Integrating in vitro and in silico experimental validation to elucidate the antisickling properties of sub-fractions from aqueous Khaya senegalensis and Diospyros mespiliformis Stem barks extracts

Abstract

Sickle cell disease (SCD) is the most common inherited haematological disorder in the world, with Nigeria bearing the highest burden in Africa. Diospyros mespiliformis (DM) and Khaya senegalensis (KS) are plants with acclaimed antisickling potentials but without scientific backings. This study investigated the in vitro, in silico and toxicological profiles of chromatographic sub-fractions of DM and KS stem barks (SBDm and SBKs). Antisickling activities were assessed using standard red blood cells sickling reversal, haemoglobin polymerization inhibition and osmotic fragility tests. Toxicological evaluation involved oral administration of the most active sub-fractions to rats for four (4) weeks, followed by assessment of lipid profile, haematological indices as well as liver and kidney function parameters. Data were analyzed using one-way ANOVA at p<0.05. Sub-fractions 3 and 4 of DM (F3SBDm and F4SBDm), and sub-fraction 4, 6 and 7 of KS (F4SBKs, F6SBKs and F7SBKs) showed the strongest antisickling effects. In silico analysis using GC-MS and LC-MS identified 1-(Isopropylamino)-3-(1-naphthylmethoxy)-2-propanol, and 1,5-Dimethyl-4-[(4-nitro-benzylidene)-amino]-2-phenyl-1,2-dihydro-pyrazol-3-one as the bioactive compounds with high binding affinity to Haematopoietic Cell Kinase, Calcium-ATPase, Extracellular signal regulated protein kinase and deoxyhemoglobin. F6SBKs significantly improved lipid profile by increasing HDL and reducing LDL cholesterol, while F4SBDm enhanced erythrocyte and haemoglobin levels. Both F6SBKs and F4SBDm reduced serum ALT, AST and ALP, indicating hepatoprotection. Histopathology showed normal liver an d kidney architecture after treatment with the sub-fractions from both SBDm and SBKs, except F3SBDm, which caused hepatic damage. The study concludes that F4SBDm and K6SBKs displayed good antisickling activities by reversing sickling of red blood cells and preventing haemoglobin polymerization, with no toxicity associated to both kidney and liver tissues.