Anti-inflammatory activity of Mitraphylline isolated from Uncaria tomentosa bark.

ETHNOPHARMACOLOGICAL RELEVANCE: Uncaria tomentosa (Willd. ex Roem. & Schult.) DC. (Rubiaceae) is widely used by populations living in South America to treat many ailments associated with inflammatory disorders. Mitraphylline was shown to be the major pentacyclic oxindolic alkaloid present in the bark chloroformic extract of this plant. Its activity against cytokines involved in inflammation process was tested in a murine model in vivo. MATERIALS AND METHODS: Mice received mitraphylline once a day for 3 days at 30 mg/kg/day by oral route. Then, they were subjected to bacterial lipopolysaccharide (LPS) endotoxin (15 mg/kg) and the LPS-induced production of 16 different cytokines was determined by Elisa multiplex. Control group received dexamethasone orally at 2mg/kg/day. Toxicity on K565 cells and murine peritoneal macrophages, in vitro, at doses up to 100 µM was monitored by XTT-colorimetric assay. RESULTS AND CONCLUSIONS: For the first time mitraphylline was tested in vivo against a large range of cytokines that play a crucial role in inflammation. Mitraphylline inhibited around 50% of the release of interleukins 1α, 1ß, 17, and TNF-α. This activity was similar to dexamethasone. It also reduced almost 40% of the production of interleukin 4 (IL-4) while the corticoid did not. Lastly it did not show any toxicity on K565 cells nor murine macrophages at doses up to 100 µM.

Antileishmanial sesquiterpene lactones from Pseudelephantopus spicatus, a traditional remedy from the Chayahuita Amerindians (Peru). Part III.

ETHNOPHARMACOLOGICAL RELEVANCE: The study of traditional remedies used by the Chayahuita, an ethnic group from the Peruvian Amazonia, has prompted us to investigate in detail the ethanolic extract of Pseudelephantopus spicatus (Juss. ex Aubl.) C.F. Baker, which has demonstrated strong biological activity towards Leishmania amazonensis. Our goal was to discover the active compound of this plant-based remedy. MATERIALS AND METHODS: A bioguided fractionation of the crude extract was undertaken based on the biological activity recorded against Leishmania amazonensis axenic amastigotes in in vitro bioassays. RESULTS: Three strongly to moderately active compounds were isolated: two hirsutinolides (the 8,13-diacetyl-piptocarphol and the 8-acetyl-13-O-ethyl-piptocarphol) and ursolic acid. IC(50) against Leishmania amazonensis axenic amastigotes are respectively 0.2, 0.37 and 0.99 µM (while IC(50) of amphotericin B is 0.41 µM). These compounds have never been isolated from this plant species, and germacranolides have never been identified as potential antileishmanial agents. CONCLUSIONS: The compounds isolated from Pseudelephantopus spicatus account for the antileishmanial activity of the plant, thus giving support to its use by the Chayahuita in Peru.

Antimalarial activity of simalikalactone E, a new quassinoid from Quassia amara L. (Simaroubaceae).

We report the isolation and identification of a new quassinoid named simalikalactone E (SkE), extracted from a widely used Amazonian antimalarial remedy made out of Quassia amara L. (Simaroubaceae) leaves. This new molecule inhibited the growth of Plasmodium falciparum cultured in vitro by 50%, in the concentration range from 24 to 68 nM, independently of the strain sensitivity to chloroquine. We also showed that this compound was able to decrease gametocytemia with a 50% inhibitory concentration sevenfold lower than that of primaquine. SkE was found to be less toxic than simalikalactone D (SkD), another antimalarial quassinoid from Q. amara, and its cytotoxicity on mammalian cells was dependent on the cell line, displaying a good selectivity index when tested on nontumorogenic cells. In vivo, SkE inhibited murine malaria growth of Plasmodium vinckei petteri by 50% at 1 and 0.5 mg/kg of body weight/day, by the oral or intraperitoneal routes, respectively. The contribution of quassinoids as a source of antimalarial molecules needs therefore to be reconsidered.

Ta'ta', Huayani: perception of leishmaniasis and evaluation of medicinal plants used by the Chayahuita in Peru. Part II.

