乙烯利
氧化应激
精子质量
Ethephon
Oxidative stress
Sperm quality
