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Nutritive value and haemolytic properties (in vitro) of the leaves of Vernonia amygdalina on human erythrocyte.

BACKGROUND: Vernonia amygdalina (Bitter leaf) is popularly consumed in many homes in Nigeria as a vegetable in soup preparation, while the infusion is used in folk medicine in the treatment of malaria fever. In this study, fresh leaves of Vernonia amygdalina were subjected to abrasion (to remove the bitter taste) as is normally done in soup preparation, subsequently the nutrient, anti-nutrient and zinc bioavailability of the processed and unprocessed leaf were determined. The heamolytic effect of the leaf infusion on human erythrocytes (Genotype-SS, AS, AA) was also determined. RESULT: The study revealed that Vernonia amygdalina leaf has high protein (33.3%), fat (10.1%), crude fibre (29.2%), ash (11.7%), mineral (Na, K, Ca, Mg, Zn & Fe), phytate (1015.4mg/100g) and tannin (0.6%) content, while it contain low cyanide (1.1mg/kg). Abrasion of the leaf caused a significant decrease (P<0.05) in both the nutrient and antinutrient content of the leaf except in carbohydrate where there was a significant increase (P<0.05) and Mg where there was no significant change (P>0.05). The result of the calculated [Ca][phytate]/ [Zn] molar ratio (the best index for the prediction of Zn bioavailability) indicated that the phytate content of both the processed and unprocessed Vernonia amygdalina leaf will not reduce the Zn content to a critical level (0.5mol./kg). Further studies on the leaf revealed that the infusion induced the haemolysis of human erythrocyte (In vitro). Human genotype-SS (1024) were highly susceptible to haemolysis induced by Vernonia amygdalina infusion, genotype-AS moderately susceptible (512) while genotype-AA (256) is highly resistant to haemolysis induced by the same infusion. CONCLUSION: Bitter leaf has high nutrient and antinutrient content, however, abrasion will bring about a significant decrease (P<0.05) in both. The infusion of the Vernonia amygdalina leaf induces the haemolysis of mammalian erythrocyte in vitro, with Human-SS having the highest susceptibility.

Biological Availability↗

Hypoglycaemic and antidiabetic activity of acetonic extract of Vernonia colorata leaves in normoglycaemic and alloxan-induced diabetic rats.

The aqueous extract of Vernonia colorata (Willd.) Drake (Composeae) leaves is used by African traditional medicine practitioners as a remedy for the treatment of diabetes. Our previous studies have shown the hypoglycaemic activity of the aqueous extract of Vernonia colorata leaves (300 mg/kg, per os) in normoglycaemic rats. The aim of the present study was to investigate the hypoglycaemic and antidiabetic activity of acetonic and hexanic extracts of the leaves of Vernonia colorata in order to further discriminate the type of extract which provides a better antidiabetic activity. Experiments were performed in normoglycaemic and alloxan-induced diabetic rats. The acetonic extract of the leaves of Vernonia colorata (AELVC) (100 mg/kg, per os) induced a significant decrease of blood glucose in normoglycaemic rats. The glycaemia varied from 4.72+/-0.11 to 3.72+/-0.22 mmol/l (p<0.05, n=5) 3 h after AELVC administration per os. In contrast, the hexanic extract of the leaves of Vernonia colorata (HELVC) increased significantly the glycaemia in normoglycaemic rats. Like glibenclamide, AELVC has an antihyperglycaemic effect in oral glucose tolerance test (OGTT) and alloxan-induced diabetic rats. These results have shown that: (i) AELVC and HELVC have an opposite effect on basal blood glucose in normoglycaemic rats, suggesting that the mechanisms of action of both above-mentioned extracts are different; (ii) AELVC has also an antidiabetic activity in hyperglycaemic rat models.

Acetone↗

Properties of a virus isolated from Vernonia amygdalina Del. in Lagos, Nigeria.

