lancemaside-a and echinocystic-acid

The rhizome of Codonopsis lanceolata (family Campanulaceae), which contains lancemaside A as a main constituent, is frequently used in the traditional Chinese medicine for the treatment of inflammatory diseases. Lancemaside A exhibits anti-inflammatory effect in vitro and in vivo. However, orally administered lancemaside A is metabolized to echinocystic acid by the intestinal microflora and the metabolite is absorbed into the blood. Therefore, to understand whether echinocystic acid is effective against skin inflammatory diseases, we assessed its inhibitory effect against 12-O-tetra decanoylphorbol-13-acetate (TPA)-induced ear inflammation in mice. Topically administered echinocystic acid potently suppressed TPA-induced ear swelling. The suppression rates at 0.05 and 0.10 % concentrations were 65 and 73 %, respectively. Echinocystic acid also inhibited TPA-induced myeloperoxidase activity, as well as COX-2, iNOS, TNF-α and IL-1β expressions. Echinocystic acid inhibited NF-κB in TPA-treated mouse ears, as well as in lipopolysaccharide-stimulated peritoneal macrophages. Its potency is comparable with that of dexamethasone. These findings indicate that echinocystic acid may ameliorate inflammatory diseases, such as dermatitis.

Topics: Animals; Anti-Inflammatory Agents; Cells, Cultured; Dermatitis; Macrophages, Peritoneal; Male; Mice; Mice, Inbred ICR; Microscopy, Confocal; Microscopy, Fluorescence; Molecular Structure; Oleanolic Acid; Saponins; Spectrometry, Mass, Electrospray Ionization; Tetradecanoylphorbol Acetate

The rhizome of Codonopsis lanceolata (CL, family Campanulaceae), of which the main constituent is lancemaside A, has been used for cough and bronchitis in traditional Chinese medicine. To evaluate anti-colitic effect of CL, we examined anti-inflammatory effect of CL extracts, lancemaside A and its metabolites in lipopolysaccharide (LPS)-stimulated peritoneal macrophages and 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitic mice. Among CL extracts, CL BuOH extract inhibited LPS-induced IL-1β, IL-6 and TNF-α expression, as well as NF-κB activation most potently. CL BuOH extract also inhibited colon shortening and myeloperoxidase activity in TNBS-induced colitic mice. Among lancemaside A, a main constituent of CL BuOH extract, and its metabolites (lancemaside X, echinocystic acid-3-O-β-d-glucopyranoside and echinocystic acid), echinocystic acid inhibited the expression of the pro-inflammatory cytokines, IL-1β, IL-6, and TNF-α, as well as the phosphorylation of IKKβ and p65 in LPS-stimulated peritoneal macrophages most potently. Echinocystic acid also potently inhibited the binding of LPS to TLR4 on peritoneal macrophages. Lancemaside A and its metabolite, echinocystic acid, inhibited TNBS-induced colonic inflammation, including colon shortening, increased myeloperoxidase activity and pro-inflammatory cytokine expression, and NF-κB activation in mice. The anti-colitic effect of echinocystic acid was superior to that of lancemaside A. Based on these findings, orally administered lancemaside A may be metabolized to echinocystic acid, which may express anti-colitic effect by inhibiting the binding of LPS to TLR4 on the macrophages.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Cells, Cultured; Codonopsis; Colitis; Colon; Cytokines; Humans; Inflammation Mediators; Lipopolysaccharides; Macrophages, Peritoneal; Male; Medicine, Chinese Traditional; Mice; Mice, Inbred ICR; NF-kappa B; Oleanolic Acid; Protein Binding; Rhizome; Saponins; Signal Transduction; Toll-Like Receptor 4; Trinitrobenzenesulfonic Acid

Lancemaside A, a triterpenoid saponin isolated from the roots of Codonopsis lanceolata, has been reported to ameliorate the reduction of blood testosterone levels induced by immobilization stress in mice. In the present study, we investigated the metabolism and absorption of lancemaside A in mice. After oral administration of lancemaside A at 100 mg/kg body weight, the unmetabolized compound appeared rapidly in plasma (t (max) = 0.5 h). Lancemaside A has a low bioavailability (1.1%) because of its metabolism by intestinal bacteria and its poor absorption in the gastrointestinal tract. Furthermore, we identified four metabolites from the cecum of mice after oral administration of lancemaside A: codonolaside II, echinocystic acid, echinocystic acid 28-O-beta-D: -xylopyranosyl-(1 --> 4)-alpha-L: -rhamnopyranosyl-(1 --> 2)-alpha-L: -arabinopyranosyl ester, and echinocystic acid 28-O-alpha-L: -rhamnopyranosyl-(1 --> 2)-alpha-L: -arabinopyranosyl ester. Among these metabolites, codonolaside II and echinocystic acid were detected in plasma, and their t (max) values were 4 and 8 h, respectively. These findings should be helpful for understanding the mechanism of the biological effect of lancemaside A.

Topics: Administration, Oral; Animals; Cecum; Chromatography, Liquid; Codonopsis; Esters; Male; Mass Spectrometry; Mice; Molecular Structure; Oleanolic Acid; Plant Roots; Saponins

A high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) method employing electrospray ionization (ESI) has been developed for simultaneous determination of lancemaside A (3-O-beta-D-glucuronopyranosyl-3beta, 16alpha-dihydroxyolean-12-en-28-oic acid 28-O-beta-D-xylopyranosyl(1-->3)-beta-D-xylopyranosyl-(1-->4)-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranosyl ester) and its metabolites in mouse plasma. When lancemaside A (60 mg/kg) was orally administered to mice, echinocystic acid was detected in the blood. T(max) and C(max) of the echinocystic acid were 6.5+/-1.9 h and 56.7+/-29.1 ppb. Orally administered lancemaside A was metabolized to lancemaside X (3beta, 16alpha-dihydroxyolean-12-en-28-oic acid 28-O-beta-D-xylopyranosyl(1-->3)-beta-D-xylopyranosyl-(1-->4)-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranosyl ester) by intestinal microflora in mice, which was metabolized to echinocystic acid by intestinal microflora and/or intestinal tissues. Human intestinal microflora also metabolized lancemaside A to echinocystic acid via lancemaside X. These results suggest that the metabolism by intestinal microflora may play an important role in pharmacological effects of orally administered lancemaside A.

Topics: Animals; Bacteria; Chromatography, Liquid; Feces; Gastrointestinal Tract; Humans; Male; Metagenome; Mice; Mice, Inbred ICR; Oleanolic Acid; Reproducibility of Results; Saponins; Spectrometry, Mass, Electrospray Ionization; Tandem Mass Spectrometry