Effect of vitamin E in the culture medium on the expression of apoptotic genes, the level of antioxidants and sex hormones in ovarian tissue treated with cyclophosphamide

Abstract

One of the most important challenges in the field of health is maintaining fertility in young women in cancer patients. Premature ovarian failure and ultimately infertility is the consequence of using chemotherapy drugs for these patients. Cyclophosphamide, which is one of the most widely used drugs in the field of chemotherapy, has been repeatedly investigated for its adverse effects on the reproductive system of women. Assisted reproductive techniques such as in vitro fertilization and freezing of ovaries or eggs and maturation and development of ovarian follicles are among the most important strategies for maintaining fertility in women who have used the drug cyclophosphamide. Considering the possible mechanisms of the effects of this drug such as conditions of oxidative stress and Apoptosis The use of antioxidants such as vitamin E or alphatocopherol can improve the success rate of follicular maturation and development in assisted reproductive techniques of patients who have received cyclophosphamide, so in this study, we investigated the effect of added vitamin E in the culture medium of antral follicles of mice that underwent are treated with cyclophosphamide, we examine the parameters of oxidative stress and hormone levels and the expression of genes related to apoptosis. Method: 6-8 weeks old adult female mice were divided into two main control groups and cyclophosphamide treatment group, which received a dose of 150 mg/kg intraperitoneally, on the fifth day of treatment, they receiving cyclophosphamide to isolate and culture antral follicles by moving the cervical vertebrae. Follicles were cultured in the culture medium containing and lacking vitamin E for 4 days, and then on the fourth day, by preparing the cell supernatant, the levels of estradiol and progesterone hormones, SOD, GPX, TAC, and MDA indicators were measured using the ELISA technique and protein gene expression levels. BAX and bcl-2 were evaluated by PCR. Results: The results of this research showed that the amount of antioxidant indicators SOD, GPX, TAC and MDA in the group that received only cyclophosphamide drug significantly decreased and increased respectively (P<0.01), while compared to the group treated with cyclophosphamide. were treated and their culture medium was enriched with vitamin E, these indicators increased and decreased respectively. The amount of estradiol hormone in the treatment group where vitamin E was added to the culture medium showed a significant increase compared to the empty treatment group (P<0.01). While the amount of progesterone hormone did not show any significant difference in all groups. BAX apoptotic gene expression in the cyclophosphamide treatment group showed a significant increase compared to the treatment group that was enriched with vitamin E in the culture medium (P<0.01). The expression level of anti-apoptotic bcl-2 gene in the group treated with cyclophosphamide decreased compared to the group treated with vitamin E (P<0.01).