Apoptotic and necrotic effects of plant extracts belonging to the genus Alchemilla L. species on HeLa cells in vitro

Abstract

Apoptotic and necrotic effects of plant extracts belonging to the genus Alchemilla on HeLa cells were investigated. The ratio of viable cells was determined by cell counter. Double staining method for the determination of apoptotic and necrotic index was carried out. The lowest apoptotic effect was found in Alchemilla oriturcica with the ratio of around 14% and the highest apoptotic effect found in Alchemilla trabzonica was 24%. Apoptotic effect was additionally determined by caspase-3 immunostaining. Increasing the concentrations of the extracts, especially in the level of 150 to 200 mg/mL caused the increase of necrotic effect on cancer cells. Particularly, the effects of A. oriturcica and A. trabzonica extracts with the density of 200 mg/mL were very high compared to the other species. The flavonoid components were detected in extracts of the species used for this study. The flavonoids identified are orientin (luteolin-8-C-glucoside), hyperoside (quercetin-3-O-galactoside) and isoquercetin (quercetin-3-glucoside) in Alchemilla erythropoda; rutin (quercetin-3-rutinoside), orientin, vitexin (apigenin-8-C-glucoside), hyperoside in Alchemilla ikizdereensis; rutin, hyperoside, isoquercetin (quercetin-3-glucoside) in A. oriturcica and hyperoside, isoquercetin and quercitrin (quercetin-3-O-rhamnoside) in A. trabzonica. Key words: Alchemilla, extract, HeLa cell, cytotoxicity, apoptosis, necrosis.