Abstract: This study aimed to assess the in vitro effects of different concentrations of the Viscum album extract on the motility, viability, and reactive oxygen species (ROS) production by rabbit spermatozoa during different time periods (0, 2, and 8h). Spermatozoa motility was assessed by using the CASA (Computer aided sperm analysis) system. Cell viability was evaluated by using the metabolic activity MTT assay, and the luminol-based luminometry was applied to quantify the ROS formation. The CASA analysis revealed that low Viscum concentrations were able to prevent a rapid decline of spermatozoa motility, especially in the case of concentrations ranging between 1 and 5 µg/mL (P
Abstract: In vitro storage and processing of animal semen represents a risk factor to spermatozoa vitality, potentially leading to reduced fertility. A variety of substances isolated from natural sources may exhibit protective or antioxidant properties on the spermatozoon, thus extending the lifespan of stored ejaculates. This study compared the ability of different concentrations of the Salvia officinalis extract on the motility, mitochondrial activity, viability and reactive oxygen species (ROS) production by bovine spermatozoa during different time periods (0, 2, 6 and 24 h) of in vitro culture. Spermatozoa motility was assessed using the Computer-assisted sperm analysis (CASA) system. Cell viability was examined using the metabolic activity MTT assay, the eosin-nigrosin staining technique was used to evaluate the sperm viability and ROS generation was quantified using luminometry. The CASA analysis revealed that the motility in the experimental groups supplemented with 0.5-2 µg/mL Salvia extract was significantly lower in comparison with the control (P
Abstract: The present in vitro study was designed to reveal whether amygdalin (AMG) is able to cause changes to the motility, viability and mitochondrial activity of rabbit spermatozoa. New Zealand White rabbits (n = 10) aged four months were used in the study. Semen samples were collected from each animal and used for the in vitro incubation. The samples were divided into five equal parts and diluted with saline supplemented with 0, 0.5, 1, 2.5 and 5 mg/mL AMG. At times 0h, 3h and 5h spermatozoa motion parameters were assessed using the SpermVision™ computer-aided sperm analysis (CASA) system, cell viability was examined with the metabolic activity (MTT) assay, and the eosin-nigrosin staining technique was used to evaluate the viability of rabbit spermatozoa. All AMG concentrations exhibited stimulating effects on the spermatozoa activity, as shown by a significant preservation of the motility (P
Abstract: The potential neuroprotective effect of Phyllantus
nuriri against Fe2+ and sodium nitroprusside (SNP) induced oxidative
stress in mitochondria of rats brain was evaluated. Cellular viability
was assessed by MTT reduction, reactive oxygen species (ROS)
generation was measured using the probe 2,7-dichlorofluoresce
indiacetate (DCFH-DA). Glutathione content was measured using
dithionitrobenzoic acid (DTNB). Fe2+ (10μM) and SNP (5μM)
significantly decreased mitochondrial activity, assessed by MTT
reduction assay, in a dose-dependent manner, this occurred in parallel
with increased glutathione oxidation, ROS production and lipid
peroxidation end-products (thiobarbituric acid reactive substances,
TBARS). The co-incubation with methanolic extract of Phyllantus
nuriri (10-200 μg/ml) reduced the disruption of mitochondrial
activity, gluthathione oxidation, ROS production as well as the
increase in TBARS levels caused by both Fe2+ and SNP in a dose
dependent manner. HPLC analysis of the extract revealed the
presence of gallic acid (20.540.01), caffeic acid (7.930.02), rutin
(25.310.05), quercetin (31.280.03) and kaemferol (14.360.01).
This result suggests that these phytochemicals account for the
protective actions of P. niruri against Fe2+ and SNP -induced
oxidative stress. Our results show that P. nuriri consist important
bioactive molecules in the search for an improved therapy against the
deleterious effects of Fe2+, an intrinsic producer of reactive oxygen
species (ROS), that leads to neuronal oxidative stress and
neurodegeneration.