Acetylcholinesterase inhibitor from Anthocleista vogelii leaf extracts.

BACKGROUND: Anthocleista vogelii (Planch) is used in folk medicine in conditions related to inflammation and oxidative stress. This suggests that some of its constituents could possess ability to reduce oxidative damage to cells and ameliorate inflammatory conditions which are two prominent features in the brain of a person suffering from Alzheimer's disease. AIM OF STUDY: The leaf extracts were investigated for their bioactive constituents in our quest for novel compounds for the management of Alzheimer's disease. MATERIALS AND METHODS: Anthocleista vogelii crude leaf extract obtained from 80% methanol was successively partitioned with n-hexane, dichloromethane (DCM), ethyl acetate (EtOAc) and n-butanol. This gave four solvent fractions of the crude extract. These fractions and isolated compound were subjected to acetylcholinesterase (AChE) inhibitory activity test using the microplate assay method. The structure of the isolated compound was determined using spectroscopic methods (1D and 2D NMR, and MS). RESULTS: The results of the acetylcholinesterase inhibitory assay revealed that n-BuOH and ethyl acetate fractions had the best activity with IC50 of 564.58 and 727.63 µg/mL respectively. Activity directed fractionation of each of these fractions led to the isolation of the same compound identified as a C-glucosylflavone; isovitexin-7-O-methyl ether (swertisin). This rare C-glucosylflavone co-exist as two rotamers and exhibited duplication of NMR signals. It is reported from this plant for the first time. Swertisin displayed better acetylcholinesterase inhibitory activity with IC50 of 32.09 µg/mL than physostigmine (eserine) with IC50 of 56.09 µg/mL used as a standard. CONCLUSIONS: This study reports isolation and characterization of a rare C-glycosylflavone; isovitexin-7-O-methyl ether (swertisin). Isolated compound exhibited duplication of NMR signals and better acetylcholinesterase inhibitory activity than eserine used as a standard. The activity exhibited by this compound suggests that it could be useful in the management of Alzheimer's disease.

ISOLATION AND CHARACTERIZATION OF CHEMICAL CONSTITUENTS FROM CHRYSOPHYLLUM ALBIDUM G. DON-HOLL. STEM-BARK EXTRACTS AND THEIR ANTIOXIDANT AND ANTIBACTERIAL PROPERTIES.

BACKGROUND: The plant, Chrysophyllum albidum is indigenous to Nigeria and its stem-bark has wide application in traditional medicine for the treatment of infections and oxidative stress related diseases. The aim of the study was to isolate the chemical constituents responsible for the antioxidant and antibacterial activity from the stem-bark of the plant in order to justify some of its ethnomedicinal uses. MATERIALS AND METHODS: Crude extract of stem-bark of Chrysophyllum albidum obtained from 80% ethanol was successively partitioned with ethyl acetate (EtOAc) and n-butanol. The solvent fractions and isolated compounds were tested for antioxidant property using 2-2-diphenyl-1-picrylhydrazyl. Antibacterial activities were also assessed by means of agar-diffusion and broth micro dilution methods. EtOAc fraction was repeatedly fractionated on column chromatography to afford four compounds and their chemical structures were established using NMR (1D and 2D) and MS. RESULTS: Chromatographic fractionation of EtOAc fraction with the highest antioxidant and antimicrobial activities afforded stigmasterol (1),: epicatechin (2),: epigallocatechin (3): and procyanidin B5 (4).: Procyanidin B5 isolated for the first time from genus Chrysophyllum demonstrated the highest antioxidant activity with IC50 values of 8.8 µM and 11.20 µM in DPPH and nitric oxide assays respectively and equally demonstrated the highest inhibitory activity against Escherichia coli (MIC 156.25 µg/mL), Staphylococcus aureus (MIC 156.25 µg/mL), Pseudomonas aeruginosa (MIC 625 µg/mL) and Bacillus subtilis (MIC 156.25 µg/mL). CONCLUSION: The antibacterial and antioxidant activities of epicatechin, epigallocatechin and procyanidin B5 isolated from Chrysophyllum albidum stem-bark validate the folkloric uses.

