Effects of Tomato and Olive Bioactive Compounds on tail regeneration in zebrafish larvae

In the present study, we used the zebrafish larvae as a tractable system to study the recruitment of neutrophils to the wound after caudal fin amputation and investigate the effects of Tomato and Olive Bioactive Compounds (TOBC) that promotes a regeneration permissive environment. TOBC derived from a food supplement registered by the Italian Health Ministry (patent No. EP2851080A1) based on spray-dried whole tomatoes and olive mill wastewater. The patented food supplement has been kindly provided by Janus Pharma (Rome, Italy). We first check for the presence of the compounds of interest and we measure through GC-MS the solution extract and we found the presence of tocopherol, lycopene, tyrosol, hydroxytyrosol and pinoresinol. Previous cellular studies have shown that TOBC exerts significant chemoprotective activity against the cellular insult of β-HCH, an environmental pollutant belonging to the OCPs family. Indeed, TOBC counteracts the cellular activities typical of β-HCH-induced carcinogenesis, such as activation of STAT3 and AhR, disruption of AR signalling, anti-apoptotic and proliferative activity, and increased ROS production and DNA damage. We used a transgenic zebrafish line – Tg(mpx:GFP) in which neutrophils are labelled by green fluorescent protein (GFP) – to screen how TOBC may affect neutrophil migratory behavior. To induce an acute inflammatory response the amputation was performed on 3 days post-fertilization larvae and before the amputation, each larvae was transferred to a beaker glass with 500 mL of clean and dechlorinated water and was anesthetized with 0.04% tricaine Pharmaq. We used TOBC solution at two concentration 0.5mg/L and 1mg/L on our model of tail transection to assess the possible effects of these compounds to stimulate tail regeneration through neutrophils recruitment. Moreover, we screen also the expression levels of two key proinflammatory cytokines such as TNFalpha and IL1beta. Our results show that the TOBC is able to promote a fast recruitment of neutrophils on fin resection and positive modulation of as TNFalpha and IL1beta, indicating a positive effect of these compounds on inflammatory modulation and fin regeneration. Our results are important to promote zebrafish model for model disease used in many pharmacology areas.