The Impact Of The Combination Of Low Dose Gossypol And Steroidal Hormones On Rat Testicular Cdc25a And Cyclin B1 And Ar Protein Expression

Adult male rats were randomly divided into four groups. Group GH:rats were fed orally with Gossypol acetic (GA,12.5mg/kg·day) and Desogestrel (DSG,125μg/ kg·day)/Ethinylestradiol (E,25μg/kg·day)/Testosterone undecanoate (TU, 100mg/ kg·day); Group G:a single dose of GA (12.5mg/kg·day) was given; Group H:DSG (125μg/kg·day)/E (25μg/kg·day)/TU (100mg/kg·day) were administered; Group C: rats only received vehicle (1% methyl cellulose). The testes were excised at the 6th,8th and 10th week, respectively. To investigate the influence of Cdc25A, cyclin B1 and androgen receptor (AR) by administering with combination regimen of low-dose gossypol acetic acid with steroid hormones (DSG/E/TU) in adult rat testis, the expression of Cdc25A, cyclin B1 and AR was detected by immunohistochemical staining and Western blotting.Immunohistochemistry results demonstrated that Cdc25A was expressed in the nuclei of spermatogonia and preleptotene spermatocytes, the cytoplasm of pachytene and diplotene spermatocytes as well as the acrosomes of round spermatids in the testis of control group. Compared with the control group, the expression of Cdc25A decreased in Leydig cells and acrosomes in GH group at the 6th,8th and 10th week (P<0.05). The expression of Cdc25A increased in spermatocytes in GH group at the 8th week (P<0.05). Immunohistochemistry results showed that cyclin B1 was expressed both in Leydig cells and round spermatids in the testis of control group. However, the expression of cyclin B1 decreased in Leydig cells in GH group (P<0.05). Immunohistochemistry results demonstrated that AR was expressed in the nuclei of leptotene and zygotene spermatocytes, the head of step 11 spermatids and the cytoplasm of Leydig cells. However, the AR positive staining appeared in the nuclei of Leydig cells in GH group.Western blotting showed that the expression of Cdc25A in rat testis increased in GH group with treatment time increasing, opposite to the control group; the expression of cyclin B1 in rat testis in GH group is similar with the control group.The different expression of Cdc25A in spermatocytes and spermatids in normal rat testis suggests that Cdc25A may play important roles at meiotic phase in spermatogenesis. High-level expression of Cdc25A in acrosomes may be related to the formation of acrosomes. The effects of low-dose gossypol acetic acid with steroid hormones on the expression of Cdc25A were different, which indicate that there is a potential influence on Cdc25A's function at meiotic phase in spermatogenesis. The changes of Cyclin B1 in Leydig cells in GH group suggest that combination regimen of low-dose gossypol acetic acid with steroid hormones inhibits the function of Leydig cells, which do not make direct impact on spermatogenesis. By detection of the expression of AR, we found that combination regimen of low-dose gossypol acetic acid with steroid hormones did not exert an immediate impact on the function of AR in leptotene and zygotene spermatocytes, instead of the functional status of AR in Leydig cell.