[Experimental study of the effect of rhTNF-alpha on human sperm mitochondrial function and motility in vitro].
OBJECTIVE
To study the effect of rhTNF-alpha on human sperm mitochondrial function and motility in vitro.


METHODS
Fifty-six semen samples collected by masturbation were analyzed according to WHO protocols. Semen samples from 40 healthy men were prepared using Percoll centrifugation. Sperm suspension was diluted to a concentration of 10 x 10(6)/ml in Ham's F10 medium. Sperm samples were incubated with rhTNF-alpha solution (final concentration 0.03 microg/L, 0.06 microg/L, 0.09 microg/L and 0.27 microg/L, respectively) for 0.5 h, 1 h, 2 h, 3 h and 4 h at 37 degrees C in 5% CO2, and comparative studies were made with a control group. Ten microl sperm samples were examined with CASA technique, 250 microl stained in the presence of 10 microg/ml Rh123 and PI, and mitochondrial function analyzed by flow cytometry.


RESULTS
Significant differences were found between the experimental groups (final concentration 0.06 microg/L, 0.09 microg/L and 0.27 microg/L) and the control group in viability, straight line velocity, curvilinear velocity, average path velocity, progressive motility of human sperm and the number of spermatozoa with normal mitochondrial function (P < 0.01) except the final concentration 0.03 microg/L group (P > 0.05). Motility of human sperm lowered with the increase of rhTNF-alpha concentration and incubation time, and r values were 0.675, 0.691, 0.762, 0.693, 0.724 and 0.571, 0.594, 0.752, 0.791, 0.816, respectively (P < 0.01). The number of spermatozoa with normal mitochondrial function decreased with the increased rhTNF-alpha concentration and incubation time, and r values were 0.615, 0.643, 0.752, 0.691, 0.754 and 0.532, 0.567, 0.782, 0.692, 0.854, respectively (P < 0.01).


CONCLUSIONS
rhTNF-alpha can reduce human sperm motility function in vitro, possibly by interfering with human sperm mitochondrial function.
Bian
Jiang
J
Research Institute of Andrology, Medical College, Three Gorges University, Yichang, Hubei 443003, China. bianjiang2002@yahoo.com.cn
Guo
Xiankun
X
Xiong
Chengliang
C
Li
Jiahua
J
Tian
Yonghong
Y
Ma
Huagang
H
Zhang
Yan
Y
Nie
Yong
Y
Yu
Lingli
L
Xiao
Li
L
D000328:Adult; D004305:Dose-Response Relationship, Drug; D006801:Humans; D008297:Male; D008928:Mitochondria / Q000187:drug effects* / Q000502:physiology; D011994:Recombinant Proteins / Q000494:pharmacology; D013081:Sperm Motility / Q000187:drug effects*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha