Cis-unsaturated fatty acids stimulate reactive oxygen species generation and lipid peroxidation in human spermatozoa.
BACKGROUND
Defective sperm function is the largest defined cause of human infertility; however, the etiology of this condition is poorly understood. Although oxidative stress is acknowledged as a key contributor to this pathology, there are also data indicating that defective human spermatozoa contain abnormally high amounts of cis-unsaturated fatty acids. This study investigated whether a causative relationship exists between these two attributes of impaired semen quality.


OBJECTIVE
The objective of this study was to determine whether polyunsaturated fatty acids can induce oxidative stress in human spermatozoa.


METHODS
Dihydroethidium and SYTOX Green were used in conjunction with flow cytometry and HPLC to investigate reactive oxygen species (ROS) generation by human spermatozoa after fatty acid exposure.


RESULTS
Arachidonic acid (AA) induced a time- and dose-dependent increase in ROS generation by human spermatozoa that led to the promotion of peroxidative damage and a loss of sperm motility. This effect could not be blocked with inhibitors of the cyclooxygenase or lipoxygenase pathways of AA metabolism, rotenone, protein kinase C antagonists, or known inhibitors of plasma membrane redox systems. However, ROS generation could be triggered with other cis-unsaturated fatty acids including linoleic and docosahexaenoic acids. Saturated fatty acids, methyl esters of unsaturated fatty acids, or other amphiphiles were all ineffective. However in a cell-free system, AA could trigger a redox signal via mechanisms that were profoundly disrupted by diphenylene iodonium, a flavoprotein inhibitor.


CONCLUSIONS
The presence of excess unsaturated fatty acids in defective human spermatozoa may precipitate the oxidative stress encountered in male infertility.
Aitken
R John
RJ
Discipline of Biological Sciences, School of Environmental and Life Sciences, University of Newcastle, Callaghan, New South Wales 2308, Australia. jaitken@mail.newcastle.edu.au
Wingate
Jordana K
JK
De Iuliis
Geoffry N
GN
Koppers
Adam J
AJ
McLaughlin
Eileen A
EA
D016718:Arachidonic Acid / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005231:Fatty Acids, Unsaturated / Q000494:pharmacology*; D006801:Humans; D015227:Lipid Peroxidation / Q000187:drug effects*; D008297:Male; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors; D011720:Pyrazoles / Q000494:pharmacology; D017382:Reactive Oxygen Species; D013094:Spermatozoa / Q000378:metabolism*
D005231:Fatty Acids, Unsaturated; D011720:Pyrazoles; D017382:Reactive Oxygen Species; C115461:SC 560; D016718:Arachidonic Acid; D019255:NADPH Oxidases