This study is aimed to analyse the cross-link between cyclin D1, cdk-4, p21, PCNA and DNA damage during different periods of reperfusion following experimental torsion in rats. Thirty mature male Wistar rats (N = 6) were used. Following 4 hr from torsion induction, the reperfusion was induced. Animals were subdivided into groups, including 4 hr torsion-induced (T1), 1 hr post-reperfusion (T2), 2 hr post-reperfusion (T3), 4 hr post-reperfusion (T4) and 8 hr post-reperfusion (T5) groups. The seminiferous tubules differentiation (TDI) and spermatogenesis indices were evaluated. The expressions of cyclin D1, cdk-4, p21 and PCNA were analysed using Reverse Transcriptase-PCR (RT-PCR). Moreover, the cyclin D1 + , cdk-4 + , p21 + and PCNA + cell numbers/mm 2 of tissue were assessed through immunohistochemical staining. The testicular DNA fragmentation was analysed using TUNEL assay and DNA ladder test. Observations demonstrated that reperfusion significantly increased (p < 0.05) up-regulated the expressions of cyclin D1, cdk-4 and PCNA. The animals in T5 group showed diminished expression of p21 and represented diminished DNA fragmentation versus T1 group.In conclusion, minimum 8 hr post-reperfusion is needed to re-initiate necessary expressions of cyclin D1, cdk-4 and PCNA to restore cell cycling machinery and ameliorate torsion-induced DNA damage. K E Y W O R D SDNA damage, germ cell, reperfusion, testis, torsion

Apoptosis and antioxidant mechanisms are pathways for the treatment of endometriosis (Endo). Rutin (Rtn) is an antioxidant flavonol that induces apoptosis. This study, for first time, was conducted to evaluate the effects of rutin on Endo through apoptosis and antioxidant mechanisms. The experimental Endo was induced in 24 rats and then the animals were subdivided into Endo-sole, 3000 and 6000 µg/kg rutin (Rtn-3000 and Rtn-6000) and vitamin C groups. After 4 weeks, the expression of Bcl2, Bax, anti Pro Caspase-9, cleaved Caspase-9, pro PARP, pro Cleaved PARP, Pro PARP, pro mTOR and mTOR were assessed by western blotting technique. The protein concentrations of malondialdehyde (MDA), total antioxidant capacity, and super oxide dismutase and gutathione peroxidase were also evaluated. TUNEL staining was also used for the detection of apoptosis. Caspase-9 and concentration of antioxidants were higher in the treated groups compared to Endo-sole group (P < 0.05). The results also showed that rutin decreased the expression of Bcl2 and MDA concentration (P < 0.05). The results for TUNEL staining showed that the animals treated with Rtn-6000 and vitamin C showed higher apoptosis. Rutin induces apoptosis by the expression of Bcl-2, Bax and caspase and also antioxidant activity by increasing antioxidants concentrations.

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