A previously uncharacterized virus tentatively named Vernonia green vein-banding virus (VGVBV) was isolated from Vernonia amygdalina Del. ("bitterleaf") from Lagos, Nigeria. The virus was mechanically transmissible but had a narrow host range restricted to Nicotiana benthamiana, Chenopodium quinoa and C. amaranticolor. It was also transmissible in a non-persistent manner by Myzus persicae. The virus was purified from N. benthamiana and about 750 nm long flexuous rod-shaped particles were observed in purified preparations as well as in leaf-dips of Vernonia sp. Inclusion bodies in the form of pinwheels and scrolls were observed in ultrathin sections of Vernonia leaves by electron microscopy. M(r), of the viral coat protein was estimated to be about 34 K. In indirect ELISA, all 20 samples from naturally infected Vernonia sp. reacted positively with a potyvirus-specific monoclonal antibody (MAb) as well as with an antiserum raised against VGVBV. Apart from the homologous antigen, the VGVBV antiserum reacted only with Plum poxvirus (PPV). The VGVBV reacted strongly with the antisera to Bean yellow mosaic virus (BYMV), Bean common mosaic virus (BCMV) and Amaranthus leaf mottle virus (AmLMV) but weakly with antisera to PPV and Cowpea aphid-borne mosaic virus (CABMV) (all members of the family Potyviridae, the genus Potyvirus) in at least one of the assays used (indirect ELISA, dot-blot immunoassay and Western blot analysis). The results of our host range, cytopathological and serological studies and the available literature indicate that a hitherto difficult to transmit VGVBV has only been reported from Nigeria. We consider VGVBV a candidate for a new potyvirus. This virus should be further investigated to collect sufficient data for a qualified proposal of VGVBV as a new potyvirus.

Animals↗

Novel cationic amphiphilic derivatives from vernonia oil: synthesis and self-aggregation into bilayer vesicles, nanoparticles, and DNA complexants.

Self-assembling nanostructures were prepared from novel cationic amphiphilic compounds synthesized from vernonia oil, a natural epoxydized triglyceride. The presence of a 12,13-epoxy group on the C18 unsaturated fatty acid, vernolic acid, which is the main constituent of vernonia oil, permitted the synthesis of novel amphiphilic derivatives with a hydrogen-bonding hydroxyl and a cationic headgroup moiety on adjacent carbon atoms. The amphiphiles were prepared in a two-stage synthesis that comprised opening of the epoxy groups with a haloacetic acid, followed by quaternization of the halo group with a tertiary amine containing a C12 aliphatic chain. Intact vernonia oil as the starting material gave a triple-headed cationic amphiphile, containing three vernolic acid derived moieties connected through a glycerol backbone. A single-headed amphiphile with two alkyl chains and a single quaternary ammonium headgroup was synthesized from the methyl ester of vernolic acid as the starting material. The triple-headed derivative could form nonencapsulating structures. Cholesterol was required in the formulation (1:1) to make spherical vesicles that could encapsulate a water-soluble marker. The single-headed derivative, however, formed spherical encapsulating vesicles without cholesterol. TEM, NMR, and FT-IR were used to characterize the vesicles, and molecular structure vs morphology relationships were postulated on the basis of these data. The triple-headed amphiphile also formed a DNA complex that was highly resistant to hydrolysis by DNase. This amphiphile-DNA complex was used as vector for gene transfer in cell culture demonstrating efficient DNA transfection.

Animals↗

Effect of the crude extract of Vernonia polyanthes Less. on blood pressure and renal sodium excretion in unanesthetized rats.

This study evaluated the effect of oral crude Vernonia polyanthes Less. hydroalcoholic extract administration (CHE, 0.5 and 1.0 g/kg body wt., daily for 7 days) on arterial blood pressure and renal sodium excretion in conscious rats. CHE administration decreased arterial blood pressure dose-dependently followed by a significant rise in creatinine clearance and a fall in fractional post-proximal sodium excretion was compared to the control group. These results suggest that blood pressure decrease induced by the oral crude Vernonia hydroalcoholic extract may be blunted by reduction of the post-proximal renal sodium excretion. Thus, the present study shows that Vernonia extract is a potential vasodilatation agent in normotensive rats without any effects on renal tubule autoregulation mechanisms.

Animals↗

Characterization of copolymer hydroxybutyrate/hydroxyvalerate from saponified vernonia, soybean, and "spent" frying oils.

Poly(beta-hydroxyalkanoate)s (PHAs) were biosynthesized by Ralstonia eutropha (formerly known as Alcaligenes eutrophus) by using saponified soybean, vernonia, and "spent" frying oils. These PHAs were isolated and characterized by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS), gas chromatography/mass spectrometry (GC/MS), proton nuclear magnetic resonance spectrometry (1H NMR), and 2-dimensional homonuclear (1H-1H) correlation spectroscopy (COSY). The analytical results revealed that the PHAs produced from saponified vernonia and soybean oils were copolymers of hydroxybutyrate (HB) and hydroxyvalerate (HV), that is, P(HB/HV)s, whereas the saponified "spent" frying oil produced only poly(beta- hydroxybutyrate) (PHB) homopolymer. MALDI-MS, GC/MS, and NMR independently confirmed the composition of the PHAs. Saponified soybean oil and vernonia oil PHAs contained approximately 4 and 1% HV units, respectively. For comparison, commercial PHB and P(HB/HV), produced by R. eutropha by using glucose and a cosubstrate of glucose and propionic acid, respectively, as carbon sources, were similarly characterized.

