Planta Med 2009; 75 - PI3
DOI: 10.1055/s-0029-1234767

Analysis of Rhaponticum carthamoides (Willd.) Iljin crude extracts composition and ability to simulate cell proliferation

E Biskup 1, M Golebiowski 2, K Borsuk 2, P Stepnowski 2, E Lojkowska 1
  • 1Department of Biotechnology, Intercollegiate Faculty of Biotechnology University of Gdansk & Medical University of Gdansk, Kladki 24, 80–822 Gdansk, Poland
  • 2Department of Environmental Analytics, Faculty of Chemistry, University of Gdansk, Sobieskiego 18/19, 80–952 Gdansk, Poland

The presented study concerns the activity of leaves crude extracts, isolated from ecdysteroid-rich plant Rhaponticum carthamoides (Willd.) Iljin. Plant material, collected in Koriażma (Archangielsk district), was obtained through the courtesy of FITOSTAR™ company. Composition analysis of chloroform, methanol and water extracts was performed using GC-MS. The presence of plant sterols: campesterol, β-sitosterol and stigmasterol were previously observed in extract of R. carthamoides seeds [1], whereas α- and β-amyrin were detected in this plant for the first time. The ability of R. carthamoides crude extracts, α-amyrin, β-sitosterol and stigmasterol to stimulate the proliferation of human keratinocytes (HaCaT cell line) was analyzed.

Keratinocytes were grown in the presence of analytes (10–40µg/ml of pure compounds or 10–80µg/ml of crude extracts). The ability to influence cell growth and proliferation was determined using methylene blue assay (after 24 and 48h of incubation) and BrdU incorporation assay (after 24h of incubation). Both sterols and α-amyrin enhanced the proliferation of HaCaT cells up to 30% after 48 hours of incubation (methylene blue assay) and up to 60% after 24h incubation (BrdU incorporation assay). Cholesterol (10–40µg/ml), serving as a control, stimulated cell proliferation in a dose response manner (up to 300% after 24h; BrdU assay). Chloroform crude leaves extract inhibited cell growth and turned to be highly cytotoxic even in the lowest concentrations applied (10µg/ml). Methanol extract exhibited mild cytotoxicity at 80µg/ml, whereas water extract did not influence cell growth.

According to our knowledge the ability of α-amyrin, β-sitosterol and stigmasterol to enhance human cells proliferation was shown for the first time. Obtained data indicate that the other component(s) of R. carthamoides can in certain conditions mask the biological activity of the sterols present in their leaves tissue.

Acknowledgements: this research was supported by the European Union within the European Social Fund in the framework of the project "InnoDoktorant – Scholarships for PhD students, I edition" and by the Polish Ministry of Research and Higher Education. Projects numbers: DS 8200–4-0085–9 and N302 3566 33.

Reference: [1] Stránsky, K. et al. (1998) Russ. J. Plant Physiol. 45:333–338.