[Sperm mtDNA content and mtDNA4977bp deletion in normal and leukocytospermia men].
OBJECTIVE
To determine the sperm mtDNA content, mtDNA4977bp deletion and ROS in the seminal plasma of normal and leukocytospermia men, and to investigate the correlation of the changes of sperm mtDNA with the increase of leukocytes and reactive oxgygen species (ROS) in the seminal plasma.


METHODS
Seventy-eight semen samples from leukocytospermia patients and 31 from healthy donors were divided into 3 layers, supernatant fluid, 30% sperm and 80% sperm, by Percoll gradient centrifugation, their sperm mtDNA content and mtDNA4977bp deletion quantitatively analyzed by real-time PCR, and the level of ROS determined by flow cytometry.


RESULTS
The ROS in the seminal plasma and the sperm mtDNA contents of the three layers were all significantly higher in the leukocytospermia group than in the healthy control (P < 0.01). In the supernatant fluid and 80% layers, mtDNA4977bp deletion showed no obvious difference between the control and the leukocytospermia group, but was significantly higher in the 30% layer of the latter (P < 0.01). The ROS level was found positively correlated with the mtDNA content in the 30% (r = 0.347, P < 0.01) and the 80% layer (r = 0.456, P < 0.01), but not in the supernatant layer.


CONCLUSIONS
The increase of leukocytes and ROS may be one of the causes of the enhanced sperm mtDNA content, but has no significant impact on the mtDNA4977bp deletion.
Postgraduate Institute of Peking Union Medical College, Beijing 100730, China. zhouwanhao@163.com
Wan-Hao
WH
Zhou
Xu
X
Ma
Hui
H
Jiang
Ren-Pei
RP
Yuan
Qian
Q
Chen
Yu-Jie
YJ
Sui
Meng-Chun
MC
Jia
D000328:Adult; D004272:DNA, Mitochondrial / Q000235:genetics* / Q000378:metabolism*; D005434:Flow Cytometry; D006801:Humans; D007248:Infertility, Male / Q000235:genetics / Q000378:metabolism / Q000503:physiopathology*; D007962:Leukocytes / Q000737:chemistry / Q000378:metabolism; D007964:Leukocytosis / Q000235:genetics / Q000378:metabolism / Q000503:physiopathology; D008297:Male; D016133:Polymerase Chain Reaction; D017382:Reactive Oxygen Species / Q000378:metabolism; D017384:Sequence Deletion; D013076:Sperm Count; D013094:Spermatozoa / Q000166:cytology / Q000378:metabolism*
D004272:DNA, Mitochondrial; D017382:Reactive Oxygen Species