Effect of Aqueous Extract of Unripe Musa Paradisiaca Linn on Parameters Affecting Reproduction in Rats.

OBJECTIVE: Aqueous extract of unripe Musa paradisiaca fruit is commonly used for the treatment of ulcers in eastern Nigeria. This study aimed to assess the acute and subacute effects of an aqueous extract of unripe fruit on male and female fertility in rats. METHODS: Aqueous extracts obtained by maceration were analyzed for acute and subacute toxicity and for the presence of phytochemical constituents using standard procedures. The extract (100, 500, and 1000 mg/kg) was administered daily to rats of both sexes for 28 d. Blood samples collected on days 0 and 28 were assessed for follicle-stimulating hormone (FSH), luteinizing hormone (LH), catalase (CAT), superoxide dismutase (SOD), and malondialdehyde (MDA). Testes and ovaries were harvested for histopathological analysis. Sperm were also collected to determine the sperm count and motility. RESULTS: Phytochemical screening revealed the presence of saponins, tannins, alkaloids, and resins. After an oral dose of up to 5000 mg/kg, there were no deaths in the acute toxicity test. The extract (500 mg/kg) significantly (P < .05) enhanced sperm count and motility relative to the untreated control; significantly (P < .05) reduced SOD, CAT, and glutathione levels, while significantly (P < .05) elevated LH, FSH, and MDA levels in male and female rats. Histological examination revealed significant structural damage to the ovaries. CONCLUSION: Unripe Musa paradisiaca fruit exhibited an adverse toxicological profile following prolonged administration and caused oxidative stress in rodents.

Newboulasides A and B, two new caffeic acid glycosides from Newbouldia laevis with α-amylase inhibitory activity.

Chemical investigation of the ethanol extract of the leaves of Newbouldia laevis (P. Beauv) led to the isolation of two new caffeic acid glycosides, Newboulasides A (1) and B (2). The structures of these compounds were determined on the basis of extensive spectroscopic methods, including 1D-, 2D-NMR and MS data. The extracts and fractions and the isolated compounds were evaluated for their inhibition of α-amylase enzyme activity. The extract showed inhibition of α-amylase activity with IC50 value of 102.91 µg/mL, while the isolated compounds (1 and 2) exhibited pronounced inhibition with IC50 values of 4.95 and 4.44 µg/mL respectively, comparable to the standard - Acarbose with IC50 value of 4.05 µg/mL. Our findings demonstrated that the inhibition of α-amylase activity may be part of the mechanisms through which N. leavis exhibits antidiabetic effect.

Millettia aboensis ameliorates oxidative stress through synergic interaction of its active compounds.

Background M. aboensis has wide ethnopharmacological applications but very little has been done on the pharmacological basis for these indications. This study evaluated the antioxidant potentials of the leaf extracts of M. aboensis. Methods Total phenolic content of the extract and fractions was carried out using folin-ciocalteu method while in vivo site specific effect determined using carbon tetrachloride (CCl4)-induced liver oxidative damage. Chromatographic separations of the most active fraction led to the isolation of compounds 1 and 2 with their structures elucidated by a combination of 1D and 2D NMR and mass spectrometry. Inhibition of liver microsome lipid peroxidation was used to evaluate the antioxidant activities of these compounds while DPPH test was used to study their interaction. Results Ethyl acetate fraction had the highest phenolic content of 305.2 mgGAE/g with n-hexane fraction having the least (26.1 mgGAE/g). Structural elucidation revealed compound 1 as epicathechin-(2ß→O→7, 4ß→8)-cathechin and compound 2 as epicathechin-(2ß→O→7, 4ß→8)-epicathechin. Compounds 1 & 2 inhibited liver microsome lipid peroxidation with EC50 of 46 and 55 µg/mL respectively. Combination of the compounds produced synergic inhibition of DPPH radical with EC50 of 7 µg/mL against 9 µg/mL produced by ascorbic acid. Conclusion M. aboensis expressed strong antioxidant property which may explain its diverse ethnopharmacological uses.

Newbouldia laevis improved glucose and fat homeostasis in a TYPE-2 diabesity mice model.

