RESUMEN
Aim: Onchocerciasis is an endemic parasitic disease in sub-Saharan Africa that significantly impacts animal and human health. In Northern Cameroon, medicinal plants from the Combretum genus are used for onchocerciasis traditional treatment although there is no scientific evidence of their antifilarial potential. This study evaluates the in vitro macro- and microfilaricidal properties of water extracts from Combretum nigricans in Onchocerca ochengi. Material and Methods. O. ochengi microfilariae and adult male worms were recovered from cowhide fragments. Oxidative stress indicators and motility tests were used to assess the filaricidal impact. Female albino rats were used to test for acute toxicity. The contents of secondary metabolites were quantified. Results: The bark aqueous extract was more active on macrofilariae at 1 mg/mL for 24 h (100%) than the leaf (63.9%) and root (75%) extracts at the same concentration. Likewise, a stronger microfilaricidal effect was found with this extract at 0.5 mg/mL for 1 h (100%) compared to root and leaf extracts. The dose-response effect with the bark extract gave an inhibitory concentration 50 (IC50) of 351 µg/mL vs. 113 µg/mL for flubendazole after 24 h incubation, while the microfilaricidal efficacy revealed an IC50 of 158.7 µg/mL vs. 54.09 µg/mL for ivermectin after one-hour incubation. Examining stress indicators on parasite homogenates showed that macrofilaricidal activity is associated with a significant increase in nitric oxide, glutathione, and malondialdehyde generation and a decrease in catalase activity. At 2000 mg/kg, rats showed no harm. The phytochemical investigation revealed that the barks contained more phenolic acids, condensed tannins, flavonoids, and saponins than the leaves (p < 0.001). Conclusion: These findings support C. nigricans' antifilarial activity and identify oxidative stress indicators as prospective treatment targets in O. ochengi. It would be interesting to conduct in vivo studies to understand their antifilarial activity better.
RESUMEN
PURPOSE: Onchocerciasis is a neglected tropical disease that remains endemic in sub-Saharan African countries. Unfortunately, only a few microfilaricidal agents have been approved so far. This study aimed to assess the in vitro macro and microfilaricidal potentialities of the hydro-methanolic extracts of the different powdery fractions of Khaya senegalensis against Onchocerca ochengi. METHODS: Adult male worms and microfilariae (mf) of O. ochengi were isolated from cowhides in Ngaoundere II, Cameroon. Parasites were incubated for 4 h (mf) or 48 h (adult worms) in RPMI-1640 medium in the presence or absence of ivermectin, flubendazole, or hydro-methanolic extracts of different plant powdery fractions obtained by controlled differential sieving. The filaricidal effect was evaluated using motility (mfs) and mortality tests (worms) and oxidative stress parameters. Cytotoxicity and acute toxicity tests were performed on monkey-derived kidney cell lines (LLC-MK2) and Swiss albino mice, respectively, and selectivity indexes were determined. Phytochemical screening was also carried out using high-performance liquid chromatography/UV (HPLC/UV), molecular networking, and through quantification of phenolic contents. RESULTS: The hydro-methanolic extracts of 0-63 µm fractions from leaves and barks exhibited the strongest macrofilaricidal activities with lethal concentrations 50 of 162.4 and 208.8 µg/mL respectively versus 22.78 µg/mL for flubendazole. These two fractions also showed the fastest microfilaricidal activities (T1/2 of 1 h), although it was low when compared to ivermectin (T1/2 < 1 h). Their macrofilaricidal effects were accompanied by a significant inhibition of nitric oxide secretion and a significant increase of glutathione and catalase activity compared to the untreated group. However, no effect was found on superoxide dismutase activity, the GABAergic and glutamatergic receptors. Although neither extract was toxic to Swiss mice until a dose of 2000 mg/kg body weight, the 0-63 µm leaf fraction hydro-methanolic extract was selectively more effective on worms than bark extract (SI = 1.28 versus 0.34). Both extracts were found to contain some flavonoids including procyanidin-, rutin-, myricetin-, and naringenin derivatives as well as new unknown compounds. However, the total polyphenol, flavonoid and tannin contents of the leaf extract were significantly greater (P < 0.05) than that of the bark extract. CONCLUSION: These results support the anti-filarial effect of K. senegalensis leaves and highlight stress oxidative markers as new therapeutic targets in O. ochengi. Further, in vivo experiments are required in understanding their anti-parasitic properties, and testing combinations of fine fractions.