Studies On The Chemical Components And Hematostatic Activity Of Traditional Tibetan Herb Lamiophlomis Rotate

The genus Lamiophlomis kudo (Labiatae) is represented by only one species, Lamiophlomis rotata (Benth.) Kudo, which is widely spread over high mountainous regions in Tibet and used in traditional Chinese medicine to stop bleeding, remove blood stasis, subdue swelling, alleviate pain and as antiseptic, anti-inflammation agents as well as immunity-enhancer. To study the chemical components and to find the hematostatic fraction of this plant, phytochemical investigation has been carried out under the guidance of pharmacological evaluation. Seventeen compounds, involving alkaloids, iridoid glycosides, flavonoids, phenethanol glycosides, coumarins, phenols, phenolic acids, were isolated from this plant and identified on the basis of spectroscopic analysis. Hematostatic activities of aqueous extract of the aerial parts of L. rotata, fractions of different polarity and several compounds were evaluated.Aqueous extract of the aerial parts was partitioned with EtOAC and n-BuOH successively to form three fractions of different polarity. EtOAC fraction was subjected to a normal phase SG crude separation, then was separated and purified by reverse phase column chromatography and LH-20 gel column chromatography. Eleven compounds were obtained and identified as Shanzhiganin-1-O-β-D- glucopyranoside methyl ester (1), sucrose (2), oxalic acid (3), 6,7-dihydroxycoumarin (4), caffeic acid (5), luteolin (6), apigenin-7-O-β-D-(-6″-p-coumaroyl)-glucoside (7), protocatechuic acid(8), 6-methyl-iso-phthalic acid Me ester (9), 3,5-dimethoxyl-4-hydroxyl-benzoic acid (10), lamiophlomisine (11). Uniformly, six compounds were obtained by column chromatography and identified on the basis of spectroscopic analysis from n-BuOH fraction: forsythoside B(12), verbascoside(13), luteolin-7-O-[β-D-apiofuranosyl(1→6)]-β-D-glucopyranoside(14),apigenin-7-O-β-D-glucopyranoside (15), hyperin (16), luteolin-7-O-β-D-glucopyranoside (17). Among them, compound 11 is a novel monoterpene alkaloid. In the in vivo hemostatic assay, mice were orally given the aqueous extract at doses of 0.5g·kg- 1, 1.0g·kg-1, 2.0g·kg- 1 and each of the polarity fractions at dose of 0.1g·kg-1. Capillary coagulation time was used to evaluate the hemostatic effects. Results showed that the three doses of aqueous extract could decrease the coagulation time by 23.4%, 26.7% and 41.9%,respectively,and the EtOAC fraction by 18.77%. Aqueous extract of the aerial parts of L. Rotata displayed a dose-dependent hemostatic effect and the active components seem to exist mainly in the EtOAC fraction. Then, hemostatic activity of caffeic acid was estimated at the dose of 117mg/kg in mice and it was observed that caffeic acid could decrease the coagulation time by 26.4%, and seems to be one of the effective hemostatic component of this plant.