The impact of organic hydroperoxides and a red palm oil supplemented diet on spermatogenesis, sperm function and sperm apoptosis

Abstract

Many environmental, physiological, and genetic factors have been shown to impair sperm function through oxidative damage. Oxidative stress (OS) arises as a consequence of excessive reactive oxygen species (ROS) production and/or impaired antioxidant defence mechanisms. The decline in male reproductive health generated considerable public and scientific concerns about the possible role of environmental contaminants. A better understanding of how OS affects sperm function will be beneficial as it might help in the design of new and effective treatment strategies to combat the problem of increasing male subfertility. Studies have suggested that antioxidant nutrients and/or medicines play a protective role in human health. Crude red palm oil (RPO) is known to be the richest natural plant source of antioxidants such as carotenoids, tocopherols and metalloporpheryns. The aims of this study were twofold: (i) To establish an in vivo animal model of OS by exposing rat to organic hydroperoxide such as t-butyl hydroperoxide (tbHP) and cumene hydroperoxide (cHP) through repeated intraperitoneal injections that can be used for studying these effects on testicular tissue, epididymal sperm and sperm function as well as male reproductive parameters in general. (ii) To investigate the effects of a RPO supplemented diet on male reproductive parameters and tissue in animals exposed to OS. In the first part of the study, male Wistar rats aged 10-12 weeks were randomly placed in groups and received standard rat chow (SRC) and water ad lib. Animals were injected intraperitoneally with saline (0.5 ml), t-butyl hydroperoxide (5µM, 10µM, 20µM and 40µM; 0.5 ml) or cumene hydroperoxide cHP (2.5µM, 5µM, 10µM and 20µM; 0.5 ml) over a 60 day period. In the second part, male Wistar rats aged 10-12 weeks were placed randomly in three groups and fed with SRC. Group 1 received no supplement while the food of groups 2 and 3 were supplemented with 2 mL and 4 mL RPO (in 25 gm SRC/day) respectively. Each group was further divided into 3 subgroups and injected intraperitoneally daily with either saline, 10µM cHP or 20µM tbHP respectively. This was done for 5 consecutive days per week over a 60 day period. Sperm concentrations, and motility, lipid peroxidation, superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) activities as well as apoptosis were assessed.