Effects of p,p'-DDE and beta-BHC on the apoptosis of Sertoli cells in vitro / 中华预防医学杂志

<p><b>OBJECTIVE</b>To explore the effects of p,p'-DDE and beta-BHC on the apoptosis of Sertoli cells in vitro via activation of Caspase.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>

Action mechanism of p,p'-DDE and/or beta-BHC on JNK and MAPK signal transduction pathways in rat Sertoli cells / 中华男科学杂志

<p><b>OBJECTIVE</b>To study the action mechanism of p,p'-DDE and/or beta-BHC on JNK and MAPK signal transduction pathways in rat Sertoli cells in vitro.</p><p><b>METHODS</b>We cultured the Sertoli cells isolated from rat testicular tissues for 2 days in vitro, divided them into a control group incubated with DMSO and 3 case groups exposed to p,p'-DDE and / or beta-BHC at the final concentration of 10, 30, 50 micromol/L for 24 hours, and then detected the expression levels of JNK and c-jun mRNA by two-step RT-PCR.</p><p><b>RESULTS</b>Twenty-four hours after p,p'-DDE treatment, the grayscale values of JNK mRNA were 0.068 +/- 0.001, 0.164 +/- 0.002, 0.207 +/- 0.006 and 0.499 +/- 0.017, and those of c-jun mRNA were 0.122 +/- 0.002, 0.157 +/- 0.006, 0.218 +/- 0.007 and 0.289 +/- 0.004 respectively in the DMSO control and the 10, 30 and 50 micromol/L groups. The expressions of JNK and c-jun mRNA were elevated with increased concentration of p,p'-DDE, with significant differences between the control and the case groups (P < 0.05), and they were also significantly upregulated in the beta-BHC and p,p'-DDE + beta-BHC groups in a dose-dependent manner. The grayscale values of JNK mRNA in the p,p'-DDE, beta-BHC and p,p'-DDE + beta-BHC groups at the concentration of 10 micromol/L were 0.164 +/- 0.002, 0.149 +/- 0.003 and 0.178 +/- 0.004, and those of c-jun mRNA were 0.157 +/- 0.006, 0.131 +/- 0.004 and 0.172 +/- 0.002, respectively, both significantly higher in the combination group than in the former two (P < 0.05). And the same was the case with the 30 and 50 micromol/L concentrations.</p><p><b>CONCLUSION</b>Both p,p'-DDE and beta-BHC can enhance the expressions of JNK and c-jun in Sertoli cells, and their combination can produce even more obvious effect.</p>