ETHNOPHARMACOLOGICAL RELEVANCE: Newbouldia laevis (P. Beauv) has a long-standing ethnomedicinal use in the management of diabetes mellitus. However, no scientific evidence has established its potentials in the management of obesity dependent diabetes. AIM: This study aimed at filling the gap in knowledge about the antidiabetic activity of the leaf extract of N. laevis in a type-2 diabesity mice model. MATERIALS AND METHOD: The ethanol leaf extract was subjected to liquid-liquid partitioning successively with n-hexane, ethyl acetate and butanol to obtain respective fractions soluble in these solvents. The butanol (most active) fraction at 302 and 604 mg/kg was further tested on high-fat diet STZ-NAD induced type-2 diabetic mice for 10 weeks with glibenclamide (10 mg/kg) and pioglitazone (30 mg/kg) as standards. The effect on food intake, body weight, fasting blood glucose, oral glucose tolerance test (OGTT), oral fat tolerance test (OFTT), insulin secretion, insulin resistance and lipid profile were determined prior to treatment, mid-way and at the end of 10 weeks treatment. RESULTS: Higher food consumption was recorded in diabetic (D) animals on high-fat diet (HFD) compared to the normal diet (ND)-fed groups. Treatment of these diabetic mice on HFD with 604 mg/kg of butanol fraction produced significant (p < 0.05) reduction in body weights of these animals from the 2nd week to the 9th week. Ten weeks treatment with butanol fraction achieved a marked decrease in blood glucose and also an increase in fat clearance. Just like pioglitazone, treatment with butanol fraction at both 302 and 604 mg/kg doses produced significant (p < 0.05) decrease in HFD mediated elevation of serum insulin and a non-significant (p > 0.05) increase in STZ-NAD mediated depletion of serum insulin. Butanol fraction at 604 mg/kg also produced reduction in insulin resistance as indicated by significant (p < 0.05) decrease in HOMA-IR value on the 5th and 10th week just like pioglitazone (30 mg/kg). CONCLUSION: N. laevis exhibited wide actions in the regulation of glucose and fat homeostasis making it a potential novel agent for the management of diabetes, obesity and their likely associated complications.

Upregulation of CD4+ T-Lymphocytes by Isomeric Mixture of Quercetin-3-O-Rutinoside and Quercetin-3-O-Robinobioside Isolated from Millettia aboensis.

BACKGROUND: Millettia aboensis (Hook. F.) Baker (Fabaceae) is popular in ethnomedicine for its acclaimed efficacy in a number of disease conditions. This study evaluated the immunomodulatory effect of the leaf extract as a possible mechanism of its ethnomedicinal uses. METHODS: Humoral and cellular immune responses of Balb/c mice to tetanus toxoid and cyclophosphamide, respectively, were used to monitor immunomodulatory activities of the ethanol leaf extract and fractions of M. aboensis at 200, 300 and 400 mg/kg. Active (butanol) fraction of the extract was subjected to chromatographic purifications to isolate the active compound and the structure elucidated by a combination of 1D and 2D NMR and mass spectrometry. Stimulation of specific T-lymphocytes using intracellular cytokine staining technique was used to evaluate immune-enhancing activity of the isolated compound. RESULTS: The extract and fractions evoked increase in both humoral and cellular immunity. At 400 mg/kg of butanol fraction, the normalized mean secondary production of IgG1 and IgG2a antibodies were 9.0 and 7.7, respectively. Serum cytokine production by butanol fraction following secondary challenge with tetanus toxoid showed that IL-12, IL-17A and IFN-γ were expressed by 48.14, 41.37 and 38.22%, respectively. Structural elucidation of the active compound revealed presence of isomeric mixtures of quercetin-3-O-rutinoside and quercetin-3-O-robinobioside (Compound 1a/b). Compound 1a/b exhibited in vitro upregulation of specific CD4+ T-lymphocytes that were largely IFNγ releasing with up to 43.7% stimulation at 6.25 µg/mL compared to the baseline effect in DMSO vehicle control group. CONCLUSION: M. aboensis expressed strong immune-enhancing properties, which may explain its ethnopharmacological use in disease management.