AIM OF THE STUDY: A knowledge attitude and practice study centred on leishmaniasis and its treatment was performed among the Chayahuita, an Amazonian Peruvian ethnic group living in an endemic area. This study documents traditional Chayahuita plant's use and disease concepts. Also, activity of some medicinal plants used by the Chayahuita is highlighted and discussed. MATERIALS AND METHODS: Ninety-three Chayahuita people were interviewed, following a semi-structured questionnaire focussed on disease knowledge and perception, personal attitude and healing practices. Simultaneously, a collection of plants was performed in different ecotopes, in order to make an extensive inventory of the pharmacopoeia. RESULTS: For the Chayahuita, cutaneous (CL) and muco-cutaneous leishmaniasis (MCL) are considered as diseases of their own, with specific names, aetiologies, mode of transmission. Regarding CL, Chayahuita people consider that the humid characteristic of the skin ulcer is a discriminative fact orienting the diagnostic forTa'ta' (leishmaniasis). Forty-six different species were designated useful against LC and /or MCL (29 species by means of the questionnaire and 27 species when collecting in different ecotopes). Thirty-seven extracts corresponding to 31 species used medicinally were screened in vitro against Leishmania amazonensis axenic amastigotes, assessing their viability by the reduction of tetrazolium salt (MTT). Six species displayed a good activity (10 microg/ml

Ethnopharmacology and malaria: new hypothetical leads or old efficient antimalarials?

New treatments are urgently needed to curb and eradicate malaria in developing countries. As most people living in malarial endemic areas use traditional medicine to fight this disease, why have new treatments not emerged recently from ethnopharmacology-oriented research? The rationale and limitations of the ethnopharmacological approach are discussed in this paper, focusing on ethnopharmacology methodologies and techniques used for assessing botanical samples for their antimalarial properties. Discrepancies often observed between strong ethnopharmacological reputation and laboratory results are discussed, as well as new research perspectives.

Evaluation of the leishmanicidal activity of plants used by Peruvian Chayahuita ethnic group.

AIM OF THE STUDY: A total of 27 ethanolic plant extracts from 27 species were screened for leishmanicidal activity in vitro against Leishmania amazonensis. Most of the selected species (19) are traditionally used by the Chayahuitas, an Amazonian Peruvian ethnic group, to treat skin affections and/or leishmaniasis. MATERIAL AND METHODS: A colorimetric method based on the reduction of tetrazolium salt (MTT) was used to measure the viability of Leishmania amazonensis promastigote and amastigote stages. RESULTS AND CONCLUSIONS: Only the leaves of two species of the Piperaceae family (Piper hispidum Sw., and Piper strigosum Trel.) showed good leishmanicidal activities (IC(50)<10 microg/ml against amastigotes). Roots of Tabernaemontana sananho Ruiz & Pav. (Apocynaceae), together with bark of Vismia tomentosa Ruiz & Pav. (Clusiaceae), fruits of Solanum straminifolium var straminifolium Jacq. (Solanaceae), and stems of Zamia lindenii Regel ex André (Cycadaceae) showed low activity against amastigote stage (IC(50) around 50 microg/ml). Of those only Tabernaemontana sananho displayed also good activity on promastigotes (IC(50)<10 microg/ml). Results are discussed herein, in relation with the traditional use of the plants and compared with other data from the relevant literature.

Quassia amara L. (Simaroubaceae) leaf tea: effect of the growing stage and desiccation status on the antimalarial activity of a traditional preparation.

In French Guiana, Quassia amara L. (Simaroubaceae) leaf tea is a well-known widely used traditional antimalarial remedy. Impact of the vegetal sampling condition on in vivo and in vitro antimalarial activity was assessed. Traditional infusions were prepared with juvenile or mature leaves, both either fresh or dried. Results showed that growing stage and freshness of vegetal material exert a striking effect on antimalarial activity, both in vitro and in vivo. By far, leaf tea made from fresh juvenile (FJ) Quassia amara leaves was the most active. In vitro, active component (simalikalactone D) concentration correlates biological activities, although unexplained subtle variations were observed. In vivo, tea made with dried juvenile (DJ) leaves displays a peculiar behavior, meaning that some components may help simalikalactone D delivery or may be active in vivo only, therefore enhancing the expected curative effect of the traditional preparation.

