Apoptosis Induction byPolygonum minusIs Related to Antioxidant Capacity, Alterations in Expression of Apoptotic-Related Genes, and S-Phase Cell Cycle Arrest in HepG2 Cell Line

Abstract

Polygonum minus (Polygonaceae) is a medicinal herb distributed throughout eastern Asia. The present study investigated antiproliferative effect of P. minus and its possible mechanisms. Four extracts (petroleum ether, methanol, ethyl acetate, and water) were prepared by cold maceration. Extracts were subjected to phytochemical screening, antioxidant, and antiproliferative assays; the most bioactive was fractionated using vacuum liquid chromatography into seven fractions (F1–F7). Antioxidant activity was measured via total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing antioxidant power (FRAP) assays. Antiproliferative activity was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Most active fraction was tested for apoptosis induction and cell cycle arrest in HepG2 cells using flow cytometry and confocal microscopy. Apoptotic-related gene expression was studied by RT-PCR. Ethyl acetate extract was bioactive in initial assays. Its fraction, F7, exhibited highest antioxidant capacity (TPC; 113.16 ± 6.2 mg GAE/g extract, DPPH; EC50: 30.5 ± 3.2 μg/mL, FRAP; 1169 ± 20.3 μmol Fe (II)/mg extract) and selective antiproliferative effect (IC50: 25.75 ± 1.5 μg/mL). F7 induced apoptosis in concentration- and time-dependent manner and caused cell cycle arrest at S-phase. Upregulation of proapoptotic genes (Bax, p53, and caspase-3) and downregulation of antiapoptotic gene, Bcl-2, were observed. In conclusion, F7 was antiproliferative to HepG2 cells by inducing apoptosis, cell cycle arrest, and via antioxidative effects.

Anti-proliferative,in vitroantioxidant, and cellular antioxidant activities of the leaf extracts fromPolygonum minusHuds: Effects of solvent polarity

“…Based on such a classification, PM-MeOH and PM-W are considered to have extremely high antioxidant power. The respective FRAP values for PM-MeOH and PM-W extracts were about two and thirteen times higher than the values that were previously reported: 781.32 ± 4.2 µmol Fe 2+ /g MeOH extract [24] and 68.3 ± 3.3 µmol Fe 2+ /g water extract. [21] ABTS •+ radical-scavenging activity ABTS •+ reagent is a stable radical monocation that is directly produced from the oxidation of ABTS by potassium persulfate.…”

“…PM-MeOH and PM-W scavenging activities appeared to increase rapidly and reached their maximum values at~500 μg/mL. The IC 50 values for PM-MeOH and PM-W extracts were 125.51 ± 4.70 and 210.16 ± 1.35 μg/mL, respectively, and these values imply a higher scavenging activity compared with that reported by Ghazali et al, [24] which were 540.0 ± 10.8 μg/mL for PM-MeOH and 472.5 ± 15.2 μg/mL for PM-W.…”

“…[23] The ethyl acetate (EA) fraction of P. minus was cytotoxic and capable of inducing apoptosis and S-phase cell-cycle arrest in HepG2 cells. [24] P. minus extracts have also been reported to show the following activities: antioxidant, [25,26] anti-HSV1, anti-vesicular stomatitis virus,- [27] anti-fungal, anti-ulcer, [28] anti-hyperlipidemic, [29] anti-acetylcholinesterase, [30] antimicrobial, and cytotoxicity towards HeLa cells. [27,31] Other members of Polygonaceae have also been reported to show antimicrobial, antioxidant, anticancer, cytotoxic, anti-angiogenic, and anticholinesterase activities.…”

Phytochemical and biological research of Polygoneae medicinal resources

“…Apoptosis induction by P. minus is related to antioxidant capacity, alterations in expression of apoptotic-related genes, and S-phase cell cycle arrest in HepG2 cell line (Mohd Ghazali et al, 2014). Resveratrol-4-O-D-(2 0 -galloyl)-glucopyranoside isolated from P. cuspidatum exhibits anti-hepatocellular carcinoma viability by inducing apoptosis via the JNK and ERK pathway .…”

Effect of Polygonum Minus (Knotweed) Leaves Extract on the Histopathological Changes of Kidney in Mice (Mus Musculus) Induced by Mercuric Chloride

“…Polygonum minus or kesum in Indonesia is traditionally used as food ingredient, hair dandruff, and also indigestion (Ghazali et al, 2014). It can be found also in Sumatera and Kalimantan.…”