Anti-plasmodial and anti-inflammatory activities of cyclotide-rich extract and fraction of Oldenlandia affinis (R. & S.) D.C. (Rubiaceae).

BACKGROUND: Oldenlandia affinis, commonly called 'kalata-kalata', a versatile plant used locally to treat malaria fever in some parts of sub-Saharan Africa was investigated for anti-plasmodial and anti-inflammatory activities. OBJECTIVE: The study was designed to evaluate the antiplasmodial as well as anti-inflammatory activities of whole extract and cyclotide-rich fraction of Oldenlandia affinis. METHOD: The dichloromethane-methanol extract (ODE) of the plant, O. affinis was investigated for suppressive and curative antiplasmodial activities against Plasmodium berghei in mice. ODE and the cyclotide-rich fraction (CRF) was investigated for chronic and acute anti-inflammatory activities in rat models of inflammation. Inhibition of pro-inflammatory mediators was studied in RAW264.7 macrophages. RESULTS: ODE exhibited significant (p<0.05) reduction in mean parasitaemia in both the suppressive and curative models of Plasmodium berghei infection in mice.Administration of ODE(100, 200, or 400 mg/kg) and CRF (100, 200, or 400 mg/kg) produced significant inhibition of rodent models of acute and chronic inflammation . This observation is supported by the significant (P<0.05) inhibition of pro-inflammatory mediators, inducible nitric oxide (iNO) and tumour necrosis factor-alpha (TNF-α), and the reactive radical scavenging activities in RAW264.7 macrophages. CONCLUSION: These findings could explain, at least in part, the successes reported in the use of the herb, Oldenlandia affinis in the traditional treatment of malaria fever.

Landolphia owariensis leaf extracts reduce parasitemia in Plasmodium berghei-infected mice.

Context Landolphia owariensis P. Beauv. (Apocyanaceae) leaf is used in southeast Nigeria to treat malaria. Objective This study evaluated the antiplasmodial activity of L. owariensis leaf extract and fractions, also the phytoconstituents were standardized and analyzed. Methods The effects of daily, oral administrations of 200, 400 and 800 mg/kg of L. owariensis leaf extract (LOE), its hexane (LOHF), ethyl acetate (LOEF) and methanol (LOMF) fractions on early, established and residual infections in Plasmodium berghei-infected albino mice were evaluated in vivo. The extract and fractions were subjected to phytochemical analysis and HPLC fingerprinting, and the acute toxicity of LOE was evaluated. Results The extract and fractions elicited 29-86, 18-95 and 75-96% significant (p < 0.001) suppression of parasitemia in early, established and residual infections, respectively. The ED50 values for suppressive activity of LOE, LOHF, LOEF and LOMF were 266.56, 514.93, 392.95 and 165.70 mg/kg, respectively. The post-day 30-survival index was 16.7-50, 16.7, 16.7-66.7 and 50-83.3% for LOE, LOHF, LOEF, and LOMF, respectively. Extract-treated mice significantly (p < 0.001) gained weight and had reduced mortality compared with negative control (untreated) mice. An oral LD50 value >5000 mg/kg in mice was established for LOE. The LOMF showed the greatest antiplasmodial activity in all the models, suggesting that the antimalarial activity of the plant may be attributed to alkaloids, flavonoids, saponins and tannins present in the fraction. Conclusion Results demonstrate the antiplasmodial activity of L. owariensis leaf, and provide a pharmacological rationale for its ethnomedicinal use as an antimalarial agent.

Extracts of Leptadenia hastata Leaf, a Famine Food and Traditional Remedy for Furuncles, Suppress Inflammation in Murine Models.

