Effects of hydroethanolic extracts of Terminalia chebula and Thymbra spicata on ram fresh semen under normal and oxidative stress conditions.

The objective of this study was to evaluate protective effects of hydroethanolic extracts of Terminalia chebula and Thymbra spicata on viability, lipid peroxidation (LPO) and DNA integrity of ram fresh semen under normal and oxidative stress (OS) conditions. Antioxidant activities of different concentrations of Terminalia chebula and Thymbra spicata extracts were evaluated with DPPH assay. Semen samples were taken from three fertile adult rams. After diluting semen with Tris-base extender, different concentrations of Terminalia chebula and Thymbra spicata (30, 300, and 3000 μg/ml) extracts were used under normal and induced OS conditions. The group not receiving any supplements was considered as control group. A total of 50 μM hydrogen peroxide was used to induce OS. MTT solution was added to each of treatment groups which were kept in an incubator at 37°C for 2 h. After incubation, readings were obtained by ELISA reader. DNA integrity and LPO were determined with acridine orange (AO) staining and malondialdehyde (MDA) assay. Higher concentrations of Terminalia chebula and Thymbra spicata extracts preserved viability and DNA integrity while reducing MDA concentrations compared to other treatment groups. Also, under induced OS, higher concentrations of both extracts reduced detrimental effects of H2 O2 . In conclusion, it seems that addition of Terminalia chebula and Thymbra spicata extracts can reduce induced OS in spermatozoa.