Behavioural studies on the ethanol leaf extract of Grewia carpinifolia in Wistar rats.

Background: Grewia carpinifolia is a plant commonly used in the tropics to manage various central nervous system (CNS) disorders. However, despite its widespread use no scientific work has been reported to validate these claims. Objectives: To evaluate the activity of G. carpinifolia as it affects behaviour using animal model. Methods: Twenty five adult Wistar rats were randomly divided into five groups (A-E). Group A served as control (given only distilled water), Groups B,C, D and E were administered with single oral dose of ethanol extract of G. carpinifolia leaf at 100, 200, 400 and 800 mg/kg body weight respectively for twenty eight days consecutively. Subsequently, open field test, negative geotaxis and hanging wire test were performed. Body and brain weights were measured and histological examination of the brain was also performed. Results: At the tested doses, the extract significantly increased the time spent on the hanging wire and decreased locomotor activity at 800 mg/kg. No significant difference was observed in body and brain weights of extract treated groups when compared with the control. No visible histological lesion was also observed. Conclusion: The plant extract may improve muscular strength at tested doses and possess CNS depressant activity at 800 mg/ kg.

Anti-nociceptive and anti-inflammatory potentials of fractions from the extract of Tetracarpidium conophorum leaf in rats and mice

Tetracarpidium conophorum (TC) (Euphorbiaceae) is a perennial woody climbing shrub in low bush forest of some parts of West Africa and used among the natives for relief of ailments accompanying pain and inflammation. In this study, the analgesic and anti-inflammatory effects of the methanolic extract (METC) and fractions (ethyl acetate, F1 and n-hexane, F2) of Tetracarpidium conophorum leaf were evaluated in rat and mice. The analgesic activity was evaluated using acetic acid-induced writhing, formalin-induced paw licking and hot plate test models. Carrageenan-induced paw oedema was used to assess anti-inflammatory activity in rats. The mechanism of action of (TC) was explored by the use of naloxone, a non-selective opioid receptor blocker. The highest analgesic effect was observed in F2 extract at 57.21% inhibition and was further studied on various analgesic and anti-inflammatory models in graded doses. F2 significantly inhibited the late phase of formalin-induced paw licking and prolong hot plate latency at 55±1°C. The n-hexane fraction also significantly inhibited carrageenan-induced paw oedema in rats at 100 and 200mg/kg doses significantly (p< 0.001) and reduced paw licking response by 85.08% compared with control. Naloxone, an opioid receptor antagonist, did not significantly affect the changes observed with n-hexane fraction, thus ruling out the possibility of the involvement of opioid receptors in the analgesic actions of Tetracarpidium conophorum. Phytochemical screening showed that the leaf extracts contain alkaloids, tannins, saponins and cardenolides. The investigations showed that Tetracarpidium conophorum possesses significant anti-nociceptive and anti-inflammatory activities that should be explored.

Determination of antioxidant activity, phenolic contents and antiviral potential of methanol extract of Euphorbia spinidens (Euphorbiaceae)

Purpose: This study was aimed to evaluate the antioxidant activity of the methanol extract of Euphorbia spinidens (Euphorbiaceae) and its effect on Herpes simplex virus type-1 (HSV-1) replication. Methods: The methanol extract of aerial parts of E. spinidens collected from Khorasan State in North- Eastern part of Iran was used in this study. Total phenolic, flavonoid contents and the antioxidant activity were evaluated using Folin-Ciocalteu method, aluminum chloride colorimetric method and β- carotene-linoleate model system, respectively. Both the cytotoxic and antiviral effects of the crude extract on Vero cell line were determined by quantifying the viability of Vero cells using 3-(4,5- dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)2H-tetrazolium (MTS) assay. Results: Total phenolic and flavonoid contents of E.spinidens were 70 ± 1 mg of gallic acid equivalent/g of dry extract (mg GAE/g extract) and 49.66 ± 1.00 mg rutin equivalent/g of dry extract (mg RTN/g extract), respectively. Antioxidant activity was 44 ± 1 % compared with the standard, buthylated hydroxytuloene (BHT). The 50 % cytotoxic concentration (CC50) of the extract on Vero cells was 5.072 ± 0.063 mg/ml and its antiviral concentration of 50 % effectiveness (EC50) value was 0.34 ± 0.003 mg/ml. Conclusion: The findings of this study show that the methanol extract of E. spinidens has high content of phenolic and flavonoid compounds with good antioxidant activity. Furthermore, this extract has significant antiviral effect on HSV-1 probably due to the inhibition of viral replication.