From Tonic-cups to Bitter-cups: Kwasi bita beker from Suriname Determination, past and present use of an ancient galenic artefact.

In the main markets of Paramaribo (Suriname), many stands offer what is locally called "Bitter-cups", or "Kwasi bita beker", small footed-cups, roughly carved from a whitish wood. The use of these cups is strictly medicinal and it seems to be restricted to Suriname, as they are not found in neighbouring countries (Guyana, French Guiana). The aim of this study was to identify the botanical origin of Bitter-cups still in use in the Saramaka traditional medicine (as information from field people was controversial), and document the ethnopharmacology of this original galenical artefact. Microscopic and high performance liquid chromatography (HPLC) analyses were carried out on Bitter-cup, and anatomical criteria (marginal parenchyma band, size of intervessel and vessel-ray pits, rays width and rays composition, vessels clustering, frequency and size of parenchyma pits) together with HPLC profiles of the macerates showed that the wood cup was similar to Quassia amara L. (Simaroubaceae) wood. Ethnopharmacological investigation indicates that the use of these cups is simply due to the pharmacological properties attributed to "bitters", and is strongly linked to tradition and cultural attitudes. This study also emphasizes the long lasting use of these cups, now restricted to Suriname only, with almost no variation over one century.

Antimalarial activity of some Colombian medicinal plants.

Antimalarial activity of 10 vegetal extracts (9 ethanolic extracts and 1 crude alkaloid extract), obtained from eight species traditionally used in Colombia to treat malaria symptoms, was evaluated in culture using Plasmodium falciparum chloroquine resistant (FcB2) strain and in vivo on rodent malaria Plasmodium berghei. The activity on ferriprotoporphyrin biomineralization inhibition test (FBIT) was also assessed. Against Plasmodium falciparum, eight extracts displayed good activity Abuta grandifolia (Mart.) Sandwith (Menispermaceae) leaves, Acacia farnesiana (L.) Willd. (Mimosaceae) leaves, Acnistus arborescens (L.) Schltdl. (Solanaceae) aerial part, Croton leptostachyus Kunth (Euphorbiaceae) aerial part, Piper cumanense Kunth (Piperaceae) fruits and leaves, Piper holtonii C. DC. (Piperaceae) aerial part and Xylopia aromatica (Lam.) Mart. (Annonaceae) bark with IC(50) values ranging from <1 to 2.1 microg/ml, while in the in vivo model only Abuta grandifolia alkaloid crude extract exhibits activity, inhibiting 66% of the parasite growth at 250 mg/kg/day. In the FBIT model, five extracts were active (Abuta grandifolia, Croton leptostachyus, Piper cumanense fruit and leaves and Xylopia aromatica).

Simalikalactone D is responsible for the antimalarial properties of an Amazonian traditional remedy made with Quassia amara L. (Simaroubaceae).

French Guiana (North-East Amazonia) records high malaria incidence rates. The traditional antimalarial remedy most widespread there is a simple tea made out from Quassia amara L. leaves (Simaroubaceae). This herbal tea displays an excellent antimalarial activity both in vitro and in vivo. A known quassinoid, simalikalactone D (SkD), was identified as the active compound, with an IC(50) value of 10nM against FcB1 Plasmodium falciparum chloroquine resistant strain in vitro. Lastly, it inhibits 50% of Plasmodium yoelii yoelii rodent malaria parasite at 3.7 mg/kg/day in vivo by oral route. These findings confirm the traditional use of this herbal tea.

Validation of use of a traditional antimalarial remedy from French Guiana, Zanthoxylum rhoifolium Lam.

Zanthoxylum rhoifolium bark (Rutaceae) is a medicinal plant, traditionally used in French Guiana to treat and prevent malaria. Bioassay-guided extractions of Zanthoxylum rhoifolium bark have shown that antiplasmodial activity is concentrated in the alkaloid fraction. Further fractionation of this extract has yielded seven benzophenanthridine alkaloids, dihydroavicine 1, dihydronitidine 2, oxyavicine 3, oxynitidine 4, fagaridine 5, avicine 6 and nitidine 7. Antimalarial activity of the last five compounds has been evaluated, and nitidine was the most potent, displaying an IC(50)<0.27microM against Plasmodium falciparum. Investigation of the traditional remedy, a trunk bark decoction in water, has shown that fagaridine 5, avicine 6 and nitidine 7 are also present in the decoction, therefore justifying the traditional use of Zanthoxylumrhoifolium bark as antimalarial.

