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In vitro Antisickling and Sub Chronic Toxicity Studies of the Ethanol Extract of Cnidoscolus aconitifolius in Wistar Rats

Mojisola C. Cyril-Olutayo*, Norah O. Akinola, Temitope A. Adeyemo, Idris A. Oyemitan, Ronald A. Bejide, Joseph M. Agbedahunsi

Objective: The current study investigated the mechanism(s) of action of the antisickling property of Cnidiscolus aconitifolius ethanol extract and determined its histopathological effects on the liver, kidney and spleen of Wistar rats.

Methods: The antisickling properties of C. aconitifolius (CA) ethanol extract was tested at different concentrations, using the inhibitory and reversal models. Its effect on the density and mean cell volume of Hb S red blood cells was evaluated using the discontinuous density gradient assay. Ciklavit^®, an antisickling nutritional supplement was employed as positive while phosphate buffered saline (pH 7.0) was the negative control. Changes in percentage Red Cell Count (RBC) and Mean Cell Volume (MCV) in each density fraction were determined. Quantification of extracellular potassium leakage of RBC before and after incubation with CA extract was also conducted. CA was evaluated for its acute toxicity (LD₅₀) while sub-acute toxicity was evaluated by oral administration to Wistar rats for 28 days at 125, 250 and 500 mg/kg body weight. Rats were anaesthetized on the 29th day and blood collected through cardiac puncture for hematological analysis. The liver, kidney and spleen of the rats were harvested for histopathological examinations.

Results: The results showed that the percentage of dense cells in blood treated with CA was reduced significantly (p<0.05) compared to positive control. CA increased MCV of cells and inhibited basal potassium efflux with a rightward shift in the dose-response curve. The median lethal dose (LD₅₀) of CA was 3808 mg/kg; neither mortality nor structural or histological alterations were observed in the liver, kidney and spleen at the administered dose. The extract of CA enhanced erythropoiesis.

Conclusion: C. aconitifolius extract prevented potassium efflux and reduced red cell density in vitro. The CA crude extract was safe for consumption and increased the production of red blood cells in vivo.