ORIGINAL ARTICLE In vitro toxicity and genotoxic activity of aqueous leaf and fruit extracts of Ruscus hypophyllum L. Luc Verschaeve 1,2 • Hayet Edziri 3 • Roel Anthonissen 1 • Dalenda Boujnah 4 • Fethia Skhiri 5 • Mahjoub Aouni 3 • Maha Mastouri 3 Received: 4 November 2016 / Revised: 19 April 2017 / Accepted: 18 August 2017 Ó Franciszek Go´rski Institute of Plant Physiology, Polish Academy of Sciences, Krako´w 2017 Abstract Ruscus hypophyllum L. is a rare Mediterranean plant which is used in the traditional medicine. We studied its phenolic content and in vitro toxicity and genotoxicity using the neutral red uptake (NRU) test, the bacterial Vitotox test, and the comet assay in human C3A hepatic cells. Aqueous leaf and fruit extracts were investigated. Antigenotoxicity against 4-nitroquinoline-oxide (4NQO, 0.4 lg/mL) and Benzo(a)pyrene (BaP, 800 lg/mL) was also investigated with the Vitotox test. The extracts appeared to be genotoxic only at high exposure levels in the comet assay. There was no indication of a genotoxic activity in the Vitotox test and also no indication of antigenotoxicity. The moderate polyphenol content may provide an explanation for the absence of antigenotoxicity. Keywords Ruscus hypophyllum L.  Phenolic content  Genotoxicity  Antigenotoxicity  Vitotox test  Comet assay Introduction Medicinal plants have been used throughout human his- tory. They have the ability to synthesize a wide variety of chemical compounds that may be used to perform or modify important biological functions. Many extracts prepared from plants have biological activities in vitro and in vivo. In recent years, there has been great interest in investigating compounds from plants and their effects on DNA. This is because preparations from the traditional medicinal plants were seldom tested the way our modern pharmaceutical compounds are, and hence, they may potentially have unknown adverse effects. Such effects may be mutation induction and carcinogenicity. Com- pounds from plant origin can be mutagenic but also antimutagenic, and hence, they can contribute to as well increases as reductions in the incidence of cancer in the population (Ames 1983). It may, therefore, be interesting to investigate not only genotoxicity but also potential antigenotoxic properties of plant extracts with alleged medicinal properties. Ruscus hypophyllum L. belongs to the Liliacea family. It is a rare and endangered species which is especially found in the Northern region of Tunisia. It is native in N. Africa, as far east as Tunisia, but extends to the northern littoral of the Mediterranean (Yeo 1968). It is a perennial, dioecious, evergreen rhizomatous geophyte. The mature plant shrub may reach about 46 cm in height. It has a creeping rootstock and leaf-like phylloclades. True leaves are small green appen- dages around the flowers. The bracts are located in the middle Communicated by MJ Reigosa. & Luc Verschaeve Luc.Verschaeve@wiv-isp.be Hayet Edziri jaziri_hayet@yahoo.fr 1 Toxicology Unit, Scientific Institute of Public Health, Brussels, Belgium 2 Department of Biomedical Sciences, University of Antwerp, Antwerp, Belgium 3 Laboratory of Transmissible Diseases and Biologically Active Substances, Faculty of Pharmacy, University of Monastir, Monastir, Tunisia 4 Olive Tree Institute, Sousse Station, Sousse, Tunisia 5 Laboratory of Genetic Biodiversity and Valorisation of Bioresources, Higher Institute of Biotechnology of Monastir, Monastir, Tunisia 123 Acta Physiol Plant (2017) 39:206 DOI 10.1007/s11738-017-2505-7