白藜芦醇通过降低氧化应激改善过氧化氢诱导精子活力和DNA损伤  

Resveratrol improves hydrogen peroxide-induced sperm activity and DNA damage by reducing oxidative stress

作　　者：吴海峰[1] 李曼[1] 李玉静[1] 江乾[2] 李亮 赵世彬[2] 乜照燕[2] Wu Haifeng;Li man;Li yujing;Jiang Qian;Li Liang;Zhao Shibin;Nie Zhaoyan(Department of Laboratory Medicine,Hebei Chest Hospital,Shijiazhuang 050021 China;Department of Reproductive Medicine,The Fourth Hospital of Hebei Medical University,Shijiazhuang 050011,China)

机构地区：[1]河北省胸科医院检验科,河北石家庄050021 [2]河北医科大学第四医院生殖医学科,河北石家庄050011

出　　处：《中国男科学杂志》2021年第6期24-28,共5页Chinese Journal of Andrology

基　　金：河北省民生科技专项,192777106D;河北省重点科技研究计划(No.ZD20140140)。

摘　　要：目的探讨白藜芦醇(Resveratrol,RES)通过降低氧化应激改善过氧化氢(Hydrogen peroxide,H_(2)O_(2))诱导的精子活力和DNA损伤。方法收集我院生殖医学科精液检查正常标本,采用Percoll密度梯度离心法进行处理,处理后调整精子浓度为20×10^(6)/ml放置在5%CO_(2),37℃培养箱。不同浓度(0、25、50、75、100μmol/L)H_(2)O_(2)处理24 h,通过前向运动精子和总活力精子筛选H_(2)O_(2)浓度,建立H_(2)O_(2)精子损伤模型。进一步用不同浓度(0、10、25、50、75、100μmol/L)RES和H_(2)O_(2)共同作用24 h,通过前向运动精子和总活力精子选择RES浓度。根据上述确定RES和H_(2)O_(2)浓度,将实验分为三组:对照组(CON)、H_(2)O_(2)组(H_(2)O_(2))、H_(2)O_(2)+RES组(RES+H_(2)O_(2)),对三组精子处理后用伊红-苯胺黑染色检测精子膜完整性,硫代巴比妥酸法(TBA)测定MDA水平,精子吖啶橙染色检测精子DNA损伤。结果随H_(2)O_(2)浓度增加,前向运动精子和总活力精子降低,呈剂量依赖关系,选定100μmol/L H_(2)O_(2)为后续实验药物浓度。在H_(2)O_(2)精子损伤模型中随RES浓度增加,前向运动精子和总活力精子增加,50μmol/L RES对精子损伤改善效果最佳,确定为后续实验药物浓度。三组结果显示:H_(2)O_(2)组与CON组相比精子膜完整性降低,MDA水平上升,精子DNA损伤增加,而H_(2)O_(2)+RES组与H_(2)O_(2)组相比精子膜完整性提高,MDA水平下降,精子DNA损伤降低。结论RES可降低H_(2)O_(2)导致精子氧化损伤。Objective To investigate the effects of resveratrol on hydrogen peroxide-induced sperm activity and DNA damage.Methods The normal semen specimens were collected and processed by percoll density gradient centrifugation.The sperm concentration was adjusted to reach 20×10^(6)/ml,and treated with H_(2)O_(2)(0,25,50,75,100μmol/L)for 24 h in 5%CO_(2)37℃incubator,and the optimized concentration of H_(2)O_(2)was determined by progressive motility and total motility sperm to establish a model of H_(2)O_(2)injury.Subsequently,RES(0,10,25,50,75,100μmol/L)and H_(2)O_(2)were used together to treat sperm for 24 h,and the optimal concentration of RES was defined.The experiment was divided into three groups including the control group(CON),H_(2)O_(2)group(H_(2)O_(2))and RES+H_(2)O_(2)group(RES+H_(2)O_(2)).Eosin-nigrosine staining was used to detect sperm membrane integrity,thiobarbituric acid method(TBA)was used to detect MDA,acridine orange fluorescent staining was used to detect sperm DNA damage.Results With the increase of H_(2)O_(2),progressive motility and total motility sperm were decreased in a dose-dependent manner.100μmol/L H_(2)O_(2)was selected as the subsequent experimental drug concentration.Sperm treated with RES(0,5,10,25,50,and 75μmol/L),the progressive motility and total motility sperm were all increased.50μmol/L RES is determined as experimental drug concentration.Compared with that of the CON group,the sperm membrane integrity in H_(2)O_(2)group was decreased,the MDA and the sperm DNA damage were all increased.Compared with that in the H_(2)O_(2)group,the sperm membrane integrity in H_(2)O_(2)+RES group was increased,the MDA and the sperm DNA damage were decreased.Conclusion Resveratrol can reduce the oxidative stress damage of sperm caused by H_(2)O_(2).

关 键 词：白藜芦醇 过氧化氢 精子 氧化应激 精子DNA损伤 

分 类 号：R715.9[医药卫生—妇产科学] R698.2[医药卫生—临床医学]