The effect of alpha lipoic acid on testical damage caused by busulfan on prepubertal rats

Objective: In this study we aimed to investigate protective effects of Alpha Lipoic Acid, a strong antioxidant, against possible negative effects of busulfan, on testes of male rats histolochemically, imunohistochemically and biochemically. Method: 21 Wistar albino rats were divided into 4 groups (n=7). I.control goup: rates were givin normal saline orally for 35 days. II.Busulfan group : rates were given busulfan twice daily for 21 days in a dose of 10mg/kg , intraperitoneally. III.alpha lipoic acid group: rates in this group were given alpha lipoic acid orally in a dose of 35mg/kg for 5 weeks. IV.alpha lipoic acid + busulfan group: in this group rates were given the fisrt dose of busulfan in a dose of 10mg/kg on the first day . on the seconde day alpha lipoic acid started to be given in adaily dose of 35mg/kg for 5 weeks. the seconde dose of busulfan was given in a dose of 10mg/kg on 21.day of the experiment. At the end of the experiment, all subjects were sacrificed. the left testests of the rates were prepared for hematoxylene & eosine (H&E), periodic acid schiff (PAS), TUNEL and active caspase-­3 painting. The right testeses of the rates were evaluated biochemically by Glutathion (GSH), glutathion peroxidase (GPx) and malondialdehyde (MDA). Result: According to histologic and morphometric evaluation on testicles, it is observed that Busulfan group is damaged when compared to control and Alpha Lipoic Acid group, however in ALA+BUS group Alpha Lipoic Acid is not able to prevent the damage. The positive cells are counted immunohistochemically with Tunel and Active Caspas-­3 coloring. It is observed that the apoptotic cell amount has risen significantly in Busulfan and ALA+BUS groups when compared to the control group. It is also observed that there is no significant difference between Busulfan and ALA+BUS groups in terms of apoptotic cell amount. The measurements have been conducted biochemically by MDA GPx and GSH kits. MDA values are found significantly higher in busulfan group than in control group and Alpha Lipoic Acid group. No significant difference is observed between control and Alpha Lipoic Acid groups. Compared to Busulfan group, the MDA value is found significantly high in ALA+BUS group. GSH values are found significantly low in Busulfan group when compared to control group. And when compared to control group, GSH values are found significantly high in Alpha Lipoic Acid. GSH values are found significantly high in alabus group when compared to Busulfan group. GPx values are found significantly low in Busulfan group when compared to control group. It has been also observed that there is no significant difference between control group alpha lipoic acid group and ALA+BUS group. When compared to Busulfan group, GPx value is found significantly high in ALA+BUS group. Conclusion: According to our results, it has been been observed that Busulfan rises apoptose and causes testicle damage while Alpha Lipoic Acid has no protective/curing effect on testicle damage when used with Busulfan. Key Word: Busulfan, Alpha Lipoic Acid, Testis