Abstract
The aim of this study was to evaluate the dose- and time-dependent in vitroeffects of isoquercitrin (ISO), a natural flavonoid with numerous biological properties on bovine spermatozoa during three different time periods (0 h, 2 h, 24 h). Bovine semen samples were diluted and cultivated in physiological saline solution containing 0.5% DMSO together with 200, 100, 50, 10, 5 and 1 μmol/L ISO. Spermatozoa motility was measured using the HTM IVOS CASA (Computer Assisted Semen Analyzer) system. The viability of spermatozoa was assessed by the metabolic (MTT) assay, production of superoxide radicals was quantified using the nitroblue tetrazolium (NBT) test, and chemiluminescence was used to evaluate the generation of reactive oxygen species (ROS). The results of the movement activity showed a significant increase in the motility during long term cultivation in case of concentrationsranging between 5 and 50 μmol/L ISO (P<0.05; 24 h). At the same time, supplementation of several concentrations of ISO led to a significant preservation of the cell viability (P<0.05 in the case of 50 μmol/L, P<0.01 with respect to 1 and 5 μmol/L, and P<0.001 in relation to 10 μmol/L; 24 h). ISO addition at 10 and 50 μmol/L also provided a significantly higher protection against superoxide (P<0.05) and ROS (P<0.001) overgeneration after a 24 h cultivation. We may suggest that supplementation of ISO to bovine spermatozoa, particularly at concentrations ranging between 10 and 50 μmol/L, may offer protection to the motility, viability and oxidative status of the sperm cells, particularly notable at 24 h.
Highlights
In recent years, oxidative damage that spermatozoa may suffer is receiving an ever-increasing interest
Moretti et al (2012) revealed in his study with human sperm, that quercetin, as well as rutin at concentration of 30 μmol/L were able to significantly (P
Reactive oxygen species are conventionally considered as detrimental by-products of cellular metabolism or exposure to xenobiotics, which generate a state of oxidative stress in susceptible cells
Summary
Oxidative damage that spermatozoa may suffer is receiving an ever-increasing interest. Excess oxygen during in vitro storage results in lipid peroxidation, causing membrane damage and a reduced sperm motility, viability and subsequent subfertility. Extensive effort has been invested to develop liquid storage procedures to hold spermatozoa for 24 h or longer, fertility is generally lower when insemination is performed with semen stored for more than 6 h (Donoghue and Donoghue, 1997). A successful liquid storage of spermatozoa is dependent on the maintenance of sperm metabolic activity. This is achieved by providing effective environmental conditions for the sperm survival, leading to the evolution of extenders that maintain the membrane integrity, motility, and fertilizing ability
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