furosin and Geraniin

Leaves from Phyllanthus muellerianus (Kuntze) Exell. are traditionally used for wound healing in Western Africa. Aqueous extracts of dried leaves recently have been shown to stimulate proliferation of human keratinocytes and dermal fibroblasts. Within bioassay-guided fractionation the ellagitannins geraniin (1), corilagin (2), furosin (3), the flavonoids quercetin-3-O-β-D-glucoside (isoquercitrin), kaempferol-3-O-β-D-glucoside (astragalin), quercetin-3-O-D-rutinoside (rutin), gallic acid, methyl gallate, caffeic acid, chlorogenic acid, 3,5-dicaffeoylquinic acid and caffeoylmalic acid (phaselic acid) have been identified in P. muellerianus for the first time. Geraniin was shown to be the dominant component of an aqueous extract. Suitable analytical methods for quality control of geraniin in P. muellerianus extract (methanol/water, 70/30) have been developed and validated based on ICH guidelines (ICH-compliant protocol). Geraniin and furosin increased the cellular energy status of human skin cells (dermal fibroblasts NHDF, HaCaT keratinocytes), triggering the cells towards higher proliferation rates, with fibroblasts being more sensitive than keratinocytes. Highest stimulation of NHDF by geraniin was found at 5 μM, and of keratinocytes at 50-100 μM. Furosin stimulated NHDF at about 50 μM, keratinocytes at about 150-200 μM. Necrotic cytotoxicity of geraniin, as measured by LDH release, was observed at 20 μM for NHDF and 150 μM for keratinocytes. Toxicity of furosin--less than that of geraniin--was observed at > 400 μM. Furosin and geraniin stimulated the biosynthesis of collagen from NHDF at 50 μM and 5-10 μM respectively. Geraniin at 105 μM significantly stimulated the differentiation in NHEK while furosin had a minor influence on the expression of involucrin and cytokeratins K1 and K10. The study proves clearly that hydrophilic extracts from P. muellerianus and especially the lead compound geraniin exhibit stimulating activity on dermal fibroblasts and keratinocytes, leading to increased cell proliferation, barrier formation and formation of extracellular matrix proteins. From these findings the traditional clinical use of such extracts for wound healing seems to be justified.

Topics: Cell Differentiation; Chromatography, High Pressure Liquid; Collagen; Fibroblasts; Flavonoids; Glucosides; Humans; Hydrolyzable Tannins; Keratinocytes; Medicine, African Traditional; Phyllanthus; Plant Extracts; Plant Leaves; Plants, Medicinal; Skin; Wound Healing

The present study describes the occurrence of two ellagitannins in the ethanolic extract of the leaves and stems of Phyllanthus sellowianus (Euphorbiaceae). Their preliminary antinociceptive properties were also evaluated. The two ellagitannins were identified on the basis of 1H- and 13C-NMR spectra data and by mixed co-TLC and co-HPLC injection with an authentic sample of furosin and geraniin. Preliminary pharmacological analysis revealed that both furosin and geraniin (3 to 30 mg/kg, i.p.), given 30 min before testing, exhibited significant and dose-related antinociceptive properties against acetic acid-induced abdominal constrictions in mice. Geraniin and furosin were about six- to seven-fold more potent at the ID50 level (micromol/kg) as analgesics than aspirin and acetaminophen, respectively, although they were less efficacious when compared with the standard drugs. These data extend our previous studies and indicate that the two ellagitannins isolated from P.sellowianus, identified as furosin and geraniin are, at least in part, responsible for the antinociceptive actions reported previously for the hydroalcoholic extract of P.sellowianus and other plants belonging to the genus Phyllanthus.

Topics: Abdominal Pain; Acetaminophen; Acetates; Acetic Acid; Analgesics; Animals; Aspirin; Brazil; Glucosides; Hydrolyzable Tannins; Male; Medicine, Traditional; Mice; Molecular Structure; Plants, Medicinal; Tannins