Anti-anxiety effect of methanol extract of Pericarpium zanthoxyli using a strychnine-sensitive glycine receptor model

Purpose: To determine if the methanol extract of Pericarpium zanthoxyli exerts anti-anxiety effects and also to explore any probable anti-anxiety mechanism in vivo. Methods: The staircase test, elevated plus maze test, rota-rod treadmill test and convulsions induced by strychnine and picrotoxin on mice were tested to identify potential mechanism of anti-anxiety activity of the plant extract. Results: The plant extract (10 mg/kg, p.o.) significantly reduced rearing numbers in the staircase test while it increased the time spent in the open arms as well as the number of entries to the open arms in the elevated plus maze test, suggesting that it has significant anti-anxiety activity. Furthermore, the extract inhibited strychnine-induced convulsion. However, it had little effect on picrotoxin-induced convulsion, suggesting that its anti-anxiety activity may be linked to strychnine-sensitive glycine receptor and not GABA receptor. Conclusion: These results suggest that the Pericarpium zanthoxyli extract may be beneficial for the control of anxiety.

Chromatographic-mass spectrometric analysis of ethanol extract of Maesa perlaria var formosana

Purpose: This study analyzes the chemical composition of ethanol root extracts of Maesa perlaria var formosana by gas chromatography-mass spectrometry (GC-MS). Methods: The dried root of Maesa perlaria var formosana was extracted with 95 % ethanol for composition analysis under the following optimum GC-MS conditions: 250 °C inlet temperature, 250 °C MSD detector temperature, and GC oven temperature programmed as follows: initial temperature held at 70 °C for 15 min, then increased at a rate of 2.5 °C/min and held at 170 °C for 15 min; then raised at a rate of 2 °C/min and kept at 180 °C for 20 min; then raised at 2 °C/min and kept at 250 °C for 20 min. Finally, it was raised at 3 °C/min and kept at 280 °C for 15 min. Results: A total of 59 chemical compounds were identified, representing 88.82 % of the composition of the ethanol extracts. The three major components, include 2,4-di-tert-butylphenol (16.76 %), stigmasterol (15.86 %) and campesterol (7.33 %). Conclusion: The results show that a total of 59 components were identified in the ethanol extract of Maesa perlaria var. formosana. The major component, 2,4-Di-tert-butylphenol, exhibits various biological activities.

Antipyretic, analgesic and anti-inflammatory activities of methanol extract of root bark of Acacia jacquemontii Benth (Fabaceae) in experimental animals

Purpose: To investigate the ethnomedicinal claims regarding the use of Acacia jacquemontii Benth. (Fabaceae) in fever, pain and inflammation. Methods: The methanol root bark extract (AJRBM) of the plant was used in the studies. Preliminary phytochemical screening of the extract was carried out according to established methods. Analgesic, anti-inflammatory and antipyretic activities were evaluated using acetic acid-induced writhing, carrageennan-induced rat paw edema and Brewer’s yeast-induced pyrexia models, respectively. The extract was administered at doses of 50 and 100 mg/kg. Aspirin (300 mg/kg, p.o.) was used as a reference drug in all models. Normal saline (10 mL/kg p.o.) was used as negative control. Results: Phytochemical screening results indicate the presence of cardioactive glycosides, tannins, flavonoids and saponins. In the acetic acid-induced writhing model, the methanol extract exhibited significant (p < 0.05) analgesic effect with 58.98 % reduction in writhing response at a dose of 100 mg/kg, compared with untreated control group. The extract significantly (p < 0.05) reduced carrageenan-induced edema at doses of 50 and 100 mg/kg to 36.84 and 47.36 %, respectively, after 1 h of extract administration. The extract exhibited predominantly dose-dependent antipyretic effect in Brewer’s yeast-induced pyrexia model. Maximum reduction in body temperature to 37.07 and 38.29 ºC at doses of 50 and 100 mg/kg, respectively, was observed, compared with untreated group (38.90 ºC) after 1 h, but this was not significant (p < 0.05). Conclusion: The plant extract exerts inhibitory effect on peripheral pain stimuli, edema and dosedependent anti-pyrexia, and thus justifies the ethnomedicinal use of Acacia jacquemontii Benth. in the management of pain, fever and inflammation.

Immunomodulatory activity of methanol extract of Adansonia digitata L

Purpose: To evaluate the immune-modulatory activities of various plant parts Adansonia digitata L. using delayed-type hypersensitivity rat model. Methods: Defatted leaf, root bark and fruit pulp of A. digitata were extracted with methanol. Immunomodulatory activity of the methanol extracts (250 and 500 mg/kg) were evaluated in sheep RBC (SRBC)-induced delayed-type hypersensitivity model, cell mediated immune re-sponse and phagocytic activity using carbon clearance test. Results: The extracts exhibited significant increase in delayed-type hypersensitivity reaction, indicating the ability of the extracts to stimulate T-cells. It also increased SRBC induced anti-body titer in immunesuppressed rats, and produced significant increase in phagocytic index by rapid removal of carbon particles from the blood stream. Conclusion: These results indicate that methanol extracts of the leaf, root bark and fruit pulp of A. digitata hold promise as immunemodulatory agents.