Dietary Fats, Unsaturated↗

Fatty acid selectivity of a lipase purified from Vernonia galamensis seed.

Vernonia galamensis is an annual herb whose seed oil contains high levels of an epoxy fatty acid, vernolic (cis-12,13-epoxy cis-9-octadecenoic) acid. The seed also contains lipase activity in the dormant state. A lipase was purified from the seed and its substrate specificity studied in isooctane. The lipase shows pronounced selectivity for the native triacylglycerol, trivernolin. The rate of hydrolysis of triolein, the corresponding non epoxy triacylglycerol, is only 3% of that of trivernolin. In the acidolysis of tricaprylin using a mixture of fatty acids, the Vernonia lipase also showed selectivity for vernolic acid. Michaelis-Menten kinetics of the hydrolysis of triacylglycerols revealed that the observed high selectivity of the Vernonia lipase for trivernolin was mainly due to a higher Vmax for trivernolin. The Vmax value for the hydrolysis of trivernolin was 5 times higher than that for triolein. This novel substrate specificity is an adaptation by the seed lipase to the triacylglycerols of the seed oil that contain up to 80% vernolic acid.

Epoxy Compounds↗

Isolation, partial characterisation and immunomodulating activities of polysaccharides from Vernonia kotschyana Sch. Bip. ex Walp.

The roots from Vernonia kotschyana Sch. Bip. ex Walp. (Baccharoides adoensis var. kotschyana (Sch. Bip. ex Walp.) M.A. Isawumi, G.El-Ghazaly & B. Nordenstam) (Asteraceae) are used in Malian folk medicine for the treatment of gastritis, gastro duodenal ulcers, as an aid to ameliorate digestion and as a wound healing remedy. Since a common feature among these conditions is related to immune responses, immunomodulating activities of fractions isolated from both the 50 degrees C and the 100 degrees C water extracts from Vernonia kotschyana were investigated in this study. The active principles were identified as acidic polysaccharide fractions, containing pectic arabinogalactan type II structures, which showed both complement fixing ability and T-cell independent induction of B-cell proliferation in vitro. Some activity was also observed on macrophages. The present study may provide additional support for the popular use of this plant to improve intestinal health.

Animals↗

Bioactive sesquiterpene lactones from the leaves of Vernonia amygdalina.

Phytochemical analysis of the leaves of Vernonia amygdalina yielded two known sesquiterpene lactones: vernolide and vernodalol. The two compounds were tested by agar dilution method against 10 bacteria strains and 5 fungi species. Both compounds exhibited a significant bactericidal activity against five Gram positive bacteria while lacking efficacy against the Gram negative strains. In the antifungal test, while vernolides exhibited high activity with LC(50) values of 0.2, 0.3 and 0.4 mg/ml against Penicillium notatum, Aspergillus flavus, Aspergillus niger and Mucor hiemalis, respectively, vernodalol showed moderate inhibitions against Aspergillus flavus, Penicillium notatum and Aspergillus niger with LC(50) values of 0.3, 0.4 and 0.5 mg/ml, respectively. Both compounds were ineffective against Fusarium oxysporum, a microbe known to be highly resistant to chemical agents. However, the antimicrobial results of this study correspond positively with the claimed ethnomedical uses of the leaves of Vernonia amygdalina in the treatment of various infectious diseases.

Anti-Bacterial Agents↗

Ent-kaurane glycosides and sesquiterpene lactones of the hirsutinolide type from Vernonia triflosculosa.

Investigation of the aerial parts of Vernonia triflosculosa afforded three hirsutinolides of which 8alpha-(4alpha-hydroxymethacryloyloxy)-10alpha-hydroxy-1,13-dimethoxy-hirsutinolide is new, three ent-kaurane diterpenes, among which the 19-[alpha-L-arabinopyranosyl-(1-->2)-beta-D-glucopyranosyl] esters of 16beta-hydroxy-ent-kauran-19-oic acid and of 16beta,17-hydroxy-ent-kauran-19-oic acid are also new. Diterpenes are reported here for the first time in the large genus Vernonia. Their structures were elucidated using 1D and 2D NMR measurement as well as ESI, CIMS, and HRMS analysis. Two hirsutinolides were studied for their NF-kappaB DNA binding activity in HaCaT cells (a human cell line similar to keratinocytes) and for their inhibition on IL-8 production in HeLa cells.

Cells, Cultured↗

In vitro cytotoxic elemanolides from Vernonia lasiopus.

