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Potential of a Khaya ivorensis -Alstonia boonei extract combination as antimalarial prophylactic remedy
Authors:Tepongning Roselyne Nzangue  Lucantoni Leonardo  Nasuti Cinzia Carla  Dori Geme Urge  Yerbanga Serge Rakiswende  Lupidi Giulio  Marini Carlotta  Rossi Giacomo  Esposito Fulvio  Habluetzel Annette
Affiliation:a University of Camerino, School of Pharmacy, Piazza dei Costanti, 62032 Camerino (MC), Italy
b University of Camerino, School of Science and Biotechnology, Via Gentile III Da Varano, 62032 Camerino, Italy
c University of Camerino, School of Veterinary Sciences, Via Circonvallazione, 93/95 Matelica, Italy
Abstract:

Ethnopharmacological relevance

The decoction of the combined stem barks of Khaya ivorensis A. Chev. (Meliaceae) and Alstonia boonei De Wild (Apocynaceae) has a history of use in traditional medicine of central Cameroon for malaria treatment but also for the prevention of the disease.

Aim of the study

The purpose of this investigation was to determine the antiplasmodial activity of Khaya ivorensis (K) and Alstonia boonei (A) preparations in the murine malaria model Plasmodium berghei/Anopheles stephensi, to estimate their prophylactic potential and to assess acute and sub-acute toxicity of the formulations prepared according to the traditional recipes.

Materials and methods

Aqueous extracts from the stem-bark of the two plants were prepared and tested separately and in combination. BALB/c mice were treated for 9 days and challenged on day 3 by exposure to mosquitoes infected with Plasmodium berghei. Treatment doses ranged between 200 and 400 mg/kg/day, corresponding approximately to the dosage applied by traditional healers to cure malaria patients or prevent the disease. Parasitemia reduction in treated animals was calculated from Giemsa smear counts, of two replicate experiments. To estimate acute toxicity in terms of median lethal dose (LD50), geometrically increasing doses were administered to mice. Sub-acute toxicity of the herbal combination (KA) was investigated by administering the same doses as in the antiplasmodial activity test for a period of 14 days, followed by 14 days of recovery observation. Locomotor activity (Open Field Test), body weight, liver and kidney morphology were monitored.

Results

The combination KA was found to exhibit antiplasmodial activity in the murine malaria model. In mice treated with the combination remedy at a dosage of 200 mg/kg/day, parasitemia values of 6.2% ± 1.7 and 6.5% ± 0.8 were recorded, compared to 10.8% ± 1.3 and 12.0% ± 4.0 in controls (p < 0.01). Doubling the dosage of the extracts did not significantly increase parasite suppression. When extracts of K and A were administered separately at a dosage of 400 mg/kg, a reduction in parasitemia was still obtained, but it did not reach statistical significance. Toxicity studies yielded comforting results: the LD50 was estimated to be greater than 2779.5 mg/kg. Moreover, mice exposed to the fourteen-day repeated-dose toxicity test (sub-acute toxicity test) did not display weight loss, liver or kidney morphological modifications, significant alterations in locomotor activity or any other sign of illness.

Conclusion

The antiplasmodial activity and the wide dose interval between the therapeutic dosage and the toxic dosage exhibited by the KA herbal combination in the murine malaria model argue in favor of its use as an antimalarial prophylactic remedy. It remains to be demonstrated by human clinical trials whether the combination remedy, when taken by inhabitants during malaria transmission season, can reduce parasite density and lead to a reduction of malaria episodes in the community.
Keywords:A, Alstonia boonei   ACT, artemisinin-based combination therapy   ALT, alanine aminotransferase   ANOVA, analysis of variance   AST, aspartate aminotransferase   BUN, blood urea nitrogen   CREA, creatinine   CE-FAO, European Commission-Food and Agriculture Organization   GGT, gamma glutamyltranspeptidase   H&  E, haematoxylin and eosin   ITN, insecticide treated nets   IPT, intermittent preventive treatment   K, Khaya ivorensis   KA, combined extraction of the 2 plants   K+A, 1:1 mixture of single plant extraction   LD50, lethal dose causing 50% mortality of experimental animals   LLIN, long lasting insecticide treated nets   LSD, least significant difference   OECD, Organization for Economic Co-operation and Development   P.f, Plasmodium falciparum   PBS, phosphate-buffered saline   RBC, red blood cell   U.S. EPA., United States Environmental Protection Agency   WBC, white blood cell   WHO, World Health Organization
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