The effects of the methanol extract (LHE), hexane (LHHF), ethylacetate (LHEF) and methanol (LHMF) fractions of leaf of Leptadenia hastata on acute and chronic inflammation were studied. Furthermore, the effects of LHE on acetic acid induced increase in vascular permeability, carrageenan induced leucocyte migration and membrane stability were evaluated. The LHE and fractions were also subjected to phytochemical analysis. The LHE, LHEF and LHMF significantly (p < 0.05) suppressed topical ear edema, systemic paw edema, global edematous response to formaldehyde arthritis and granuloma tissue growth. The LHE suppressed acetic acid induced vascular permeability and carrageenan-induced leucocyte migration, and also stabilized the erythrocyte membrane. An acute toxicity test in mice established an oral LD50 > 5 g/kg for LHE. The LHEF elicited the greatest inhibition, suggesting that the observed anti-inflammatory effects may be attributable to the flavonoids abundant in the fraction. These findings demonstrate that the effectiveness of L. hastata leaf in the treatment of furuncles may largely derive from anti-inflammatory activities mediated through inhibition of both increase in vascular permeability and leucocyte migration, and stabilization of cell membranes.

Wound-healing Activity of the Aqueous Leaf Extract and Fractions of Ficus exasperata (Moraceae) and its Safety Evaluation on Albino Rats.

Ficus exasperata have been reported to have wide applications in the treatment of many human diseases. However, its traditional use in the treatment of wounds has not been validated by any scientific study. Also, its safety in the management of chronic disease conditions requires attention. We evaluated the wound-healing activity of the aqueous extract and fractions of F. exasperata, as well as its safety after subchronic oral administration. Similar percentage of wound contraction was observed with 5% w/w extract ointment application and administration of cicatrin powder (standard) on the 4(th) day, while better contraction than the standard was recorded with higher concentrations of the extract ointment. Of all the fractions tested, significant (P < 0.05) contraction was only noticed in chloroform fraction, though lower than that of the aqueous extract. The extract also showed concentration-dependent inhibition of all the tested microbial isolates. Extract administered up to 5000 mg/kg (single dose administration) did not cause any mortality after 24 h. Mortality was, however, recorded at 4000 mg/kg within the first 20 days of subchronic administration of the extract. Significant (P < 0.05) increases in alanine aminotransaminase (ALT), aspartate aminotransaminase (AST), and in particular, alkaline phosphatase (ALP) were observed at different doses and time periods. Pathological and histological changes were noticed in the liver and kidney on the 91(st) day of the study with 4000 mg/kg of the extract. Except for the significant (P < 0.05) reduction in WBC on the 91(st) day, no other significant (P < 0.05) changes were observed in other hematological parameters. The aqueous extract demonstrated better wound-healing activity than its fractions; however, the extract may not be safe at higher doses for subchronic oral administration, as may be the case in the management of chronic disease conditions.

Antimicrobial Effects of a Lipophilic Fraction and Kaurenoic Acid Isolated from the Root Bark Extracts of Annona senegalensis.

Root bark preparation of Annona senegalensis Pers. (Annonaceae) is used in Nigerian ethnomedicine for treatment of infectious diseases. Extraction of the A. senegalensis powdered root bark with methanol-methylene chloride (1 : 1) mixture yielded the methanol-methylene extract (MME) which was fractionated to obtain the ethyl acetate fraction (EF). The EF on further fractionation gave two active subfractions, F1 and F2. The F1 yielded a lipophilic oily liquid while F2 on purification, precipitated white crystalline compound, AS2. F1 was analyzed using GC-MS, while AS2 was characterized by proton NMR and X-ray crystallography. Antibacterial and antifungal studies were performed using agar-well-diffusion method with 0.5 McFarland standard and MICs calculated. GC-MS gave 6 major constituents: kaur-16-en-19-oic acid; 1-dodecanol; 1-naphthalenemethanol; 6,6-dimethyl-bicyclo[3.1.1]hept-2-ene-2-ethanol; 3,3-dimethyl-2-(3-methylbuta-1,3-dienyl)cyclohexane-1-methanol; 3-hydroxyandrostan-17-carboxylic acid. AS2 was found to be kaur-16-en-19-oic acid. The MICs of EF, F1, and AS2 against B. subtilis were 180, 60, and 30 µg/mL, respectively. AS2 exhibited activity against S. aureus with an MIC of 150 µg/mL, while F1 was active against P. aeruginosa with an MIC of 40 µg/mL. However, the extracts and AS2 exhibited no effects against Candida albicans and Aspergillus niger. Therefore, kaurenoic acid and the lipophilic fraction from A. senegalensis root bark exhibited potent antibacterial activity.

