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1.
J Helminthol ; 85(3): 304-12, 2011 Sep.
Article in English | MEDLINE | ID: mdl-20863422

ABSTRACT

Ethanolic and aqueous extracts of selected medicinal plants from Cameroon and Ghana were assessed for their in vitro anthelmintic activity by using the bovine filarial parasite Onchocerca ochengi and the free living nematode Caenorhabditis elegans, a model organism for research on nematode parasites. Worms were incubated in the presence of different concentrations of extracts and inhibitory effects were monitored at different time points. Among the extracts used in this study, ethanolic extracts of Anogeissus leiocarpus, Khaya senegalensis, Euphorbia hirta and aqueous extracts from Annona senegalensis and Parquetina nigrescens affected the growth and survival of C. elegans and O. ochengi significantly. The mortality was concentration dependent with an LC50 ranging between 0.38 and 4.00 mg/ml for C. elegans (after 72 h) and between 0.08 and 0.55 mg/ml for O. ochengi after a 24 h incubation time. Preliminary phytochemical screenings on these extracts revealed the presence of flavonoids, alkaloids, saponins, carbohydrates and tannins in the extracts. Accordingly, application of A. leiocarpus, K. senegalensis, E. hirta and A. senegalensis extracts could provide alternatives in the control of helminthic infections.


Subject(s)
Anthelmintics/pharmacology , Caenorhabditis elegans/drug effects , Onchocerca/drug effects , Plant Extracts/pharmacology , Plants, Medicinal/chemistry , Animals , Anthelmintics/chemistry , Anthelmintics/isolation & purification , Caenorhabditis elegans/growth & development , Cameroon , Ghana , Onchocerca/growth & development , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Survival Analysis
2.
J Ethnopharmacol ; 174: 628-36, 2015 Nov 04.
Article in English | MEDLINE | ID: mdl-26087235

ABSTRACT

ETHNOPHARMACOLOGICAL RELEVANCE: Leaves from Combretum mucronatum Schum. & Thonn. are traditionally used for wound healing in Western Africa. Aqueous and hydroalcoholic extracts of the dried leaves recently have been shown to stimulate viability of human keratinocytes and dermal fibroblasts. AIM OF THE STUDY: Phytochemical characterization of the herbal material, development of a validated HPLC methodology for quality control, and pinpointing the underlying pharmacological mechanism under in vitro conditions to understand the impact of C. mucronatum extracts on human skin cells. MATERIALS AND METHODS: Extracts obtained from the leaves from C. mucronatum by using solvents with different polarities (petrol ether, dichloromethane, ethanol-water 50%, water) were investigated concerning phytochemical composition by GC-MS, LC-MS and in part after fractionation and isolation of purified compounds. For quality control of the herbal material an ICH-2 validated UHPLC method was developed for quantification of the lead compounds epicatechin, procyanidin B2, vitexin and isovitexin. In vitro studies were performed using HaCaT keratinocyte cell line, primary keratinocytes and primary skin fibroblasts with determination of viability (MTT assay), cell proliferation (BrdU incorporation ELISA), cell toxicity (LDH release) and keratinocyte differentiation, using involucrin and keratin K10 as differentiation marker (confocal laser scanning microscopy, Western blot). RESULTS: A detailed phytochemical composition analysis of the extracts from the leaves from C. mucronatum was performed (compounds 1-34) and epicatechin, procyanidin B2, vitexin and isovitexin are assessed to be the lead compounds of the polar extract. Quantitative UHPLC investigations indicated mature leaves to have higher polyphenol content in comparison to young leaves. The drying process of the plant material was shown to have great influence on the content of the lead compounds. The aqueous extract (0.1-100µg/mL) did not change cell viability of dermal fibroblasts and keratinocytes but inhibited cellular proliferation rates significantly at 100µg/mL. The extract stimulated cellular differentiation of primary keratinocytes significantly at 1 and 10µg/mL. Procyanidin B2 at 1 and 10µM was shown to be responsible for the induction of this cellular differentiation, while epicatechin, and procyanidins B5, C1 and D1 were inactive. CONCLUSION: The in vitro effects of the aqueous extract on the skin cells rationalized the remedial effect in wound healing and possibly accounts for the reason why this plant may be widely used for this purpose. On the basis of this study extracts from the leaves of C. mucronatum therefore have potential for the use in wound healing.


Subject(s)
Cell Differentiation/drug effects , Combretum/chemistry , Plant Extracts/pharmacology , Plant Leaves/chemistry , Proanthocyanidins/pharmacology , Wound Healing/drug effects , Cell Line , Cell Proliferation/drug effects , Fibroblasts/drug effects , Ghana , Humans , Keratinocytes/drug effects , Proanthocyanidins/isolation & purification , Skin/cytology , Tannins/analysis
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