SEDUM TELEPHIUM L.: FROM TRADITIONAL USE TO PHARMACOLOGICAL RESEARCH INTRODUCTION CHEMICAL CHARACTERIZATION ANTIOXIDANT ACTIVITY CONCLUSIONS Sedum telephium L. has been used in clinical practice for many years to treat skin wounds. Despite the consolidated effectiveness, its exact mechanism of action in wound healing was largely unknown. In our recent studies, S. telephium juice proved to be effective on all the phases of tissue regeneration. At 0,1% w/w, sedum juice was found to be a strong antiradicalic agent and modulated the immunitary response upregulating important chemotactic chemokines. At the same concentration, sedum juice increased collagen neosynthesis and upregulated the expression of sirtuin 1 in human fibroblasts. Finally, sedum juice at 0,1% w/w resulted significantly effective in the wound healing assay, compared to control. Those pharmacological features are attributed to its phytocomplex, rich in flavonoids and polysaccharides. REFERENCES [1] M. Biagi, E. Miraldi, B. Barlozzini (2013) Piante Medicinali, 12: 6-9. [2] PM. Guarrera (2006) Usi e Tradizioni della Flora Italiana. Medicina popolare ed etnobotanica. Aracne, Roma. [3] S. Vitalini, C. Puricelli, I. Mikerezi, et al. (2015) J. Ethnopharmacol., 173: 435– 458. [4] S. Balatri (2013) Piante Medicinali, 12: 3-5. [5] F. Bonina, C. Puglia, A. Tomaino et al. (2000) J. Pharm. Pharmacol., 52: 1279- 1285. [6] X.Z. Bai, J.Q. Liu, L.L. Yang et al. (2016) Br J Pharmacol., 173: 1589-1601. MARCO BIAGI 1 , ELISABETTA MIRALDI 1 , DANIELA GIACHETTI 1 , EFTHYMIA E. TSIOUTSIOU 1 , PAOLO GOVERNA 1 , PAOLO GIORDANI 2 , LAURA CORNARA 3 1 DSFTA, University of Siena, Via Laterina 8, 53100 Siena, Italy 2 DIFAR,University of Genova, Viale Cembrano 4, 16148 Genova, Italy 3 DISTAV, University of Genova, Corso Europa 26, 16132 Genova, Italy 111° Congresso della Società Botanica Italiana III International Plant Science Conference (IPSC), Rome 21-23 September 2016 The species has been frequently used for various kinds of indications, including the treatment of insect bites, burns, corns, ulcers, warts, abscesses and wounds [3]. The fresh leaves are applied as poultice for topical application, whereas the juice from crushed fresh leaves is used to heal scars [1]. In modern herbal medicine, S. telephium has been used in clinical practice even in serious emergency cases, such as for treating open wounds and deep burns of II and III degree, or for major damage from traumatic events involving bone [4]. The main chemical constituents of the fresh juice of S. telephium are represented by polyphenols and more than 5% of soluble polysaccharides [5]. In this poster we present the recent acquisitions on phytochemistry, the biological properties and mechanism of action of S. telephium fresh juice using human fibroblasts and keratinocytes as well as peripheral blood mononuclear cells in vitro investigations. Sedum telephium L. is one of those species that have received the attention of scientific research many years after its clinical use that derived from centuries of tradition and descriptions provided by various ancient medical texts [1]. Although it has been mostly cultivated for ornamental use, it was also widely mentioned in the Italian ethnobotany. The traditional use of the species has been reported throughout Italy, from Alpine regions to Central and Southern areas of the peninsula [2]. Chemical constituents Relative % of flavonoid fraction kaempferol-3-O-glucoside-7-O-rhamnoside 18% - 42% kaempferol-3,7-dirhamnoside 24% - 29% kaempferol-3-O-neoesperoside-7-O-rhamnoside 12% - 31% quercetin-3-O-glucoside-7-O-rhamnoside 6% - 9% quercetin-3-O-rhamnoside-7-O-glucoside 4% - 5% quercetin-3-O-neoesperoside-7-O-rhamnoside 2% - 3% quercetin-3-O-rhamnoside 0,5% - 1% kaempferol aglycone < 0,5% quercetin aglycone < 0,5% Table 2: Main flavonols and glycosil derivatives in S. telephium Sample Total polyphenols Total flavonoids expressed as quercetin Sedum juice 0,13% 0,10% Total polyohenols content was evaluated through the spetrophotometric Folin-Ciocalteu method. Total flavonoids were determinated according to European Pharmacopoeia 8 th edition. Qualitative flavonoids determination was carried out by means of HPLC-DAD-MS. Sedum juice contains an interesting amount of polyphenols, the majority of which is represented by flavonoids (Table 1). The main flavonols in S. telephium are represented by kaempferol and quercetin derivatives (Table 2). Table 1: Total polyphenols and flavonoids content in S. telephium juice. IMMUNOMODULATORY ACTIVITY OF S. TELEPHIUM L. JUICE ON PBMC Human peripheral blood mononuclear cells (PBMC) of healty volunteers were treated with sedum for 24 hours. Cytokines release was measured by means of non-competitive Elisa. Sedum juice 0.1% modulates IL-1β, IL-6 and, particularly, IL-8 release. WOUND HEALING ASSAY Sample IC 50 Sedum juice 0,13% DPPH TEST DPPH test was used as a rapid and reproducible test to investigate antiradicalic capacity of S. telephium juice that resulted positively affected by the presence of large quantity of flavonoids. SIRT-1 EXPRESSION ON HUMAN FIBROBLAST By means of Western Blot analyses we investigated how S. telephium juice is effective in modulating cell signaling in human fibroblasts. We first studied sirtuin 1 expression as a pivotal cell repairing signal and a new drug target for skin healing [6]. The protein sirtuin 1 resulted upregulated by S. telephium juice. This finding indicates that sirtuin 1 upregulation is a cytoprotective mechanism of sedum juice. SAMPLE COLLECTION AND PREPARATION Sedum telephium L. plants were collected in Siena (Tuscany), in July, that represents the balsamic period of this species. Taxa were identified in the laboratory according to more recent taxonomic revision. The sedum juice was obtained from fresh leaves, by means of a centrifugal juicer, and analyzed and used immediately after centrifugation. 0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 Ctrl Sedum 0.001% Sedum 0.01% Sedum 0.1% SIRT-1 expression versus control * 0 1 2 3 4 5 IL-1β IL-6 IL-8 TNF-α Ctrl Sedum 0.1% Sedum 0.01% Sedum 0.001% 0 50 100 150 200 250 Ctrl Sedum 0.001% Sedum 0.01% Sedum 0.1% PINP % versus control * COLLAGEN-1 NEOSYNTHESIS Wound healing assay on human keratinocytes treated with sedum juice 0.1% for 24h. T0 24h -45% Collagen-1 synthesis was evaluated by means of Western Blot analysis. Sedum juice 0.1% statistically stimulates the collagen neosynthesis by almost 70%.