Two new elemanolides, epivernodalol and lasiopulide, were isolated after chromatographic separation of the alcoholic extract of the dried aerial parts of the Vernonia lasiopus. These elemanolides are new C-10 epimers of the sesquiterpene lactones vernodalol and demethylacroylated vernodalol isolated from other species of Vernonia. Both elemanolides showed in vitro cytotoxicity against human cancer cell lines in culture. This is the first report of isolation and cytotoxic activity of the two elemanolides from V. lasiopus.

Antineoplastic Agents, Phytogenic↗

[Studies on the constituents from the herb of Vernonia patula].

OBJECTIVE: To isolate and elucidate the chemical constituents of the whole plant of Vernonia patula, and to provide samples for biological activity screening. METHOD: The chromatography on silica gel was used and the structures were determined by IR, NMR and MS spectra analysis and physicochemical property comparion. RESULT: Four triterpenoids were isolated and identified as bauerenyl acetate(I), friedelin(II), epifriedelanol(III), 20(30)-taraxastene-3 beta, 21 alpha-diol(IV). CONCLUSION: Compounds I and IV were isolated from genus Vernonia for the first time and compounds II and III were isolated from the whole plant of V. patula for the first time. The four compounds show no inhibitory effect on the growing of PC-12 cells.

Animals↗

Separation of flavonoids from the seeds of Vernonia anthelmintica Willd by high-speed counter-current chromatography.

Several flavonoids including 2',3,4,4'-tetrahydroxychalcone, 5,6,7,4'-tetrahydroxyflavone and butin, were separated from the seeds of Vernonia anthelmintica Willd by high-speed counter-current chromatography using a two-step operation. Two different types of solvent systems were used: chloroform-dichloromethane-methanol-water (2:2:3:2, v/v) and 1,2 dichloroethane-methanol-acetonitrile-water (4:1.1:0.25:2, v/v). From 1 kg of seeds of Vernonia anthelmintica Willd the method yielded about 45 mg of 2',3,4,4'-tetrahydroxychalcone, 40mg of 5,6,7,4'-tetrahydroxyflavone, and 55 mg of butin. Each isolated component showed 95-97% purity as determined by high-performance liquid chromatography analysis. These purified compounds were characterized by MS and NMR.

Chromatography, High Pressure Liquid↗

Measurement of glomerular filtration rate using inulin prepared from Vernonia herbacea, a Brazilian native species.

Vernonia herbacea (Vell.) Rusby (Asteraceae) is a perennial herb native to the cerrado vegetation of tropical areas in Brazil, which accumulates inulin in the underground reserve organs. The aim of this paper was to determine whether the inulin extracted from V. herbacea could replace commercial inulin for the measurement of glomerular filtration rate (GFR). Underground organs of vegetative plants were collected from a preserved area of the Brazilian cerrado. The inulin fraction utilized was obtained by ethanol precipitation after discarding the high molecular mass fructans in the freeze-thawing precipitate. GFR was determined in male Wistar rats anesthetized with inactin (100 mg/kg), which received intravenously commercial inulin obtained from Dahlia sp (Sigma) or Vernonia herbacea inulin (30 mg/100 g) as a priming dose and 0.05 mg min-1 100 g-1 as a sustaining dose in isotonic saline at the rate of 0/055 ml/min. Clearance was determined during 3 periods, with urine collected from the bladder and blood from the carotid artery. There was no significant difference in the GFR measured by clearance in inulin from both sources even when the plasma concentration of inulin from V. herbacea was doubled. The mean arterial pressure did not vary after the application of both inulins, indicating that they do not produce systemic side effects. The filtered load and the excreted amount of inulin from V. herbacea were equal, showing that the substance is not influenced by tubular function. These results demonstrate that the inulin from V. herbacea can substitute for imported inulin for the determination of GFR and in experiments of kidney microperfusion as a marker of tubular water reabsorption.

Animals↗

Evaluation of antipyretic potential of Vernonia cinerea extract in rats.

The methanol extract of the whole plant of Vernonia cinerea (MEVC) was evaluated for its antipyretic potential on normal body temperature and yeast-induced pyrexia in rats. MEVC significantly reduced the normal body temperature at doses of 250 and 500 mg/kg body weight p.o. MEVC also lowered the elevated body temperature in the case of yeast-induced pyrexia in a dose dependent manner. The antipyretic effect of the extract at a dose of 500 mg/kg was identical to that of the standard drug paracetamol.

Administration, Oral↗

Diacylglycerol acyltransferases from Vernonia and Stokesia prefer substrates with vernolic acid.

