Antiprotozoal Activity of Plants Used in the Management of Sleeping Sickness in Angola and Bioactivity-Guided Fractionation of Brasenia schreberi J.F.Gmel and Nymphaea lotus L. Active against T. b. rhodesiense.

Folk medicine is widely used in Angola, even for human African trypanosomiasis (sleeping sickness) in spite of the fact that the reference treatment is available for free. Aiming to validate herbal remedies in use, we selected nine medicinal plants and assessed their antitrypanosomal activity. A total of 122 extracts were prepared using different plant parts and solvents. A total of 15 extracts from seven different plants exhibited in vitro activity (>70% at 20 µg/mL) against Trypanosoma brucei rhodesiense bloodstream forms. The dichloromethane extract of Nymphaea lotus (leaves and leaflets) and the ethanolic extract of Brasenia schreberi (leaves) had IC50 values ≤ 10 µg/mL. These two aquatic plants are of particular interest. They are being co-applied in the form of a decoction of leaves because they are considered by local healers as male and female of the same species, the ethnotaxon "longa dia simbi". Bioassay-guided fractionation led to the identification of eight active molecules: gallic acid (IC50 0.5 µg/mL), methyl gallate (IC50 1.1 µg/mL), 2,3,4,6-tetragalloyl-glucopyranoside, ethyl gallate (IC50 0.5 µg/mL), 1,2,3,4,6-pentagalloyl-ß-glucopyranoside (IC50 20 µg/mL), gossypetin-7-O-ß-glucopyranoside (IC50 5.5 µg/mL), and hypolaetin-7-O-glucoside (IC50 5.7 µg/mL) in B. schreberi, and 5-[(8Z,11Z,14Z)-heptadeca-8,11,14-trienyl] resorcinol (IC50 5.3 µg/mL) not described to date in N. lotus. Five of these active constituents were detected in the traditional preparation. This work provides the first evidence for the ethnomedicinal use of these plants in the management of sleeping sickness in Angola.

Use of herbal remedies in the management of sleeping sickness in four northern provinces of Angola.

ETHNOPHARMACOLOGICAL RELEVANCE: This study reports for the first time on the use of folk medicine to treat sleeping sickness and its symptoms in four endemic provinces in northern Angola. By interviewing both traditional practitioners and confirmed patients, it highlights reasons to recourse to folk medicine, the plant species used for this affection as well as arises awareness about the use of particular plants showing potential risks. AIM OF THE STUDY: The aims of this explorative study were three-fold. Firstly, it informed on access to, and use of plant-based medicine as first-choice treatment by infected persons. Secondly, it aimed at collecting comprehensive data from patients and traditional healers on herbal remedies in order to identify plant species used in the management of the disease. Thirdly, it served as contribution for primary indication of potential risk of use associated with the studied plants and their preparation. MATERIALS AND METHODS: The study was conducted in 4 endemic provinces of Angola, namely Bengo, Zaire, Kwanza Norte and Uíge. We explored the use of herbal remedies by conducting structured and semi-structured interviews within two distinct study populations. The first group comprises 30 patients who had been diagnosed for trypanosomiasis and treated by the reference treatment. The second group included 9 traditional practitioners who had already treated sleeping sickness. The plants that were cited during the interviews were collected during field walks under supervision of a traditional healer, then authenticated and deposited at the National Herbarium in Luanda. RESULTS: Of the 30 included patients, 12 (40%) had turned to folk medicine in the management of trypanosomiasis and related symptoms. 7 medicinal plants were reported by this group. Considering the key motivation to consult a traditional practitioner, two main factors accounted for half of the cases: "past experience with folk medicine" and "family habit". Out of 9 traditional practitioners' interviewees, 26 medicinal plants were cited. Roots and leaves were the most used plant parts, and decoction was the common mode of preparation. Evidence for antitrypanosomal activity in the scientific literature was found for 56% (17 of 30) of the identified plant species. The most cited plant was Crossopteryx febrifuga (UR = 6). Some of the cited plants, as for example Aristolochia gigantea, raised concern about potential toxicity. CONCLUSIONS: With 40% of infected persons having turned first to folk medicine before consulting a medical doctor, this explorative study points out that plant-based medicines play an important role in local dynamics of health care. It highlights the need for primary assessment of potential risk of use related to the herbal recipes, and for reporting it to the concerned population. This first ethnobotanical study on trypanosomiasis in endemic provinces of Angola provides information on 30 plants, of which some had been identified as promising for further pharmacological research. Our results provide a first step towards the validation and valorization of Angolan herbal remedies for sleeping sickness.