Pharmacological properties of the extract and some isolated compounds of Clausena lansium stem bark: anti-trichomonal, antidiabetic, anti-inflammatory, hepatoprotective and antioxidant effects.

ETHNOPHARMACOLOGICAL RELEVANCE: Clausena lansium (Fool's Curry Leaf) is used for various ethnomedical conditions in some countries, including bronchitis, malaria, viral hepatitis, acute and chronic gastro-intestinal inflammation, and as a spicy substitute of the popular Curry leaf tree (Murraya koenigii). AIM OF THE STUDY: This study was to evaluate the ethnomedical uses of the stem bark in inflammatory conditions, hepatotoxicity and to determine the anti-diabetic and anti-trichomonal properties of the plant. MATERIALS AND METHOD: Anti-trichomonal, in vivo and in vitro antidiabetic and insulin stimulating, anti-inflammatory, hepatoprotective and anti-oxidant activities using Trichomonas gallinae, glucose loaded rats and in vitro insulin secreting cell line (INS-1 cell), carrageenin-induced rat paw oedema, CCl(4)-induced hepatotoxicity and DPPH scavenging ability methods respectively for the extracts and some isolates were determined. RESULTS: A dichloromethane extract was superior over methanolic extract with respect to an anti-trichomonal activity which was measured after 24 and 48 h. The isolated compounds imperatorin and 3-formylcarbazole had the main anti-trichomonal activity (LC(50)s of 6.0, 3.0 and 3.6, 9.7 microg/mL after 24 and 48 h, respectively). Methanolic extract (100 mg/kg) induced maximum and significant (p<0.05) anti-hyperglycaemic activity of 15.8% at 30 min and a 38.5% increase in plasma insulin at 60 min, compared to control. The increase in plasma insulin after 60 min, compared to 0 min, was 62.0% (p<0.05). The significant 174.6% increase of insulin release from INS-1 cells (in vitro) at 0.1 mg/ml indicates that it mediates its antidiabetic action mainly by stimulating insulin release. Imperatorin and chalepin were the major active constituents increasing in vitro insulin release to 170.3 and 137.9%, respectively. 100 mg/kg of the methanolic extract produced an anti-inflammatory activity after 4 h. A sedative effect was not observed. 100 and 200 mg/kg of methanolic extract administered i.p., reduced CCl4-induced hepatotoxicity firstly by 5.3 and 8.4% reduction in phenobarbitone-sleeping time respectively, secondly by reversing the reduction in serum liver proteins by 7.0-8.8%, serum AST, ALT and ALP activities by 27.7-107.9% and thirdly by diminishing increased values of plasma AST, ALT and ALP activities by 13.2-83.8%. The extract exhibited antioxidant activities. CONCLUSION: The hepatoprotective activity of C. lansium is partly due to its anti-oxidant and anti-inflammatory properties and confirms its folkloric use in the treatment of gastro-intestinal inflammation, bronchitis and hepatitis. In addition the use of C. lansium stem bark would be useful in diabetes and trichomoniasis.

Hypoglycaemic constituents of Stachytarpheta cayennensis leaf.

The aqueous infusion (tea) of Stachytarpheta cayennensis leaves is used ethnomedically in Peru, Nigeria and other tropical countries for the management of diabetes. Oral administration (p. o.) of aqueous (125 mg/kg) and methanolic (2000 mg/kg) extracts of the leaves to alloxan-diabetic rats showed significant blood glucose reductions by 43 and 53%, respectively, at the end of a 4 hour period similar to the strong effect of glibenclamide (5 mg/kg, P. O.). The methanolic extract was successively partitioned into ethyl acetate, butanol and water fractions, and the same test showed that the butanol fraction (2000 mg/kg) had the highest (50%) hypoglycaemic activity at 4 hours after oral administration. It was also the most active fraction when tested in vitro [insulin release from an insulin secreting cell line (INS-1)] and was also active in normal rats and rats made hyperglycaemic by a glucose load. Its activity was comparable to that of glibenclamide (positive control) in these models. This active butanol fraction was subjected to chromatographic subfractionation; some subfractions reduced hyperglycaemia in alloxan-diabetic rats to 60 and 78% and induced insulin release from the INS-1 cells; other subfractions, however, gave hyperglycaemic activities IN VIVO and inhibition of insulin release from the INS-1 cells. Three major compounds of the butanol fraction were isolated and characterised as 6beta-hydroxyipolamide, ipolamide and isoverbascoside; they increased insulin secretion from INS-1 cells to 125, 128 and 127%, respectively, whereas glibenclamide increased insulin secretion to 157%. The results justify the ethnomedical use of the plant in the management of diabetes and suggest that the butanol fraction and some of its isolated constituents mediate their actions primarily by stimulating insulin release directly.

Antimicrobial and Antioxidant Activities of Some Nigerian Medicinal Plants

Ten Nigerian plants suggested from their ethnomedical uses to possess antimicrobial and antioxidant activities were studied for their anti-microbial and anti-oxidant properties. Antimicrobial activity was tested against Escherichia coli NCTC 10418, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus subtilis, Candida albicans, Candida pseudotropicalis and Trichophyton rubrum (clinical isolate). Trichilia heudelotti leaf extract showed both antibacterial and antifungal activities and was the most active against all the strains of bacteria tested. Boerhavia diffusa, Markhamia tomentosa and T. heudelotti leaf extracts inhibited the gram negative bacteria E.coli and P. aeruginosa strains whereas those of M. tomentosa, T. heudelotti and Sphenoceutrum jollyamum root inhibited at least one of the fungi tested. At a concentration of 312 µg/ml, hexane and chloroform fractions of T. heudelotti extract inhibited 6 and 14% of the fifty mult-idrug resistant bacteria isolates from clinical infectins, respectively. At ≤ 5mg/ml, the CHCl3 (64%) and aqueous (22%) fractions of T. heudelotti and those of CHCl3 (34%) and EtOAC (48%) of M. tomentosa gave the highest inhibition that was stronger than their corresponding methanol extracts. The corresponding EC50 of the extracts on M. acuminata, T. heudelotti, E. senegalensis and M. tomentosa were 4.00, 6.50, 13.33, and 16.50 ig/ml using the TLC staining and 1,1-dipheyl-2-picry-hydrazyl (DPPH) free radical scavenging assay. Therefore, leaf extracts of M. tomentosa and T. heudelotti, especially the latter, possess strong antimicrobial and antioxidant activities and should be further investigated. These activities justified the ethnomedical uses of these plants