Evaluation of a root extract gel from Urtica dioica (Urticaceae) as analgesic and anti-inflammatory therapy in rheumatoid arthritis in mice

Purpose: To develop and characterize an herbal gel prepared from methanol root extract of Urtica dioica (Urticaceae) (Stinging nettle) for the treatment of arthritis in mice. Methods: A methanol root extract from Urtica dioica was prepared, and a gel was then prepared using Carbopol 934. The prepared gel was subjected to various physical tests (color, appearance, pH, texture, viscosity) and in vivo evaluation, including primary skin irritation, analgesic, and anti-inflammatory tests, in arthritic mice and compared with 2 % indomethacin gel, which was used as standard. Results: The prepared herbal gel was of light gray color with a smooth texture. It showed a pH of 7.1 and a viscosity of 21.2 cps. The gel exhibited pseudoplastic rheology, as evidenced by shear thinning with increased shear rate. It was non-irritating to the skin in primary skin irritation test in mice and showed 55.05 % inhibition of paw edema in a carrageenan-induced hind rat paw edema model, comparable to that of the standard gel (53.93 %), after 24 h. The gel showed 58.21 % analgesia, versus 61.19 % analgesia for the indomethacin gel standard in writhing test. Conclusion: The topical gel from methanol root extract of U. dioica may be an efficacious and safe alternative to non-steroidal anti-inflammatory drugs in the treatment of rheumatoid arthritis but this requires further investigations to ascertain its safety and clinical efficacy.

Effect of Arisaema erubescens (Wall) Schott rhizome extract on rheumatoid arthritis

Purpose: To investigate the anti-arthritic activity of the water extract of Rhizoma Arisaematis (WERA) using a collagen II -induced arthritis (CIA) rat model. Methods: CIA was induced in male Sprague-Dawley rats by intradermal injection of bovine collagen II in Complete Freund’s Adjuvant. The rats were treated with daily oral doses of WERA (100, 200, and 400 mg/kg) for 21 consecutive days. Methotrexate (MTX, 3 mg/kg), used as a positive control, was administered orally 2 times/week for 3 weeks. The severity of arthritis was evaluated using indices of paw swelling, arthritic score, body weight, thymus index, and spleen index. In addition, the serum levels of IL-1β, IL-6, IL-10, and TNF-α were measured. Results: All doses of WERA significantly inhibited paw edema (p < 0.01), decreased arthritis scores (p < 0.01) and spleen index (p < 0.05), and alleviated the weight loss associated with CIA in rats. Furthermore, TNF-α, IL-1β, and IL-6 serum levels were significantly decreased (p < 0.05) by all doses of WERA. By contrast, IL-10 serum levels were markedly increased (p < 0.05). Conclusion: WERA exerts therapeutic effects in CIA in rats by decreasing the serum levels of TNF-α, IL-1β, IL-6 and IL-10, suggesting WERA may be an effective candidate drug for treating human rheumatoid arthritis.

Apoptotic properties of Citrus sudachi Hort, ex Shirai (Rutaceae) extract on humanA549 and HepG2 cancer cells

Purpose: To investigate whether Citrus sudachi harvested at two stages of maturity can induce toxicity in a cell-specific manner and to determine the possible mechanisms of Citrus sudachi-induced cytotoxic responses in two types of cancer cells (human lung adenocarcinoma A549 and hepatocellular carcinoma HepG2 cells) and two normal cell lines (lung 16HBE140- and liver CHANG cells). Methods: 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and annexin V/propidium iodidle assay were used to test the antiproliferative activity and apoptosis of methanol extract of Citrus sudachi, respectively. Griess reaction and reverse transcriptase-polymerase chain reaction (RT-PCR) were carried out to evaluate nitric oxide (NO•) production and the mRNA levels of inhibitors of apoptosis (IAP). Results: Citrus sudachi exerted cytotoxicity in a time-dependent manner in cancer cells which increased with increase in maturity but did not affect normal cells. Citrus sudachi was found to induce accumulation of cells in the sub-G1 cell cycle phase, fragmentation of DNA and cell death with characteristics of apoptosis, in both types of cancer cells. Moreover, Citrus sudachi upregulated cellular NO• produced by activation of nitric oxide synthase (NOS), while it suppressed the levels of IAP mRNA in both types of cancer cells. Conclusion: The results obtained suggest that Citrus sudachi induces apoptosis in A549 and HepG2 cells, which may be mediated by NO•. There is need for further studies on the role of Citrus sudachi in cancer treatment.

