Study On Semen Quality And Sperm Genetic Damage Of Adult Male In The Area Of Three Gorges Reservoir

A lot of data and research show that human reproductive function is degraded gradually. These studies also indicated semen quality may change along with the geographical differences, which may be caused by exposure to chemicals in the environment, living habits and some other unknown factors. The world-famous Three Gorges Project is the largest project to bring benefit to the people. At the same time, it inevitably leads to changes in the ecological environment. Thus, it is necessary to comprehensively understand the status of male reproductive health and explore the risk factors of semen quality.The objectives of our research include three aspects ,①to understand the semen quality and potential factors of healthy adult men in the Three Gorges Reservoir Region,②to observe the DNA damage in sperm,③to determin the metabolites of polycyclic aromatic hydrocarbons (PAHs) in human urine and to explore the relationshiop between PAHs metabolites and genetic damage of sperm.We Selected 3 districts and 3 counties which were representatives of the Three Gorges Reservoir Region and collected semen specimens. We analysed semen quality by computer-assisted semen analysis system (CASA), and detected sperm DNA damage by single cell gel electrophoresis (SCGE). Using high-performance liquid chromatography (HPLC) with fluorescence detection (FLD) , we detected metabolites of PAHs in urinary, including 2-hydroxynaphthalene(2-OHNa), 2-hydroxyfluorene(2-OHFlu), 9-hydroxyphenanthrene (9-OHPh) and 1-hydroxypyrene (1-OHP), and tried to find the correlations between sperm DNA damage and levels of metabolites of PAHs. The results of our research are as follows:①Of the 1346 semen samples evaluated, only 524(38.9%) had all normal semen parameters according to WHO criteria, and 61.1% had at least one of the semen parameters below normal threshold values. Season and abstinence duration were found to be significantly associated with semen quality.②The mean values of semen volume and total motility in our study were in agreement with other studies done in Chinese men, but the mean sperm concentration and total sperm count in our study (84.8×10~6/ml and 203.2×10~6/ml, respectively) were markedly higher than those regions, ranging from 55.9 to 64.5×10~6/ml, and from 133.6 to 164.2×10~6/ml, respectively. The semen quality of adult male in the Three Gorges Reservoir Region is better than other regions. ③Experiment platform of sperm SCGE was established by our laboratory. The best conditions of our laboratory for this experiment are as follows: pyrolysis time 60 min, proteinase K digestion time of 15 ~ 18h, the time for helicase 20 min, voltage 22V, current 260 mA, electrophoresis time 5 min.④The minimum concentrations of 2-OHNa, 2-OHFlu, 9-OHPh and 1-OHP in urine of 219 adult male in Chongqing area are 0.13,0.44,0.82,0.44ug/L; the highest concentrations are 65.6, 222, 412, 218 ug/L. By corrected by creatinine (CR) concentration, the median of concentration (ug/g.CR) are 8.45, 3.05, 2.01 and 0.70, respectively. The concentration of 2-OHNa, 2-OHFlu, 9-OHPh and 1-OHP in adult male urine of Chongqing have similar results of studies in China, but higher than other countries.⑤2-OHNa was positively correlated with tail DNA%, tail length and tail moment respectively; 2-OHFlu was positively correlated with tail DNA%; 1-OHP was positively correlated with the tail DNA% and tail moment (P <0.01). 9-OHPh has no correlation with the sperm DNA damage. The metabolites of polycyclic aromatic hydrocarbons in urine have correlation with sperm DNA damage, suggesting that polycyclic aromatic substances might cause male reproductive damage.