Anti-trichomonal, biochemical and toxicological activities of methanolic extract and some carbazole alkaloids isolated from the leaves of Murraya koenigii growing in Nigeria.

The methanolic extract of Murraya koenigii leaf was screened for toxicological and biochemical effects on rats because of the folkloric uses as an anti-dysentery and anti-diabetes. The extract was moderately toxic (LD(50)=316.23 mg/kg body weight) to rats and had appreciable effect on the liver and kidney at higher doses leading to liver inflammation. It had little or no effect on haematology and relative organ weight of lungs, heart and spleen. Acute doses (500 mg/kg) reduced significantly serum globulin, albumin, urea, glucose, total protein, aspartate transaminase (AST), and increased cholesterol and alanine transaminase (ALT) indicating hepatic injury. However, chronic administration for 14 days gave a significant (p<0.05) reduction in the serum cholesterol, glucose, urea, bilirubin, ALT and AST showing that the plant has hypoglycaemic and hepatoprotective effects after prolonged use. The activity demonstrated by some of the isolated carbazole alkaloids and their derivatives against Trichomonas gallinae confirmed that the anti-trichomonal activity of the leaf may be due to its carbazole alkaloids. The order of activity was C(18)>C(23)>C(13). Girinimbine and girinimbilol with IC(50) values of 1.08 and 1.20 microg/ml were the most active. Acetylation of girinimbilol and mahanimbilol improved their activities to 0.60 and 1.08 microg/ml.

Antimicrobial and antioxidant activities of some Nigerian medicinal plants.

Ten Nigerian plants suggested from their ethnomedical uses to possess antimicrobial and antioxidant activities were studied for their anti-microbial and anti-oxidant properties. Antimicrobial activity was tested against Escherichia coli NCTC 10418, Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus subtilis, Candida albicans, Candida pseudotropicalis and Trichophyton rubrum (clinical isolate). Trichilia heudelotti leaf extract showed both antibacterial and antifungal activities and was the most active against all the strains of bacteria tested. Boerhavia diffusa, Markhamia tomentosa and T. heudelotti leaf extracts inhibited the gram negative bacteria E. coli and P. aeruginosa strains whereas those of M. tomentosa, T. heudelotti and Sphenoceutrum jollyamum root inhibited at least one of the fungi tested. At a concentration of 312 microg/ml, hexane and chloroform fractions of T. heudelotti extract inhibited 6 and 14% of the fifty multi-drug resistant bacteria isolates from clinical infections, respectively. At < or = 5 mg/ml, the CHCl(3) (64%) and aqueous (22%) fractions of T. heudelotti and those of CHCl(3) (34%) and EtOAC (48%) of M. tomentosa gave the highest inhibition that was stronger than their corresponding methanol extracts. The corresponding EC(50) of the extracts on M. acuminata, T. heudelotti, E. senegalensis and M. tomentosa were 4.00, 6.50, 13.33, and 16.50 ig/ml using the TLC staining and 1,1-dipheyl-2-picry-hydrazyl (DPPH) free radical scavenging assay. Therefore, leaf extracts of M. tomentosa and T. heudelotti, especially the latter, possess strong antimicrobial and antioxidant activities and should be further investigated. These activities justified the ethnomedical uses of these plants.

Ethno-veterinary medicine: screening of Nigerian medicinal plants for trypanocidal properties.

Trypanosoma congolense and T. brucei bloodstream form parasites were propagated axenically in suitable standard media at 34 degrees C. The effects of 33 plant extracts, fractions and pure compounds were evaluated on two clones of T. brucei and drug-sensitive and multi-drug-resistant clones of T. congolense. The cytotoxic activity of the trypanocidal extracts was also evaluated on calf aorta endothelial cells in vitro. Of the extracts tested, 22% killed T. congolense IL 1180 at a concentration of 100 microg/ml while 18% killed 90-100% of T. brucei ILTat 1.4 at the same concentration. However, 6% of the active extracts killed 93% of a dyskinetoplastid form of T. brucei IL Tat 1.1, indicating that the intact kinetoplast is a target of some of the compounds tested. Of the 12 extracts that displayed activity against drug sensitive trypanosomes, 66.7% had trypanocidal activity on a multi-drug-resistant clone, T. congolense IL 3338. The extracts of Eugenia uniflora, Acacia artaxacantha, Terminalia ivorensis, T. superba and Alchornea cordifolia had median lethal concentrations of between 13 and 69 microg/ml on both the drug-sensitive, IL 1180 and multi-drug-resistant clone, IL 3338. The median lethal doses of the active plant extracts on the calf aorta endothelial cells varied between 112 and 13750 microg/ml while the calculated selective indices ranged between 0.71 and 246.8 indicating bright prospects for the development of some of these extracts as potential trypanocidal agents.

Minor furocoumarins of Murraya koenigii.

Xanthotoxin, isobyakangelicol, phellopterin, gosferol, neobyakangelicol, byakangelicol, byakangelicin and isogosferol are reported as minor furocoumarins of Murraya koenigii seeds.