Antimalarial remedies in French Guiana: a knowledge attitudes and practices study.

A "knowledge attitudes and practices" study about malaria treatments was undertaken in French Guiana, along with an ethnopharmacological study. One hundred and seventeen people from five different groups and nationalities (Creole, Palikur, Galibi, Brazilian, and European) answered the questionnaire. The results were analysed using univariate and multivariate statistical analysis. First, we evaluated the overall knowledge about malaria from the interviewed people. According to bio-medical concepts, we noticed that they have a good knowledge of this illness. Secondly, we studied the treatment used by sick people during their last malaria attack. We demonstrated that, although bio-medical treatment is available in this area, people use both modern drugs and traditional remedies. Finally, preventive attitudes have been examined. One-third of the interviewed people drink regularly some herbal remedy to prevent febrile illnesses and malaria, thus displaying a strong concern about this disease. The ethnopharmacological study highlighted the frequent use of traditional remedies, along with their mode of preparation and administration. A total of 34 different species (both from flora and fauna) have been registered as antimalarial. Twenty-seven are used for curative purposes, 20 as preventive and 13 of them are used for both purposes. Quassia amara (Simaroubaceae) whose antimalarial activity has already been demonstrated was the species most frequently used as antimalarial for curative and preventive purposes.

Evaluation of French Guiana traditional antimalarial remedies.

In order to evaluate the antimalarial potential of traditional remedies used in French Guiana, 35 remedies were prepared in their traditional form and screened for blood schizonticidal activity in vitro on Plasmodium falciparum chloroquine re4sistant strain (W2). Some of these extracts were screened in vivo against Plasmodium yoelii rodent malaria. Ferriprotoporphyrin inhibition test was also performed. Four remedies, widely used among the population as preventives, were able to inhibit more than 50% of the parasite growth in vivo at around 100 mg/kg: Irlbachia alata (Gentiananceae), Picrolemma pseudocoffea (Simaroubaceae), Quassia amara (Simaroubaceae), Tinospora crispa (Menispermaceae) and Zanthoxylum rhoifolium (Rutaceae). Five remedies displayed an IC50 in vitro < 10 microg/ml: Picrolemma pseudocoffea, Pseudoxandra cuspidata (Annonaceae) and Quassia amara leaves and stem, together with a multi-ingredient recipe. Two remedies were more active than a Cinchona preparation on the ferriprotoporphyrin inhibition test: Picrolemma pseudocoffea and Quassia amara. We also showed that a traditional preventive remedy, made from Geissospermum argenteum bark macerated in rum, was able to impair the intrahepatic cycle of the parasite. For the first time, traditional remedies from French Guiana have been directly tested on malarial pharmacological assays and some have been shown to be active.

In vitro immunomodulatory activity of plants used by the Tacana ethnic group in Bolivia.

One hundred and seventy-eight ethanolic plant extracts from the pharmacopoeia of the Tacana, an ethnic group from Bolivia, were screened for immunomodulatory activity using complement cascade inhibition and ADP-induced platelet aggregation inhibition assays. Six impaired both complement pathways (classical and alternative): stem bark from Astronium urundeuvea (Anacardiaceae), Cochlospermum vitifolium (Cochlospermaceae), Terminalia amazonica (Combretaceae), Triplaris americana (Polygonaceae), Uncaria tomentosa (Rubiaceae) and Euterpe precatoria (Arecaceae) roots. Inhibition of complement cascade was independent of essential ion complexation, and was not due to direct hemolytic activity on target red blood cells. For A. urundeuvea, C. vitifolium, and T. amazonica, anti-inflammatory activity relied on cyclo-oxygenase inhibition. Four of these species (A. urundeuva, T. americana, U. tomentosa and E. precatoria) are used traditionally to treat inflammatory processes.

