Ultrastructural and immunohistochemical examination of the protective effects of various antioxidants on chemotherapy applied rat ovarium follicules

One of the most common chemotherapeutic agents that is used in cancer treatment is cyclophosphamide; that toxic effects on female reproductive organs and causes fertilization failure. In our study, we aimed to immunohistochemically and electron microscobically evaluate the possible antioxidant effects of ascorbic acid, ?-tocopherol and selenium, on the degenerative effects of cyclophosphamide on normal ovarian tissue. To this end we used TGF-ß1, GDF-9, PCNA as developmental signal molecules; Caspase-3, APAF-1 as apoptotic signal molecules. TUNEL method is used in order to evaluate the DNA damage caused at the cell level.Following cyclosphamide application, we observed a decrease in TGF-ß1, PCNA immunoreaction in the granulosa cells of developing follicules and a decrease in GDF-9 immunoreaction in oocytes and a distinctive increase in caspase-3 and APAF-1 immunoreaction when compared to control group. Besides an increase in athretic follicules and in some Graaf follicules?s cells with DNA damage had showed a strong immunopositive reaction when compared to control group in TUNEL analysis. TGF-ß1 and PCNA immunoreactivity of developing follicules and GDF-9 immunoreaction of oocytes was similiar to the control group whereas Caspase-3 and APAF-1 immunoreaction in TUNEL analysis showed a distinctive decrease in both cyclosphamide and antioxidant treated group. Again in the same group the number of athretic folicules have found out to decrease significantly.Following electron microscobic examinations, we have observed intense degenerative changes in the multi layer primary follicles?s granulosa cells in cyclophosphamide treated group, which is an alike finding with immunohistochemical part of the study. In all of the three antioxidant treated groups, both ovaries and zona pellucida of the oocytes remained the same. Degeneration at the granulosa cells was moderately inhibited and there was quite a few decrease in the number of cells with dark cytoplasm.According to our observations, we have concluded that, degenerative changes at the ovaries caused by the free radicals during the chemotherapy treatment can be repressed or degraded to minimum by the help of antioxidants and may maintain a protection from the toxic side effects at the same time.