Effect of melatonin on steroidogenesis-related enzymes expression and testosterone synthesis following CoCl۲-induced hypoxia in TM۳ Leydig cells

Objective(s): This study examined the effects of melatonin treatment on steroidogenesis dysfunction and testosterone impairment, following CoCl۲-induced hypoxia in TM۳ Leydig cells. Materials and Methods: The TM۳ cells were divided into four groups. The first group received no treatment. The MLT group was treated with a concentration of ۱ mM melatonin. In the CoCl۲ group, ۰.۲ mM CoCl۲ was added to the medium to induce Hif۱α overexpression. The MLT+CoCl۲ group received ۰.۲ mM CoCl۲ and ۱ mM melatonin. After ۲۴ hr treatment, the cells and supernatants were collected and used for further determination. The MTT assay was performed to estimate the decrease in cell viability throughout the CoCl۲ and melatonin treatment. The mRNA and the protein levels were evaluated using Real-time PCR and Western blot analysis. The ELISA assay kit was used to detect the testosterone content.Results: CoCl۲ treatment caused Hif۱α overexpression in TM۳ Leydig cells. Moreover, CoCl۲ treatment of these cells led to considerable downregulation of Star, Hsd۳b۱, and Gata۴ well as Mtnr۱a and Mtnr۱b mRNA/protein expression coupled with testosterone content repression in the cell culture medium. Melatonin administration in CoCl۲ -treated cells decreased Hif۱α mRNA/protein expression, but had no significant effect on Star, Hsd۳b۱, Gata۴, Mtnr۱a mRNA/protein expression, and the testosterone level in the cell culture medium. Melatonin caused recovery of decrease in the Mtnr۱b gene and protein expression. Conclusion: There was no significant effect on steroidogenesis-related genes, proteins, and testosterone synthesis in the absence of gonadotropin treatment plus melatonin following CoCl۲-induced hypoxia in TM۳ Leydig cells.

کلیدواژه ها:

Hypoxia-inducible factor ۱- alpha subunit ، Leydig cells ، Melatonin ، Steroids ، Testosterone