Antisickling Studies On Three Nigerian Medicinal Plants
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Date
2015
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Abstract
The study evaluated the antisickling properties of three plants: Telfairia occidentalis Hook, F. family Cucurbitaceae (TO), Cnidoscolus aconitifolius (Mill.) I.M. Johnst. family Euphorbiaceae (CA) and Moringa oleifera Lam. family Moringaceae (MO) individually and in combination. It also determined the most active chemical constituent of the combination. This was with a view to providing information on the antisickling properties of the three plants and their combination.
The oven dried leaves were separately extracted by Soxhlet method using different solvents and maceration in 100 % ethanol. The individual plant extracts (TO, MO and CA) and various combinations (TMC, T2MC, TM2C and TMC2) were subjected to inhibitory and reversal antisickling assays. The T2MC, TO, MO and CA were subjected to different bioactivity test models such as red cell membrane stability, red cell density fractionation, HbSS polymerization and antioxidant assays to determine their effects on HbSS blood cells. The T2MC was then evaluated for acute toxicity in rats and furthermore fractionated into n-hexane, dichloromethane, ethyl acetate and methanol using vacuum liquid chromatography. Bioactivity directed studies were carried out on all the fractions to determine the putative constituents. Different chromatographic techniques: column chromatography, preparative thin-layer chromatography, flash chromatography and HPLC were used for further purification and isolation of the active constituents. Proton, 13Carbon NMR and GC/MS were used for the characterization of the isolated compounds. The results were subjected to statistical analysis using ANOVA at a p-value of 0.05.
The results showed that the cold ethanolic extract of each of TO, MO and CA have antisickling activities. The MO gave a significantly higher inhibitory activity (95.61 ± 0.47 %) while TO gave a significantly higher reversal activity (95.35 ± 0.81 %) at p < 0.05 than others. However, T2MC gave reversal (95.60 ± 0.28 %) and inhibitory (95.40 ± 0.15 %) activities which were significantly (p < 0.05) higher than the activities of Ciklavit® (76.88 ± 0.55 % inhibition and 64.20 ± 1.65 % reversal), respectively. The rate of decrease in polymerization (RDP) was directly proportional to concentration for TO, MO, CA and T2MC. However, T2MC gave 100 % RDP observed from 10 to 30 minutes at 4 mg/ml. The TO, MO, CA and T2MC showed dose dependent antioxidant and free radical scavenging activities but significantly (p < 0.05) low inhibition of nitric oxide. The membrane stability activities of the extracts increased with increased concentration with TO and T2MC giving significantly (p < 0.05) higher activities than Ibuprofen® control. All the extracts significantly (p < 0.05) reduced the percentage of the most dense cells (F4) after incubation. Four compounds: methyl 9-cis 11- trans-octadecadienoate; 9, 12-octadecadienoic acid (Linoleic acid); 1, 4- benzenedicarboxylic acid and hexadecanoic acid, methyl ester (Palmitic acid) were identified from the methanolic fraction of TO using GC/MS while two pure compounds: kaempferol-3-rhamnosyl-1→4-rhamnosyl and kaempferol-3-rhamnosyl-1→4-glucoside were isolated from T2MC by HPLC. The two isolated pure compounds were the active antisickling agents. This is the first report of the antisickling properties of the isolated pure compounds kaempferol-3-rhamnosyl-1→4-rhamnosyl (79.50 ± 0.445 % inhibition, 85.85 ± 3.471 % reversal) and kaempferol-3-rhamnosyl-1→4-glucoside (68.47 ± 4.243 % inhibition, 78.52 ± 3.217 % reversal).
The study concluded that the T2MC is a good candidate therapy for the treatment of sickle cell anaemia because it was not acutely toxic and had high antisickling properties
Description
xii, 270p
Keywords
antisickling, Nigerian medicinal, plants, Telfairia occidentalis, Cnidoscolus aconitifolius, Cucurbitaceae, Moringa oleifera