摘要
目的探讨白藜芦醇(RES)对环磷酰胺诱导的人精子氧化损伤的保护作用。方法收集2018年1月至4月在河北医科大学第四医院生殖医学中心就诊男性患者的正常精液标本。将用密度梯度离心法处理后的正常精液标本用不同浓度(0、1.0、2.5、5.0、7.5μmol/L)环磷酰胺活性代谢产物4-过氧化氢环磷酰胺(4-HC)对精子进行体外毒性试验,继续培养24 h,通过精子活力和精子存活率筛选4-HC作用半抑制浓度(IC50)。将处理后的正常精液用不同浓度RES(5、10、25、50、75μmol/L)预处理2 h,再加入IC50浓度4-HC,测定精子活力和精子存活率选择RES最佳保护浓度。选择4-HC和RES浓度后,该实验分为四组:对照组(CON)、环磷酰胺组(4-HC)、环磷酰胺+白藜芦醇组(4-HC+RES)、白藜芦醇组(RES),检测各组精子活力、活率、氧化应激指标以及精子凋亡指标。结果随4-HC浓度增加,精子活力和存活率降低,呈剂量依赖关系,依据IC50,2.5μmol/L 4-HC为后续实验药物浓度。RES预处理后,精子活力和精子活率均得到改善,50μmol/L RES效果最佳,确定为后续实验药物浓度。结果显示:4-HC组较CON组,精子活力和存活率下降,氧化应激水平上升,精子凋亡增加,4-HC+RES组与4-HC组相比精子活力和存活率升高,氧化应激水平降低,精子凋亡降低。结论RES可以减少化疗药物导致的精子氧化应激水平增加,能预防和缓解化疗药物对人精子造成的损伤。
Objective To investigate the protective effect of resveratrol(RES)on the oxidative damage on human sperm induced by cyclophosptamide(Cy).Methods Normal semen samples were collected from the Department of Reproductive Medicine of the Fourth Hospital of Hebei Medical University during Jan.and Apr.2018.The samples were treated with density gradient centrifugation and cultured for 24 hours with different concentrations of 4-cyclophosphamide(4-HC)(0,1.0,2.5,5.0,7.5μmol/L).The sperm motility and survival rate were analyzed to confirm the IC50 of 4-HC.The samples were then pretreated for 2 h with different concentrations of RES(5,10,25,50,75μmol/L),and 4-HC with IC50 was added,the optimal pretreatment concentration of RES was selected.After that,the samples were divided into 4 groups:control group,4-HC group,4-HC+RES group,and RES group.The sperm motility,survival rate,oxidative stress indicators and apoptosis were detected.Results With the increase of 4-HC concentration,sperm motility and survival rate decreased in a dose-dependent manner.In line with IC50,2.5μmol/L was the concentration of 4-HC in subsequent experiments.After RES pretreatment,sperm motility and survival rate increased,and 50μmol/L RES was the optimal concentration.Compared with the control group,the 4-HC group had decreased sperm survival rate,but increased oxidative stress and apoptosis.Compared with the 4-HC group,4-HC+RES group had increased sperm survival rate but decreased oxidative stress and apoptosis.Conclusion RES can alleviate the damage on sperm by reducing the increased oxidative stress caused by chemotherapy drugs.
作者
乜照燕
张亚楠
花蕊
陈丽君
李亮
吴海峰
NIE Zhaoyan;ZHANG Ya nan;HUA Rui;CHEN Lijun;LI Liang;WU Haifeng(Department of Reproductive Medicine,The Fourth Hospital of Hebei Medical University,Shijiazhuang 050011;Department of Histology and Embryology,Hebei Medical University,Shijiazhuang 050010;Department of Laboratory,Hebei Chest Hospital,Shijiazhuang 050061,China)
出处
《现代泌尿外科杂志》
CAS
2021年第4期340-344,共5页
Journal of Modern Urology
基金
河北省民生科技专项(No.192777106D)
河北省重点科技研究计划(No.ZD20140140)。
关键词
白藜芦醇
环磷酰胺
精子
生殖毒性
氧化应激
凋亡
resveratrol
cyclophosptamide
sperm
reproductive toxicity
oxidative stress
apoptosis
