Determination of testicular injury in mouse with azospermia via serum miRNA level

CAim: Cyclophosphamide (CP) is an alkylating, chemotherapeutic agent commonly used in cancer treatments and has a detrimental effect on many organs, especially gonads. In this study, we aimed to determine the relationship between GDNF, Okludin and TGF-β3 primary antibodies determined by immunohistochemical techniques and mir-34b and mir-34c in blood and tissue samples in testicular injury after CP administration in mice. Materials and Methods: Eight weeks old male rats (n: 18) from Manisa Celal Bayar University Laboratory of Experimental Animals Research and Application Center were divided into three groups. No treatment was applied to the mice in the control group. Mice in the experimental group were injected 100 mg/kg CP intraperitoneally once daily for 5 days, and in the sham group the same amount of saline was injected. 24 hours after the last injection, the animals were sacrificed, and testicular tissues and blood samples were taken. Tissues and blood samples were analyzed by immunohistochemical and RT-PCR methods. Results: Immunohistochemical analysis of testicular tissues showed a decrease in both spermatogenic germ cell count and GDNF and Okludin expressions, and an increase in TGF-β3' expression in the CP group compared to the control and sham group. When the expression of mir-34b and mir-34c were examined by RT-PCR technique, significant decrease was observed in tissue samples in the CP-treated group, but sufficient expression was not observed in plasma samples of the all groups. Conclusion: In this study, it was concluded that the expression of GDNF, Okludin and TGF-β3 plays an important role in testicular injury caused by CP, and the decrease in the expression of mir-34b and mir-34c in tissue samples may be an important marker for the detection of testicular damage, however, further investigations are needed to detect them in plasma. Keywords: Cyclophosphamide, testes, GDNF, Okludin, TGF-β3, miR34b/c