Increase of oxidative stress in human sperm with lower motility.
OBJECTIVE
To investigate the causal role of oxidative-stress status on human sperm motility.


METHODS
To demonstrate that sperm with higher oxidative damage have a lower antioxidant capacity.


METHODS
University hospital infertility center.


METHODS
Seventy-eight semen samples were obtained from 35 healthy donors who had normal semen characteristics and from 43 infertile or subfertile males.


METHODS
The levels of oxidative damage (8-hydroxy-2'-deoxyguanosine [8-OHdG] and lipid peroxides) and antioxidants (retinol, alpha-tocopherol, ascorbate, and protein thiols) in the spermatozoa and/or seminal plasma were measured.


METHODS
We analyzed the specific content of 8-OHdG and lipid peroxides by using high-performance liquid chromatography (HPLC)-electrochemical detection and HPLC-fluorescence analysis, respectively. Retinol and alpha-tocopherol were analyzed by using an HPLC system, whereas ascorbate and protein thiols were determined by using spectrophotometry. 8-Hydroxy-2'-deoxyguanosine was visualized by immunofluorescent staining with an anti-8-OHdG antibody that was conjugated with fluorescein isothiocyanate conjugate. Lipid peroxides in spermatozoa were stained with a fluorescent dye, C11-BODIPY(581/591).


RESULTS
Statistically significant negative correlations were revealed between sperm motility and 8-OHdG and between motility and lipid peroxides. Statistically significant positive correlations were found between sperm motility and the levels of retinol, alpha-tocopherol, ascorbate, and protein thiols of seminal plasma. 8-Hydroxy-2'-deoxyguanosine and lipid peroxides in spermatozoa were found to be present mostly in mitochondria.


CONCLUSIONS
Oxidative stress and oxidative damage were increased significantly in spermatozoa with declined motility, and the antioxidant capacities in the spermatozoa and seminal plasma were lower in males who had infertility or subfertility.
School of Medical Laboratory Science and Biotechnology, Taipei Medical University, Taipei, Taiwan; Department of Biochemistry and Molecular Biology, National Yang-Ming University, Taipei, Taiwan.
Shu-Huei
SH
Kao
Department of Obstetrics and Gynecology, Taipei Veterans General Hospital, Taipei, Taiwan.
Hsiang-Tai
HT
Chao
School of Nutrition, Chung-Shan Medical University, Taipei, Taiwan.
Haw-Wen
HW
Chen
Department of Urology, Hsin Kong Wu Ho-Su Memorial Hospital, Taipei, Taiwan.
Thomas I S
TIS
Hwang
School of Medical Laboratory Science and Biotechnology, Taipei Medical University, Taipei, Taiwan.
Tien-Ling
TL
Liao
Department of Biochemistry and Molecular Biology, National Yang-Ming University, Taipei, Taiwan. Electronic address: joeman@ym.edu.tw.
Yau-Huei
YH
Wei
D000080242:8-Hydroxy-2'-Deoxyguanosine; D000975:Antioxidants / Q000378:metabolism*; D001205:Ascorbic Acid / Q000378:metabolism; D004249:DNA Damage / Q000502:physiology; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000378:metabolism; D006801:Humans; D007248:Infertility, Male / Q000378:metabolism / Q000503:physiopathology*; D008054:Lipid Peroxides / Q000378:metabolism; D008297:Male; D008928:Mitochondria / Q000378:metabolism; D018384:Oxidative Stress / Q000502:physiology*; D017382:Reactive Oxygen Species / Q000378:metabolism; D012661:Semen / Q000378:metabolism; D013081:Sperm Motility / Q000502:physiology*; D013094:Spermatozoa / Q000378:metabolism*; D013438:Sulfhydryl Compounds / Q000378:metabolism; D014801:Vitamin A / Q000378:metabolism; D024502:alpha-Tocopherol / Q000378:metabolism
D000975:Antioxidants; D008054:Lipid Peroxides; D017382:Reactive Oxygen Species; D013438:Sulfhydryl Compounds; D014801:Vitamin A; D000080242:8-Hydroxy-2'-Deoxyguanosine; D003849:Deoxyguanosine; D024502:alpha-Tocopherol; D001205:Ascorbic Acid