EFFECT OF AQUEOUS ETHANOL LEAF EXTRACT OF COMBRETUM MICRANTHUM G. DON (COMBRETACEAE) ON SOME INDUCED SYSTEMIC INFLAMMATORY IMMUNE RESPONSE SYNDROMES IN MICE AND RATS
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Abstract
The cellular immunity is often suppressed by infection, disease conditions and drug medication that are in continous association with mankind. Combretum micranthum is a plant known for its antioxidant, antimicrobial, anti-inflammatory and various other properties of immune boosting activity. The Plant extract was assessed for its ability to protect from tissue destruction. The aqueous ethanol leaf extract was screened for previously reported phytoconstituents; Lorke,s method of 1983 was used to determined the LD50 in mice and rats using both i.p. and oral routes. Effect of the extract on haematological parameters were assessed pre- Cyclophosphamide (200 mg/kg i.p) induction on the 10th day and three days post induction according to Thatte et al. (1987) in four groups (n=5) of wistar rats. Group I received (1 ml/kg distilled water) while group II, III and IV received (50, 100 and 200 mg/kg of extract) respectively. The protective effect of the extract was also performed according to Pallable et al. (1998) by Escherichia. coli (1 ml of 2.5 × 108 Cfu) induction i.p. after 15th day pretreatment with the extract. Effect of the extract on Sheep red blood cell (SRBC) induced delayed type hypersensitivity reaction was evaluated according to Chang et al. (1998) on five groups (n=5) wistar rats. Group I received (1 ml/kg distilled water), group II, III and IV received 50, 100 and 200 mg/kg of extract while group V received 0.2 mg/kg dexamethasone. All the rats were immunized on the 7th day with 0.25 ml SRBC i.p. and the treatment continued to the 14th after which the challenging dose of 0.25 ml SRBC was administered via the plantar side of the right hind foot paw. Measurement were taken before the challenging dose (0 hr reading) and repeated at 3 and 24 hours later. Agar plate dilution method according to European Committee on Antimicrobial Susceptibility Testing (EUCAST), 2011 was used to test the susceptibility of Escherichia. coli, Staphylococcus. aureus, Klebsiella. pneumoneae, Salmonella. typhi vi and Proteus, mirabilis. The zone of inhibition compared with standard antibiotics was interpreted using interpretation criteria of EUCAST, 2011. The acute toxicity test showed an LD50 of 3807.9 mg/kg p.o. and 2154.1 mg/kg i.p. in mice; and > 5,000 mg/kg p.o. and 3807.9 mg/kg i.p. in rats. The phytochemical constituents revealed the presence of alkaloids, anthraquinones, carbohydrates, cardiac glycosides, flavonoids, saponins, steroids, tannins and terpenoids. The study showed dose dependent increase in WBC and neutrophil of the extract pretreated groups compared with the control group. Cyclophosphamide significantly reduced the WBC, RBC, HCT, HGB and ANC both in control and treated group. The extract protected all the pretreated mice up to 72 hr and beyond. The oedema at 24 hours was significantly higher in extract pretreated groups with 100 mg/kg having the highest response (p<0.001) that increased progressively to 24 hours. The extract was susceptible to all the tested organisms. Consequent upon these activities above, it can then be concluded that aqueous ethanol leaf extract of C. micranthum contains biological potent active principle which may be beneficial in cellular mediated immune disease conditions.
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