A new bioactive thiophenolic glycoside from the leaf of Massularia acuminata (g. Don Bullock) ex Hoyle (Rubiaceae).

BACKGROUND: Massularia acuminata is a small tree or shrub of tropical rainforest. The leaves are used in Nigerian ethno-medicine for the treatment of microbial infections and pharmacological report suggested the leaf extract as possessing antioxidant activity. This study was therefore carried out to determine the most antioxidant and antimicrobial active fraction(s) of Massularia acuminata leaf and the constituent(s) responsible for the activities. MATHERIALS AND METHODS: The leaf of Massularia acuminata was investigated for in vitro antioxidant and antimicrobial activities, using a 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay and agar dilution method respectively. RESULTS: The ethyl acetate fraction demonstrated the best activities among the partitioned fractions tested. Bioassay guided purification of the most active ethyl acetate fraction led to isolation of a new thiophenolic glycoside, characterized as 4-(3',3'-dihydroxy-1-mercaptopropyl)phenyl glycosylpyranoside. CONCLUSION: The isolated compound from the leaf of Massularia acuminata demonstrated antioxidant and antimicrobial activities and may be responsible for the activities of leaf extract and its ethyl acetate fraction, hence this may justify its ethnomedicinal use.

Histomorphological and morphometric studies of the pancreatic islet cells of diabetic rats treated with extracts of Annona muricata.

Microanatomical changes in the pancreatic islet cells of streptozotocin induced diabetic Wistar rats were studied after treatment with methanolic extracts of Annona muricata leaves. Thirty adult Wistar rats were randomly assigned into three groups (control, untreated diabetic group, and A. muricata-treated diabetic group) of ten rats each. Diabetes mellitus was experimentally induced in groups B and C by a single intra-peritoneal injection of 80 mg/kg streptozotocin dissolved in 0.1 M citrate buffer. The control rats were intraperitoneally injected with an equivalent volume of citrate buffer. Daily intra peritoneal injections of 100 mg/kg A. muricata were administered to group C rats for two weeks. Post sacrifice the pancreases of the rats were excised and fixed in Bouin's fluid. The tissues were processed for paraffin embedding and sections of 5 mum thickness were produced and stained with H & E, Gomori aldehyde fuchsin, and chrome alum haematoxylin-phloxine for demonstration of the beta-cells of islets of pancreatic islets. Histomorphological and morphometric examination of the stained pancreatic sections showed a significant increase in the number, diameter, and volume of the beta-cells of pancreatic islets of the A. muricata-treated group (5.67 +/- 0.184 N/1000 mum(2), 5.38 +/- 0.093 mum and 85.12 +/- 4.24 mum(3), respectively) when compared to that of the untreated diabetic group of rats (2.85 +/- 0.361 N/1000 mum(2), 2.85 +/- 0.362 mum and 69.56 +/- 5.216 mum(3), respectively). The results revealed regeneration of the beta-cells of islets of pancreatic islet of rats treated with extract of A. muricata.

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.

Antibacterial activity of Piliostigma thonningii stem bark.

Piliostigma thonningii stem bark 60% methanolic extract exhibited activity against six out of eight bacterial isolates at a concentration of 20 mg/ml.

Preliminary evaluation of cytotoxic properties of Raphia hookeri fruit mesocarp.

The aqueous and n-butanol fractions of the methanolic extract of Raphia hookeri fruit mesocarp was evaluated for cytotoxic properties using three different bioassays. The n-butanol fraction exhibited greater cytotoxicity than the aqueous fraction when tested against brine shrimp, mosquito larvae, and 5-day old tadpoles, the cytotoxicity towards the tadpoles being the most pronounced (LC(50)=0.83 ppm).

Antibacterial activity of Nicotiana tabacum leaves.

Nicotiana tabacum leaf 60% methanolic extract exhibited antibacterial activity against six out of nine bacterial isolates at a concentration of 25 mg/ml.