Genetic engineering of common oil crops for industrially valuable epoxy FA production by expressing epoxygenase genes alone had limited success. Identifying other key genes responsible for the selective incorporation of epoxy FA into seed oil in natural high accumulators appears to be an important next step. We investigated the substrate preferences of acyl CoA:diacylglycerol acyltransferases (DGAT) of two natural high accumulators of vernolic acid, Vernonia galamensis and Stokesia laevis, as compared with a common oilseed crop soybean. Developing seed microsomes were fed with either [14C]oleoyl CoA or [14C] vernoloyl CoA in combinations with no exogenous DAG or with 1,2-dioleoyl-sn-glycerol, 1-palmitoyl-2-vernoloyl-sn-glycerol, 1,2-divernoloyl-sn-glycerol, 1,2-dioleoyl-rac-glycerol, or 1,2-divernoloyl-rac-glycerol to determine their relative incorporation into TAG. The results showed that in using sn-1,2-DAG, the highest DGAT activity was from the substrate combination of vernoloyl CoA with 1,2-divernoloyl-sn-glycerol, and the lowest was from vernoloyl CoA or oleoyl CoA with 1,2-dioleoyl-sn-glycerol in both V. galamensis and S. laevis. Soybean DGAT was more active with oleoyl CoA than vernoloyl CoA, and more active with 1,2-dioleoyl-sn-glycerol when oleoyl CoA was fed. DGAT assays without exogenous DAG, or with exogenous sn-1,2-DAG fed individually or simultaneously showed consistent results. In combinations with either oleoyl CoA or vernoloyl CoA, DGAT had much higher activity with rac-1,2-DAG than with their corresponding sn-1,2-DAG, and the substrate selectivity was diminished when rac-1,2-DAG were used instead of sn-1,2-DAG. These studies suggest that DGAT action might be an important step for selective incorporation of vernolic acid into TAG in V. galamensis and S. laevis.

Acetyl Coenzyme A↗

Structural and immunological studies of a pectin and a pectic arabinogalactan from Vernonia kotschyana Sch. Bip. ex Walp. (Asteraceae).

Two polysaccharides, a pectin (Vk100A2b) and a pectic arabinogalactan (Vk100A2a) with mean Mw 2 x 10(4) and 1.15 x 10(6)Da, respectively, were isolated from the dried powdered roots of Vernonia kotschyana Sch. Bip. ex Walp. by hot water extraction followed by fractionation on DEAE-Sepharose fast flow and Sephacryl S-400 HR. The pectin showed low-complement fixation activity and no influence on proliferation of B or T cells, while the pectic arabinogalactan showed a potent, dose-dependent complement fixation activity and a T cell independent induction of B-cell proliferation. Both polysaccharides induced chemotaxis of human macrophages, T cells and NK cells. exo-alpha-L-arabinofuranosidase and exo-beta-D-galactosidase digestion followed by component sugar and methylation analysis indicated that Vk100A2a consisted of a highly branched rhamnogalacturonan core with approximately 50% of the rhamnose 1,2,4-substituted, side chains rich in terminal-, 1,5-linked and 1,3,5-branched arabinose and terminal-, 1,4-, 1,6-linked and 1,3,6-branched galactose. The enzyme resistant part of Vk100A2a still showed strong complement fixating activity, suggesting that this activity may at least in part be expressed by carbohydrate structures present in the enzyme resistant, inner portion of the polymer.

Chemotaxis↗

Fructan metabolising enzymes in rhizophores of Vernonia herbacea upon excision of aerial organs.

The activities of fructan metabolising enzymes and fructan contents are reported for rhizophores of Vernonia herbacea (Vell.) Rusby induced to sprouting by shoot excision. The activities of fructan exohydrolase (1-FEH), sucrose: sucrose fructosyltransferase (1-SST), fructan: fructan fructosyltransferase (1-FFT) and invertase (INV) and the fructan contents were analysed every 3-4 days for 1 month by colorimetric and chromatographic methods. Sprouting of new shoots started on day 9. 1-FEH activity increased after day 13 and reached its maximum value 20 days after shoot excision. A gradual decrease in 1-SST activity was detected between days 3 and 9. 1-FFT activity exhibited fluctuations throughout the experimental period and a peak of activity for invertase was detected 9 days after shoot excision. Variation in fructan contents in vivo included a decrease until day 13 after which, levels remained practically unchanged. Fructan depolymerization and sprouting are concomitant processes in V. herbacea and can be induced by shoot excision at any phenological phase. 1-FEH and 1-FFT seemed to act in a concerted way to catalyse fructan depolymerization, while 1-SST was inhibited, possibly due to interruption of sucrose supply to rhizophores from the aerial organs.

Fructans↗