Studies on the antiulcer and gastrointestinal effects of stem bark extract of Bridelia ferruginea.

The antiulcer and gastrointestinal effects of methanol stem bark extract (BFME) of Bridelia ferruginea Benth. (Euphorbiaceae) and its solvent fractions-dichloromethane (DCMF) and methanol (MF)-were studied using indomethacin- and ethanol-induced ulcers in rats, small intestinal transit of charcoal meal in mice, and the effects on acetylcholine-induced contractions of the isolated guinea pig ileum. The extract and fractions significantly (P<0.05) protected the rats against ethanol and indomethacin-induced ulcers and inhibited small intestinal propulsion in the order of magnitude: DCMF>MF>BFME. On the guinea pig ileum, MF (0.05 - 6.40 mg/ml) elicited no inhibition, DCMF (5 - 40 µg/ml) antagonized acetylcholine-induced contractions of the guinea pig ileum with IC50 of 10.47µg/ml, while BFME (0.05 - 12.80 mg/ml) contracted the guinea pig ileum with EC50 of 1 mg/ml. Oral LD50 of BFME in mice was estimated to be 2,154 mg/kg. Phytochemistry tests revealed the presence of tannins, saponins, steroids, terpenoids, flavonoids and resins in BFME, MF tested positive for tannins, saponins, steroids, terpenoids, and flavonoids, while DCMF gave positive reactions for flavonoids, steroids, terpenoids and resins. These findings suggest that constituents of the stem bark of B. ferruginea possess antiulcer properties. In addition, some non-polar constituents possess spasmolytic activity while spasmogenic activity is likely associated with some polar constituents.

Hypotensive activity of the ethanol extract of Pavetta crassipes leaves.

Pavetta crassipes leaf is routinely used locally in Nigeria for the management of respiratory disorders and hypertension. The hypotensive and other cardiovascular effects of Pavetta crassipes were investigated in cats and rats. The effects of the extract on rat and cat blood pressures, isolated rat atria, rat portal vein, isolated rat aorta and rat vas deferens were studied. Specific receptor antagonists (atropine, mepyramine, phentolamine, propranolol) were used to elucidate the underlying mechanism(s) involved in the cardiovascular changes induced by P. crassipes. The results revealed that the ethanolic extract of Pavetta crassipes lowered the blood pressures of cats and rats in a dose dependent manner. The extract also caused a concentration-dependent decrease in the force of contraction of the isolated rat atria and rat portal vein. The decreases in blood pressure values were attenuated in the presence of a beta-adrenoceptor antagonist, propranolol. The extract also attenuated isoprenaline-induced contraction of the rat atria. However, the extract did not affect contractions evoked by KCl, norepinephrine and 5-HT on the rat aorta. Pavetta crassipes contains biologically active substances that may be useful in the management of hypertension.

Interaction between chloroquine sulphate and aqueous extract of Azadirachta indica A. Juss (Meliaceae) in rabbits.

This study was carried out to investigate the effect of concurrent oral administration of aqueous leaf extract of Azadirachta indica (Meliaceae) on the pharmacokinetic properties of chloroquine sulphate in experimental rabbits. The results indicated that concurrent administration of both agents resulted in a significant decrease in serum concentration, slower absorption and elimination as well as longer half-life of chloroquine sulphate. The highest relative decrease of 78.0% was recorded 4 hours after concurrent administration, while the smallest decrease (64.6%) occurred 24 hours after concurrent administration. Significant reductions were also noted in some pharmacokinetic parameters of chloroquine and included the area under the curve (71.9%), maximum serum concentration (69.8%), absorption rate constant (37.3%), elimination rate constant (53.9%), clearance rate (76.5%) and volume of distribution (47.2%). However, there was a pronounced increase in the half-life of the drug (125.7%).