Anti-cancer and antioxidant properties of phenolics isolated from Toona sinensis A Juss acetone leaf extract

Purpose: To investigate the antioxidant and anticancer activities of phenolics from the leaf extract of Toona sinensis (TS). Methods: Acetone leaf extract of TS was screened for total phenolic and flavanoid contents, and the flanonoids were subjected to high performance liquid chromatographic (HPLC) analysis. Antioxidant properties were assessed via oxygen radical absorbance capacity (ORAC), peroxyl radical scavenging capacity (PSC) and cellular antioxidant activity (CAA), while anti-proliferative activity ins HepG2 cell line was assessed using methylene blue assay. Results: The extract contained 36.02 ± 0.24 mg of gallic acid equiv/g dry weight (DW) and 20.24 ± 1.73 mg of catechin equiv/g DW of total phenolic and total flavonoid, respectively. The levels of rutin and quercitrin were 0.51 and 19.55 mg/g, respectively. Epicatechin, gallic acid, quercitin, isoquercetin were not detected. The extract showed significant antioxidant potential and high anti-proliferation capacity with low cytotoxicity against HepG2 cell in vitro. The underlying mechanism of anti-proliferative effect was induction of apoptosis. Conclusion: TS leaf extract possesses significant in vitro antioxidant properties and anti-proliferative effect against HepG2 cells, which make it a potential anticancer drug source.

Effect of Astragalus membranaceus (Fisch) Bunge extract on streptozocin-induced diabetic in rats

Purpose: To investigate the effect of Astragalus membranaceus (Fisch.) Bunge. extract (AMBE) on streptozotocin-induced diabetic rats. Methods: The aqueous extract of AMB was obtained by steeping the dried Astragalus membranaceus (Fisch.) Bunge. in water at 60 oC three times, each for 1 h, before first drying in an oven at 100 oC and then freeze-drying the last extract thus obtained. Diabete model rats was induced by a single intraperitoneal injection of a freshly prepared solution of streptozotocin (50 mg/kg). The rats were randomly divided into 6 groups of ten rats each: negative control group, normal control group, reference group (glibenclamide1 mg/kgbody weight) as well as AMB extract groups, namely, 40, 80 and 160 mg/kg body weight. Antihyperglycemic effect was measured by blood glucose and plasma insulin levels. Oxidative stress was evaluated in liver and kidney by antioxidant markers, viz, lipidperoxidation (LPO), superoxide dismutase (SOD), reduced glutathione (GSH), glutathione peroxidase (GPx) and catalase (CAT), while blood serum levels of creatinine and urea were also determined in both diabetic control and treated rats. Results: Compared with diabetic rats, oral administration of AMBE at a concentration of 160 mg/kg daily for 30 days showed a significant decrease in fasting blood glucose (109.438 ± 3.52, p < 0.05) and increased insulin level (13.96 ± 0.74, p < 0.05). Furthermore, it significantly reduced biochemical parameters (serum creatinine, 0.86 ± 0.29, p < 0.05) and serum urea (45.14 ± 1.79, p < 0.05). The treatment also resulted in significant increase in GSH (49.21 ± 2.59, p < 0.05), GPx (11.96 ± 1.16, p < 0.05), SOD (14.13 ± 0.49, p < 0.05), CAT (83.25 ± 3.14, p < 0.05) level in the liver and kidney of diabetic rats. Conclusion: The results suggest that AMBE may effectively normalize impaired antioxidant status in streptozotocin-induced diabetes in a dose-dependent manner. AMBE has a protective effect against lipid peroxidation by scavenging free radicals and is thus capable of reducing the risk of diabetic complications.