A search for natural bioactive compounds in Bolivia through a multidisciplinary approach. Part VI. Evaluation of the antimalarial activity of plants used by Isoceño-Guaraní Indians.

Seventy-seven plant extracts (corresponding to 62 different species) traditionally used by the Isoceño-Guaraní, a native community living in the Bolivian Chaco, were screened for antimalarial activity in vitro on Plasmodium falciparum chloroquine sensitive strain (F32), and on ferriprotoporphyrin (FP) IX biocrystallisation inhibition test (FBIT). Among these extracts, seven displayed strong in vitro antimalarial activity, and 25 were active in the FBIT test. Positive results on both tests were recorded for six extracts: Argemone subfusiformis aerial part, Aspidosperma quebracho-blanco bark, Castela coccinea leaves and bark, Solanum argentinum leaves and Vallesia glabra bark. Results are discussed in relation with Isoceño-Guaraní traditional medicine. Further studies to be undertaken in relation with these results are also highlighted.

Pharmacopoeia in a shamanistic society: the Izoceño-Guaraní (Bolivian Chaco).

We present the results of an ethnopharmacological research within a Bolivian lowland ethnic group from the dry Chaco, the izoceño-guaraní. Izoceño-guaraní people belong to the extended Chiriguano group. They are actually organised in independent communities, settled down in south-east Bolivia. Struggling very soon for their rights, landowners of their territory, izoceño-guaraní appear to be well organised and maintain a still vivid culture. Medicine is in the hands of Paye who are recognised as specialists in their own group. Ethnopharmacological research leads us to collect approximately over 306 species, 189 of them having medicinal uses. We present here an overview of the izoceño-guaraní ethnomedicine and pharmacopoeia, based on vegetal and animal products.

Actividad citotoxica de plantas medicinales usadas por el grupo étnico boliviano Tacana / Cytotoxic activity of medicinal plants used by the Takana ethnic group in Bolivia

Pregunta de investigación ¿ Las plantas utilizadas en la Medicina Tradicional por los Tacanas manifiestan actividad citotóxica in vitro con el test de la sulforodamina? Objetivos. Determinar la actividad citotóxica in vitro de extractos de plantas utilizadas en la Medicina Tradicional por los Tacanas. Lugar. Población Tacana, IBBA. Métodos. Cultivo de lineas celulares humanas, KB (carcinoma nasofaringeo), HeLa (carcinoma cervical)Ensayo colorimétrico de sulforodamina B, in vitro. Resultados. 118 extractos etanólicos de especies vegetales estudiadas in vitro utilizadando dos lineas tumorales humanas. 14 especies vegetales mostraron actividad citotóxica in vitro. De la 14 especies citotóxicas, 12 especies vegetales presentaron Cl50<- a 10ug/ml. Conclusión. 13 porciento de las plantas estudiadas presentaron actividad citotóxica, pese a no ser seleccionadas por su uso tradicional como anticancerígenos, pues, la actividad anticancer de las plantas estudiadas no consta en la medicina tradicional de los Tacanas. La actividad antitumoral, antioxidante y antimutagénica dae las especies citotóxicas en este trabajo y otras especies diferentes, per, de los mismos géneros estudiados fue reportada por científicos de otras latitudes, lo que nos podría indicar que los principios activos serian comunes en estos géneros, permitiéndonos indentificar nuevas fuentes de medicamentos. Todas las especies activas merecen estudios complementarios.

Actividad antipaludica invitro e in vivo de plantas usadas por el grupo etnico boliviano Tacana / Antimalarial activity in vitro and in vivo of plants used by the ethnical group Tacana Bolivia

