生殖医学杂志2018，Vol.27Issue(5)：451-459,9.DOI:10.3969/j.issn.1004-3845.2018.05.011
LonP1表达与线粒体稳态以及精子活力的关联性研究
Relationship between LonP1 expression and sperm mitochondrial homeostasis & sperm motility
摘要
Abstract
Objective:To investigate the role of mitochondrial lon protein hydrolase(LonP1) in the maintenance of sperm mitochondrial homeostasis and its possible regulatory mechanisms.Methods:The semen specimens of 120 male donors from Reproductive Center of First Affiliated Hospital of Wenzhou Medical University during Jun.2016 to Jun.2017 were collected,and then the semen specimens were divided into two groups:normal viable spermatozoa(NS) and asthenospermia(AS).The semen parameters of each group were detected by computer assisted semen analysis system(CASA).The expression of LonP1 was determined by Western blotting and immunofluorescence.The mitochondrial membrane potential(MMP) (In terms of the ratio of green/red fluorescence intensity,the smaller the ratio,the higher the membrane potential) and mitochondrial reactive oxygen species (mROS) of sperm were determined by flow cytometry.Then the correlation between LonP1 expression,sperm motility and sperm mitochondrial function-related indicators(MMP,mROS) were analyzed.In addition,semen in NS group was treated by density gradient centrifugation method,and the precipitated sperms were re-suspended in culture medium.Sperms were in vitro culture treated with the final concentrations (2.5 μmol/L and 5 μmol/L) of LonP1 inhibitor CDDO-ME.Meanwhile,the blank group(0 μmol/L) was set.The sperm motility parameters,LonP1 expression level and sperm mitochondrial function-related indexes were compared among the groups.Results:LonP1 was stably expressed in the mitochondrial sheath in the middle part of sperm in human mature sperm.The expression level of LonP1 in NS group and AS group(0.52 vs.0.22) was positively correlated with MMP(4.72％ vs.12.73％) and sperm motility(69.89％ vs.26.10％) (P＜0.05),and negatively correlated with the mROS levels(418.85 vs.460.52)(P＜0.05).The expression of LonP1 protein was significantly decreased after the treatment of CDDO-ME(P＜0.05).The expression of LonP1 protein in CDDO-ME treatment group was significantly decreased with the increase of CDDO-ME concentration(P＜0.05).Meanwhile,MMP was significantly decreased (78.81％ vs.7.34％),and the mROS level was significantly increased(640.68 vs.462.26) in 5 μmol/L group compared with the blank group(P＜0.05).After CDDO-ME treatment,the semen parameters VAP and VSL were significantly decreased with the concentrations of CDDO-ME(P＜0.05).The PR(52.08％ vs.65.15％),VCL(48.28μm/s vs.78.78 μm/s),ALH(1.28 μm vs.1.93 μm),LIN(38.83％ vs.56.05％) and STR(45.53％ vs.81.25％) were significantly decreased in 5 μmol/L group(P＜0.05).Conclusions:LonP1 was stably expressed in the mitochondrial sheath in the middle part of human mature sperm.LonP1 can improve sperm motility by maintaining sperm mitoehondrial homeostasis.关键词
精子/LonP1蛋白/线粒体/CDDO-MEKey words
Sperm/LonP1 Protein/Mitochondria/CDDO-ME引用本文复制引用
张家燕,王晓娜,李金晶,黄小圆,朱春芳,费前进,葛红山..LonP1表达与线粒体稳态以及精子活力的关联性研究[J].生殖医学杂志,2018,27(5):451-459,9.基金项目
国家自然科学基金(2016-81501312) （2016-81501312）
