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ETHNOPHARMACOLOGICAL RELEVANCE: Tibetan medicine get used to use the flowers of Gentiana straminea Maxim. to cure inflammation of stomach and intestines, hepatitis, cholecystitis, etc. The flowers of Gentiana macrophylla Pall. have been traditionally treated as an anti-inflammatory agent to clear heat in Mongolian medicine. In traditional Chinese medicine, Gentiana macrophylla Pall. and Gentiana straminea Maxim. have also been used under the name "Gentianae Macrophyllae Radix" and prescribed for the treatment of pain and inflammatory conditions.AIM OF STUDY: The present study evaluated the pharmacological effects of two species of "Radix Gentianae Macrophyllae" in experimental inflammation and pain models, and determined the chemical compounds that may correlate with their pharmacological activities. The comparison is needed to identify whether the two related plants can be used interchangeably.
MATERIALS AND METHODS: We evaluated the pharmacological effects of the flowers of Gentiana macrophylla Pall. and Gentiana straminea Maxim. in experimental inflammation and pain models. An HPLC-MS method was developed to analyze the chemical composition. The effects of Gentiana macrophylla Pall. and Gentiana straminea Maxim. on the p65 and p50 phosphorylation were examined by immunblotting. NF-κB transcriptional activity was measured using the luciferase assay, in vitro kinase assay and Griess reaction.
RESULTS: The extracts of Gentiana macrophylla Pall. and Gentiana straminea Maxim. possessed significant antinociceptive and anti-inflammatory activities. Flavonoids, secoiridoid glycosides and triterpines were determined in the extracts and may be the basis of the observed pharmacological effects. Nuclear translocation of p65, p50 and NF-κB transcriptional activity induced by LPS were suppressed by Gentiana macrophylla Pall. and Gentiana straminea Maxim.
CONCLUSION: The results clearly demonstrated that the chemical composition and pharmacological activities of the two herbs were similar, which support the interchangeability among the two herbs when using them in folk medicine.
A rapid method combining microwave-assisted extraction (MAE) and high-speed counter-current chromatography (HSCCC) was applied for preparative separation of six bioactive compounds including loganic acid (I), isoorientin-4'-O-glucoside (II), 6'-O-β-d-glucopyranosyl gentiopicroside (III), swertiamarin (IV), gentiopicroside (V), sweroside (VI) from traditional Tibetan medicine Gentiana crassicaulis Duthie ex Burk. MAE parameters were predicted by central composite design response surface methodology. That is, 5.0 g dried roots of G. crassicaulis were extracted with 50 mL 57.5% aqueous ethanol under 630 W for 3.39 min. The extract (gentian total glycosides) was separated by HSCCC with n-butanol/ethyl acetate/methanol/1% acetic acid water (7.5:0.5:0.5:3.5, v/v/v/v) using upper phase mobile in tail-to-head elution mode. 16.3, 8.8, 12., 25.1, 40.7, and 21.8 mg of compounds I-VI were obtained with high purities in one run from 500 mg of original sample. The purities and identities of separated components were confirmed using HPLC with photo diode array detection and quadrupole TOF-MS and NMR spectroscopy. The study reveals that response surface methodology is convenient and highly predictive for optimizing extraction process, MAE coupled with HSCCC could be an expeditious method for extraction and separation of phytochemicals from ethnomedicine.