Ethanol extract of Cimicifugae rhizoma exerted more potent anti- inflammatory and tumor suppressor activities compared with methanol and water extracts

Cimicifugae rhizoma (C. rhizoma) called Shengma (Chinese), Seungma (Korean), and Shoma (Japanese) is primarily derived from Cimicifuga heracleifolia Komarov or Cimicifuga foetida Linnaeus. C. rhizoma is used for anti-inflammatory, analgesic, antipyretic remedy and alternative for hormone replacement therapy. The aim of this study was to determine the dose-dependent anti-inflammatory and tumor suppressor activities using methanol, ethanol and water extract of C. rhizoma. Methanol, ethanol and water extracts of Cimicifuga heracleifolia Komarov were obtained. Mouse leukaemic monocyte macrophage cell line (RAW 264.7) was loaded in the presence of C. rhizoma at final concentrations that ranged from 10 to 50 μg/mL. The production of Nitric Oxide (NO) in lipopolysaccharides-induced RAW 264.7 cells was quantified. Cycloxygenase-2 (COX-2) mRNA expression was evaluated using Lipopolysaccharides (LPS)-stimulated RAW 264.7 cells with semiquantitative Reverse Transcription-Polymerase Chain Reaction (RT-PCR). Lung carcinoma cell lines (A549) were treated with C. rhizoma at final concentrations that ranged from 200 to 800 μg/mL and expressions of caspase-3 and Superoxide Dismutase-2 (SOD-2) were tested with RT-PCR. No changes of cell viability were noted after treatment with methanol, ethanol, and water extracts of C. rhizoma. Stimulation with LPS for 24 h led to a robust increase in the NO production, but C. rhizoma significantly suppressed NO by the LPS-stimulated RAW 264.7 cells in all groups. Pretreatment with absolute ethanol extract of C. rhizoma suppressed the LPS-stimulated COX-2 expression. The results showed that C. rhizoma extract significantly showed anticancer effects in methanol and ethanol groups. The expression of caspase-3 and SOD-2 increased with the increase of exposure time with C. rhizoma extract. Within the limits of this study, C. rhizoma showed anti-inflammatory effect using RAW 264.7 cell line and tumor suppressor activities on A549 cells. Absolute ethanol extract showed the highest antiinflammatory and tumor suppressor activities on these experimental settings.