[Effects of p,p'-DDE and beta-BHC on the apoptosis of Sertoli cells in vitro].
OBJECTIVE
To explore the effects of p,p'-DDE and beta-BHC on the apoptosis of Sertoli cells in vitro via activation of Caspase.


METHODS
Sertoli cells were treated in vitro for 24 hours with a serial concentrations of p,p'-DDE (10, 30 and 50 micromol/L), beta-BHC (10, 30 and 50 micromol/L) and p,p'-DDE + beta-BHC (10, 30 and 50 micromol/L). The inhibitory group was first treated with 100 micromol/L Caspase-3 inhibitor Ac-DEVD-CHO treating for 2 hours before 50 micromol/L p, p'-DDE + 50 micromol/L beta-BHC 24 hours-treatment. The vitality of Sertoli cells was determined by MTT and the apoptosis rate was measured by AO/EB double fluorescence staining. The expressions of Caspase-3, Caspase-8 and Caspase-9 were determined by RT-PCR.


RESULTS
Average optical density (A) values were 0.498 +/- 0.039, 0.481 +/- 0.065, 0.397 +/- 0.032 and 0.286 +/- 0.049 in p,p'-DDE groups (10, 30, 50 and 70 micromol/L), and 0.518 +/- 0.103, 0.490 +/- 0.060, 0.454 +/- 0.054 and 0.302 +/- 0.030 in beta-BHC groups (10, 30, 50 and 70 micromol/L). In the mixture-treated groups (10, 30 and 50 micromol/L), the average A values were 0.483 +/- 0.048, 0.473 +/- 0.058 and 0.337 +/- 0.052. Compared with the solvent control group (0.527 +/- 0.022) , 50 micromol/L group of p, p'-DDE, beta-BHC or their mixture caused a significant decrease of Sertoli cell viability (t values were 4.599, 2.716, 6.537 respectively, P < 0.05). AO/EB double fluorescence staining analysis showed that apoptosis rates of Sertoli cells were significantly increased with all treated groups. The expressions of Caspase-3, Caspase-8 and Caspase-9 were upregulated as the concentrations of p,p'-DDE, beta-BHC and their mixture were increased.


CONCLUSIONS
p,p'-DDE, beta-BHC and their mixture could induce the apoptosis of Sertoli cells in vitro which was associated with activation of Caspase-3 mediated by cleavage of Caspase-8 and Caspase-9.
Liang
Xian-Min
XM
Department of Occupational and Environmental Health and MOE Key Laboratory of Environment and Health, School of Public Health,Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, China.
Hu
Ya-Fei
YF
Yu
Hai-Ge
HG
Yang
Ke-Di
KD
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D053181:Caspase 8 / Q000378:metabolism; D053453:Caspase 9 / Q000378:metabolism; D002478:Cells, Cultured; D004000:Dichloroethylenes / Q000633:toxicity*; D001556:Hexachlorocyclohexane / Q000633:toxicity*; D008297:Male; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D012708:Sertoli Cells / Q000187:drug effects* / Q000378:metabolism*
D004000:Dichloroethylenes; D012333:RNA, Messenger; D001556:Hexachlorocyclohexane; D053148:Caspase 3; D053181:Caspase 8; D053453:Caspase 9