One aspect of modern research focused on prevention and cure of cancer is the investigation of new material of plant origin, with the objective of discovering new botanicals to use as active principle or to draw inspiration in the design of new molecular models1. For this aim, decoction (belonging to Ayurveda culture) and hydro-alcoholic extract (closer to occidental tradition) of the Hemidesmus indicus R.Br. (Asclepiadaceae) root was studied . The phytochemical characterisation of the hydro-alcoholic extract, performed by RP-HPLC-DAD, showed an higher quantity of vanillin isomers and derivatives (2-hydroxy-4-methoxybenzaldehyde, 3-hydroxy-4-methoxybenzaldehyde and 2-hydroxy-4-methoxybenzoic acid) in comparison to decoction. In the aqueous preparation, in addition to these molecules, lupeol, lupeol acetate, β-amyrin acetate and β-sitosterol were identified by GC-MS. Being aware that oxidative stress is a key factor in the cancer development, the study of bio-activities started from the evaluation of the antioxidant capacity through spectrophotometric methods (DPPH, ABTS and β-carotene bleaching test) and HPTLC-bioautographic assays4 (DPPH, ABTS test). At a later stage, the evaluation of geno-protective (SOS-Chromotest) and anti-proliferative activity (using CCRF-CEM, CEM/ADR5000, MCF7, A549, MDA-MB-231, LoVo, HepG2, K562 and Jurkat cell lines) was carried out, focusing on the vanillin derivatives identified and quantified in H. indicus. Plant decoction had already evidenced an important anti-leukemic effect through the modulation of different critical targets2,3, and the vanillin derivative showed relevant data for the geno-protective and anti-proliferative activity. In particular, the most active vanillin isomer 2-hydroxy-4-methoxybenzaldehyde showed IC50 values of 79.52 ± 18.27 μM against K562, 85.39±1.70 μM against CCRF-CEM and 86.46 ± 3.66 μM against Jurkat. Further in-depth analysis regarding anti-proliferative bioactivity of traditional preparations, identification and quantification of other characteristic compounds and more selective extractions are still in progress. In conclusion: H. indicus evidenced promising data against Jurkat (63.79±7.97 μg/mL), CCRF-CEM (46.23±1.12 μg/mL), Hep-G2 (34.50±0.14 μg/mL) and LoVo (29.84±0.24 μg/mL); H. indicus hydro-alcoholic extract and decoction were more effective than 2-hydroxy-4-methoxybenzaldehyde against Hep-G2 cell, pointing out possible synergistic (agonistic) activity of minor compounds.
In vitro evaluation of anti-proliferative and geno-protective activity of Hemidesmus indicus crude drug extracts
TACCHINI, Massimo;SPAGNOLETTI, Antonella;BROGNARA, Eleonora;GAMBARI, Roberto;GUERRINI, Alessandra;SACCHETTI, Gianni
2014
Abstract
One aspect of modern research focused on prevention and cure of cancer is the investigation of new material of plant origin, with the objective of discovering new botanicals to use as active principle or to draw inspiration in the design of new molecular models1. For this aim, decoction (belonging to Ayurveda culture) and hydro-alcoholic extract (closer to occidental tradition) of the Hemidesmus indicus R.Br. (Asclepiadaceae) root was studied . The phytochemical characterisation of the hydro-alcoholic extract, performed by RP-HPLC-DAD, showed an higher quantity of vanillin isomers and derivatives (2-hydroxy-4-methoxybenzaldehyde, 3-hydroxy-4-methoxybenzaldehyde and 2-hydroxy-4-methoxybenzoic acid) in comparison to decoction. In the aqueous preparation, in addition to these molecules, lupeol, lupeol acetate, β-amyrin acetate and β-sitosterol were identified by GC-MS. Being aware that oxidative stress is a key factor in the cancer development, the study of bio-activities started from the evaluation of the antioxidant capacity through spectrophotometric methods (DPPH, ABTS and β-carotene bleaching test) and HPTLC-bioautographic assays4 (DPPH, ABTS test). At a later stage, the evaluation of geno-protective (SOS-Chromotest) and anti-proliferative activity (using CCRF-CEM, CEM/ADR5000, MCF7, A549, MDA-MB-231, LoVo, HepG2, K562 and Jurkat cell lines) was carried out, focusing on the vanillin derivatives identified and quantified in H. indicus. Plant decoction had already evidenced an important anti-leukemic effect through the modulation of different critical targets2,3, and the vanillin derivative showed relevant data for the geno-protective and anti-proliferative activity. In particular, the most active vanillin isomer 2-hydroxy-4-methoxybenzaldehyde showed IC50 values of 79.52 ± 18.27 μM against K562, 85.39±1.70 μM against CCRF-CEM and 86.46 ± 3.66 μM against Jurkat. Further in-depth analysis regarding anti-proliferative bioactivity of traditional preparations, identification and quantification of other characteristic compounds and more selective extractions are still in progress. In conclusion: H. indicus evidenced promising data against Jurkat (63.79±7.97 μg/mL), CCRF-CEM (46.23±1.12 μg/mL), Hep-G2 (34.50±0.14 μg/mL) and LoVo (29.84±0.24 μg/mL); H. indicus hydro-alcoholic extract and decoction were more effective than 2-hydroxy-4-methoxybenzaldehyde against Hep-G2 cell, pointing out possible synergistic (agonistic) activity of minor compounds.I documenti in SFERA sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