Pregunta de investigaciòn. ¿Las palntas utilizadas en la Medicina Tradicional porl os Tacanas especialmente como febrifugas y antimalariales, manifiestan acatividad antipàludica en las pruebas biòlogicas in viatro e in vivo recomendadas por la comunidad cientifica?. Objetivos: Determinar la actividad antipàludica in vitro e in vivo de extractos de plantas utilizadas en Medicina Tradicional por los Tacanas. Lugar: IINSAD, IBBA, Poblaciòn Tacana. Mètodos: Cultivo de estadios eritrocitarios de Plasmodium falciparum. Micromètodo radioisotòpico, in vitro. Test supresivo de 4 dìas in vivo. Resultados: 125 especies vegetales estudiadas in vitro e in vivo utilizando cepas cloroquina-sensibles y cloroquina-resistentes de plasmodium falciparum y ratones infectados con Plasmodium berghei NK65. De las 10 especies que se usan como plantas febrifugas, una mostrò actividad in vivo. 7 extractos etanòlicos de las especies vegetales: Bowdichia virgilioides, Caesalpinia pluviosa, Licaria canella, Nectandra aff. hihua, Protium glabrescens, Gymnosporia sp. y Quina Florida, fueron los màs activos in vitro e in vivo. Conlusión: De las 7 esoecies vegetales que mostraron actividad in vitro o in vivo, sòlo una especie Bawdichia virgilioides fue indicada especificamente para el tratameinto de malaria, su uso tradicional está ampliamente respaldado por las diferentes pruebas antipalúdicas realziadas en este trabajo, B. virgilioides es una fuente potencial de drogas antimalariales, es una prioridad futuras investigaciones clínicas a través de diferentes proyectos. Caesalpinia pluviosa, licaria canella, Nectandra aff, hihua, Protium glabrescens, Gymnosporia sp y Quina florida merecen estudios complementarios.

A search for natural bioactive compounds in Bolivia through a multidisciplinary approach. Part V. Evaluation of the antimalarial activity of plants used by the Tacana Indians.

One hundred and twenty-five extracts of 122 different plant species traditionally used by the Tacana, a native community living in lowland forest at the base of the last foothills of the Cordillera Oriental of the Bolivian Andes, were screened for antimalarial activity in vitro on Plasmodium falciparum chloroquine resistant (D2) and sensitive strains (F32), and were evaluated in vivo on rodent malaria Plasmodium berghei. Five ethanolic stembark extracts showed marked activity either in vitro or in vivo, and only one of them, Bowdichia virgilioides being traditionally used against malaria, was active in vitro (IC50=1 microg/ml on both strains) and in vivo (51% at 100 mg/kg). Other active extracts were from Caesalpinia pluviosa bark displaying activity in vitro against chloroquine resistant strain (IC50 8.3 microg/ml), traditionally used against dysentery; two Lauraceae bark extracts, Nectandra aff. hihua and Licaria canella respectively used for construction purposes and against stomach ache, both displaying activity in vitro against P. falciparum sensible and resistant strains (IC50 around 4 microg/ml); finally, the bark of a strongly aromatic Burseraceae, Protium glabrescens exuding an anti-inflammatory and analgesic resin, was active in vivo only (61% at 100 mg/kg). Results are discussed in relation with Tacana traditional medicine.

A search for natural bioactive compounds in Bolivia through a multidisciplinary approach. Part IV. Is a new haem polymerisation inhibition test pertinent for the detection of antimalarial natural products?

The search for new antimalarial agents in plant crude extracts using traditional screening tests is time-consuming and expensive. New in vitro alternative techniques, based on specific metabolic or enzymatic process, have recently been developed to circumvent testing of antimalarial activity in parasite culture. The haem polymerisation inhibition test (HPIA) was proposed as a possible routine in vitro assay for the detection of antimalarial activity in natural products. A total of 178 plant extracts from the Pharmacopeia of the Bolivian ethnia Tacana, were screened for their ability to inhibit the polymerisation of haematin. Five extracts from Aloysia virgata (Ruíz & Pavón) A.L. Jussieu (Verbenaceae), Bixa orellana L. (Bixaceae), Caesalpinia pluviosa D.C. (Caesalpiniaceae), Mascagnia stannea (Griseb) Nied. (Malpighiaceae) and Trichilia pleenea (Adr. Jussieu) (Meliaceae) demonstrated more than 70% inhibition of haematin polymerisation at 2.5 mg/ml. The extracts were also tested for antimalarial activity in culture against F32 strain (chloroquine-sensitive) and D2 strain (chloroquine-resistant) of Plasmodium falciparum and in vivo against P. berghei. The extract from Caesalpinia pluviosa was the only one that showed activity in HPIA and in the classical test in culture. The accuracy and pertinence of HPIA, applied to natural products is discussed.