Effects of aqueous extract from Asparagus officinalis L. roots on hypothalamic-pituitary-gonadal axis hormone levels and the number of ovarian follicles in adult rats

Background: Asparagus is a plant with high nutritional, pharmaceutical, and industrial values. Objective: The present study aimed to evaluate the effect of aqueous extract of asparagus roots on the hypothalamic-pituitary-gonadal axis hormones and oogenesis in female rats. Materials and Methods: In this experimental study, 40 adult female Wistar rats were divided into five groups, which consist 8 rats. Groups included control, sham and three experimental groups receiving different doses (100, 200, 400 mg/kg/bw) of aqueous extract of asparagus roots. All dosages were administered orally for 28 days. Blood samples were taken from rats to evaluate serum levels of Gonadotropin releasing hormone (GnRH), follicular stimulating hormone (FSH), Luteinal hormone (LH), estrogen, and progesterone hormones. The ovaries were removed, weighted, sectioned, and studied by light microscope. Results: Dose-dependent aqueous extract of asparagus roots significantly increased serum levels of GnRH, FSH, LH, estrogen, and progestin hormones compared to control and sham groups. Increase in number of ovarian follicles and corpus luteum in groups treated with asparagus root extract was also observed (p<0.05). Conclusion: Asparagus roots extract stimulates secretion of hypothalamic- pituitary- gonadal axis hormones. This also positively affects oogenesis in female rats.

Effects of Ageratum Conyzoides on Semen Characteristics and Sperm Morphology in Rats Exposed to Sodium Arsenite

Arsenite is a major environmental toxicant that is well known to cause reproductive injury. The sperm protective potential of Ageratum conyzoides Linn in arsenic-treated rats was carried out in this study taking advantage of the antioxidant constituents and its androgenic activities. Twenty-four male albino rats aged 16 weeks, weighing 225 to 228g were used. They were grouped into 4(A-Da) with each group containing 6 rats. Group A was orally treated with 100mg/kg ethanol leaf extract of Ageratum conyzoides L., daily for 14 days, group B (single oral dose of sodium arsenite 2.5 mg/kg body weight), C (Ageratum conyzoides extract daily for 14 days and sodium arsenite (SA) given on the 14th day) and group D (Propylene glycol as negative control). It was observed that group B had a more lower (p<0.05) percentage motility (26.7±6.67%) when compared across the groups while group A had a significantly higher (p<0.05) mean value (63.3±3.33%). The sperm motility of rats in group D was significantly higher (p<0.05) than groups B and C. This implies that A. conyzoides extract had no adverse effect on the sperm motility of the rats and also ameliorates the adverse effect of arsenite on sperm motility. The mean value obtained for sperm liveability, semen volume and Sperm concentration followed a similar pattern although, the differences were not significant (p>0.05) for semen volume and the Sperm concentration of rats across the groups. The total sperm abnormality obtained across the groups ranges between 10.44 and 14.27% with group B treated with sodium arsenite (SA) having the highest value when compared with groups A and D, although, the differences were not significant (P>0.05). The study concluded that ethanol leaf extract of Ageratum conyzoides has no negative effect on sperm motility, liveability characteristics and morphology and also protected spermatozoa against arsenic reproductive toxicity in wistar strain albino rats..

STANDARDIZATION OF CASSAVA MAHEWU FERMENTATION AND ASSESSMENT OF THE EFFECTS OF IRON SOURCES USED FOR FORTIFICATION

Cassava root is the main staple for 70% of the population in Mozambique, particularly in inaccessible rural areas, but is known to be low in iron. Anaemia is a public health problem in mothers and preschool children in Mozambique and up to 40% of these cases are probably due to dietary iron deficiency. The World Health Organization (WHO) and Food and Agriculture Organization of the United Nations (FAO) recognize the fortification of foodstuff as an effective method to remedy dietary deficiencies of micronutrients, including iron. Cassava mahewu, a non-alcoholic fermented beverage is prepared at subsistence level from cassava roots using indigenous procedures. The aim of the study was to standardize mahewu fermentation and investigate if the type of cassava fermented, or the iron compound used for fortification affected the final product. Roots of sweet and bitter varieties of cassava from four districts (Rapale, Meconta, Alto Molocue and Zavala) in Mozambique, were peeled, dried and pounded to prepare flour. Cassava flour was cooked and fermented under controlled conditions (45°C for 24 h). The fermentation period and temperature were set, based on the findings of a pilot study which showed that an end-point pH of about 4.5 was regularly reached after 24 h at 45°C. Cassava mahewu was fortified with ferrous sulfate (FeSO4.7H2O) or ferrous fumarate (C4H2FeO4) at the beginning (time zero) and at the end of fermentation (24 h). The amount of iron added to the mahewu was based on the average of the approved range of iron used for the fortification of maize meal. The mean pH at the endpoint was 4.5, with 0.29% titratable acidity. The pH and acidity were different to those reported in previous studies on maize mahewu, whereas the solid extract of 9.65% was found to be similar. Lactic acid bacteria (LAB) and yeast growth were not significantly different in mahewu fortified with either of the iron compounds. There was no significant difference between cassava mahewu made from bitter or sweet varieties. A standard method for preparation and iron fortification of cassava mahewu was developed. It is recommended that fortification occurs at the end of fermentation when done at household level.