In vitro Assessments of Cytotoxic and Cytostatic Effects of Asparagus aphyllus, Crataegus aronia, and Ephedra alata in Monocultures and Co-Cultures of Hepg2 and THP-1-Derived Macrophages

Introduction: Herbal-based medicines are widely used for the prevention and treatments of diverse diseases especially in growing countries as well as many developed countries. Although some of herbal-based medicines have promising therapeutic properties, many of them remain untested and their safety and efficacy were not scientifically assessed. Based on knowledge from traditional Greco-Arab herbal medicine, this in vitro study aims to evaluate cytotoxic and cytostatic effects of three traditionally used anti-diabetic and anti-cancer medicinal plants. Out of the traditional medicinal plants, Asparagus aphyllus, Crataegus aronia, and Ephedra alata are widely used in many Mediterranean countries as natural remedies. Methods: Human THP-1- derived macrophages, HepG2 cells and their co-cultures were used in this in vitro study. Cells were treated for 24h (cytotoxic effects) and 72h (cytostatic effects) with increasing concentrations (0-1000 μg/ml) of water/ethanol extracts from Asparagus aphyllous (AA-extract), Crataegus aronia (CA-extract) and Ephedra alata (EA-extract). Cytotoxic and cytostatic effects were assessed using MTT assay and LDH assay. Results: No significant cytotoxic effects were seen with the three extracts up to concentration of 500 μg/ml. A slight cytotoxic effect was observed with CA-extractin HepG2 monocultures at concentrations higher than 500 μg/ml. Significant cytostatic effects were measured with CA-extract and EA-extract in monocultures and cocultures. The cytostatic activity of the extracts was more potent in co-cultures reaching IC50 of 178 μg/mL and 380 μg/mL for CA-extract and EA-extract, respectively. Conclusion: These results indicate that the traditionally known anti-cancer effects of CA-extract and EA-extract might be mediated in part through cytostatic effects.