In vitro effect of platelet-rich plasma (PRP) on morpho-functional characteristics, oxidative stress, apoptosis and CatSper-1 ion channel in short-term stored (5 °C) ram semen

Abstract

The objective of this study was to examine the impact of platelet-rich plasma (PRP) on the morpho-functional characteristics, oxidative stress, apoptosis and CatSper-1 ion channel in short-term stored ram semen at 5 °C. The study was conducted in six independent replicates on pooled semen samples collected by artificial vagina from six sexually active Akkaraman rams. The pooled semen samples were diluted to a concentration of 400 million spermatozoa per milliliter with a tris-based diluent including 6 PRP concentration (1, 2, 4, 8, 10, 20 %) and no PRP (control) at 38 °C. Following the reduction of the temperature to 5 °C, diluted semen samples were stored for a period of 96 hours. Motility (total, progressive, rapid, medium, low), kinematic (VCL, VSL, VAP, LIN, STR, WOB, ALH, BCF), morphological (plasma membrane integrity, abnormality), flow-cytometric (mitochondrial activity, acrosomal damage, viability, apoptosis), oxidative stress (MDA, GSH, GSH-Px, CAT), and CatSper-1 analyses were performed at both hours zero (initial measurement) and 96. In all groups, a time-dependent deterioration was observed in nearly all parameters at 96 h in comparison to the 0 h. Compared to the control, live spermatozoon rates (8 % and 10 %), GSH-Px (4 %, 8 % and 10 %) and CAT (4 % and 8 %) activities, and CatSper-1 levels at 96 h were higher, while the apoptotic spermatozoon rates (8 % and 10 %) and MDA levels (4 %, 8 % and 10 %) were lower in semen samples containing PRP. However, in semen samples containing 20 % PRP, the apoptotic spermatozoon rates and MDA levels at 96 h were higher, while GSH-Px activity and CatSper-1 levels were lower than in the control. As a result, it was concluded that the addition of 4–10 % PRP to the diluent containing tris+egg yolk has an effective role in preventing oxidative, apoptotic and ion channel (CatSper-1) damages in ram spermatozoa stored for a short time at 5 °C, while the concentration of 20 % is harmful.