Reproductive Toxicology 31 (2011) 239–246 Contents lists available at ScienceDirect Reproductive Toxicology journal homepage: www.elsevier.com/locate/reprotox Effect of trans-resveratrol on induced oxidative stress in human sperm and in rat germinal cells G. Collodel a,b , M.G. Federico a , M. Geminiani a , S. Martini c,e , C. Bonechi c,e , C. Rossi c,e , N. Figura d , E. Moretti a,b,∗ a Department Biomedical Sciences, Applied Biology Section, Italy b Interdepartmental Centre for Research and Therapy of Male Infertility, Italy c Department of Pharmaceutical and Applied Chemistry, University of Siena, Italy d Department of Internal Medicine, Endocrine-Metabolic Sciences and Biochemistry, University of Siena, Italy e Centre for Colloid and Surface Science (CSGI), University of Florence, Via della Lastruccia, 3, Sesto Fiorentino, FI, Italy article info Article history: Received 24 September 2010 Received in revised form 2 November 2010 Accepted 19 November 2010 Available online 30 November 2010 Keywords: Human sperm Oxidative stress Rat germ cells STAPUT TEM Trans-resveratrol abstract Resveratrol is a phytoalexin with antioxidant properties. We evaluated resveratrol toxicity in swim- up selected human sperm and in rat spermatocytes and spermatids separated by the STAPUT method. Resveratrol antioxidant activity was tested against lipid peroxidation (LPO) induced by tert-butyl hydroperoxide. LPO was evaluated using the C11-BODIPY 581/591 probe and transmission electron microscopy in samples incubated with and without resveratrol. LD50 for human sperm and rat spermatids was 50 M; spermatocytes were more sensitive to resveratrol cytotoxicity. Sperm motility increased pro- gressively at 30 M, 15 M and 6 M. 15 M resveratrol acts against LPO, preserving sperm chromatin and plasma membranes. LPO were more marked in spermatocytes than in spermatids and the effect of resveratrol was more evident in spermatocytes. In this study, the scavenger properties of resvera- trol were demonstrated in vitro in human sperm and rat germ cells, thus resveratrol could be added to the media used in assisted reproduction techniques and cryopreservation when oxidative stress is exacerbated. © 2010 Elsevier Inc. All rights reserved. 1. Introduction Reactive oxygen species (ROS) are products of normal cellular metabolism and they play a functional role in many cell types as second messengers. When ROS are produced at a very low concen- tration, they trigger cell signaling events and regulate physiological function [1,2]. Sperm were the first type of cell reported to produce free radicals and MacLeod [3] noted that the incubation of sperm under high oxygen tension leads to a rapid loss of sperm motility. Low level production of ROS by sperm supports some main functions, such as capacitation, acrosome reaction, zona pellucida binding and oocyte fusion [4]. Nevertheless, uncontrolled ROS production can play an important role in causing sperm aberration, leading to infertility. What is usually called “oxidative stress” appears to be the result of defects in the balance between the concentration of ROS and the antioxidant scavenging system. Sperm membranes are rich in ∗ Corresponding author at: Department of Biomedical Sciences, Applied Biology Section, University of Siena, Policlinico Santa Maria alle Scotte, Viale Bracci, 53100 Siena, Italy. Tel.: +39 577 233511; fax: +39 0577 233527. E-mail address: moretti@unisi.it (E. Moretti). polyunsaturated fatty acid, which makes them very susceptible to oxygen-induced damage mediated by lipid peroxidation (LPO) [5]. Oxidative stress has been suggested to be an important factor in the aetiology of poor sperm function through peroxidative damage to the cell membrane, to DNA (inducing single- and double-strand DNA breaks) and to proteins [6–8]. Seminal plasma and sperm are endowed with an array of pro- tective antioxidants, such as the glutathione peroxidase/reductase system, superoxide dismutase, catalase and low-molecular weight antioxidants, vitamin E, vitamin C, urate, and albumin, which scav- enge ROS in order to prevent possible cellular damage [9]. One of the rational strategies for counteracting oxidative stress is to increase the scavenging capacity of seminal plasma. However, the use of antioxidant supplementation to reverse the effect of ROS is still being debated [2,10]. Resveratrol (RSV) is a natural phytoalexin with antioxidant properties that is widely consumed in the Mediterranean diet in the form of peanuts, grapes and wine. Interest in the compounds present in wine has increased since epidemiological studies indi- cated an inverse correlation between red wine consumption and the incidence of cardiovascular disease [11]. In addition, RSV has exhibited a broad range of biological activities, including antiin- flammatory, antiviral and antitumoral properties [12]. 0890-6238/$ – see front matter © 2010 Elsevier Inc. All rights reserved. doi:10.1016/j.reprotox.2010.11.010