Ganoderma lucidum ethanol extract promotes weight loss and improves depressive-like behaviors in male and female Swiss mice.

Metabolic and mood-related disturbances can increase the risks of developing adverse mental health problems. The medicinal mushroom, Ganoderma lucidum, is utilized in indigenous medicine to improve quality of life, promote health, and boost vitality. This study investigated the effects of Ganoderma lucidum ethanol extract (EEGL) on feeding behavioral parameters, depressive-like symptoms, and motor activity in Swiss mice. We hypothesized that EEGL would have beneficial effect on metabolic and behavioral outcomes in a dose-related manner. The mushroom was identified and authenticated via techniques of molecular biology. Forty Swiss mice (n = 10/group) of either sex were given distilled water (10 mL/kg) and graded doses of EEGL (100, 200, and 400 mg/kg) orally for 30 days, during which feed and water intake, body weight, neurobehavioral, and safety data were documented. The animals experienced a significant decrease in body weight gain and feed intake while water intake increased in a dose-dependent manner. Furthermore, EEGL significantly diminished immobility time in forced swim test (FST) and tail suspension test (TST). At the 100 and 200 mg/kg, EEGL did not cause significant alteration in motor activity in the open field test (OFT). Meanwhile, an increase in motor activity in male mice without remarkable difference in female mice was observed at the highest dose (400 mg/kg). Eighty percent of mice treated with 400 mg/kg survived till day 30. These findings suggest that EEGL at 100 and 200 mg/kg reduces the amount of weight gained and elicits antidepressant-like effects. Thus, EEGL might be useful for the management of obesity and depressive-like symptoms.

Myrianthus arboreus P. Beauv improves insulin sensitivity in high fat diet-induced obese mice by reducing inflammatory pathways activation.

ETHNOPHARMACOLOGICAL RELEVANCE: Metabolic syndrome is currently recognized as the major cause of morbidity, with dramatic complications on life expectancy and health status. Myrianthus arboreus is a medicinal plant traditionally used in local communities as a safe remedy in treating diabetes and other metabolic diseases. AIM OF THE STUDY: This study aimed to investigate the impact of a methanol extract of Myrianthus arboreus leaf (MAL) in a mice model of metabolic syndrome induced by a high-fat diet (HFD) intake. MATERIALS AND METHODS: Male C57BL/6J mice were assigned to the following groups: control, obese control, and obese treated with MAL extract (10, 25, and 50 mg/kg) for 6 weeks. Control mice received a standard chow diet, while all obese mice were fed with HFD. Animal weight and food consumption were periodically measured. At the end of the treatment, fasting blood glucose and metabolic plasma analysis (insulin level, triglycerides, and total cholesterol (TC)) were performed. The HFD-induced inflammatory status and the expression of several obesity-related markers were evaluated in liver and fat using qPCR and Western blot analysis. In addition, the phytochemical composition of MAL was identified by GC-MS and HPLC-MS. RESULTS: MAL administration significantly reduced body weight gain, basal glycemia, and insulin resistance, and improved plasma lipid profile compared with HFD-fed mice. Similarly, this extract improved the HFD-associated inflammatory status in mice by gene expression modulation of different inflammatory markers involved in this experimentally induced metabolic condition. CONCLUSION: These results demonstrate the novel applicability of MAL, thus suggesting it as a promising therapeutic approach for the management of metabolic disorders.

Beneficial effect of Bidens pilosa L. (Asteraceae) in a rat model of colitis.

Background Bidens pilosa (BP) possessed anti-inflammatory, antioxidant, and immunomodulatory activities. Its beneficial effects on intestinal inflammation and oxidative stress in 2,4,6 trinitrobenzene sulfonic acid (TNBS) induced colitis in Wistar rats was evaluated. Methods Thirty female Wistar rats weighing 180-200 g were distributed into six groups (n = 5): non-colitic, untreated colitic and colitic rats treated graded doses of methanol extract of BP (50-400 mg/kg). Colitis was induced in rats by intracolonic instillation of 0.2 mL of 40 mg/mL TNBS. BP was administered two days pre-colitis induction and treatments continued until seven days post-colitis induction. A day after the last treatment, rats were euthanized, colon removed aseptically and response to treatment assessed. Phytochemical composition of BP was determined using the GC-MS. Results BP significantly reduced macroscopic colonic damage score, weight/length ratio, colonic lipid peroxidation level, leukocytes infiltration, and TNF-α level in comparison to untreated colitic rats (p ≤ 0.008). Similarly, treatment with 200 and 400 mg/kg BP prevented depletion of colonic glutathione level than other treatment groups (p ≤ 0.0002). Histological findings revealed that treatment with 400 mg/kg BP significantly preserved the mucosal epithelial layer. It also prevented ulceration and sloughing of the mucosal layers and reduced infiltration of inflammatory cells compared to other treatment groups. Among the 16 compounds identified were oleic acid (6.2%) and n-hexadecanoic acid (2.0%) with antioxidant anti-inflammatory activities. Conclusions The beneficial effects of BP in rat colitis might be related to the reduction of leucocytes infiltration, inhibition of oxidative stress and pro-inflammatory cytokines.

Ethanol Extract of Blighia sapida Stem Bark Show Remarkable Prophylactic Activity in Experimental Plasmodium berghei-Infected Mice.

This work explores the antiplasmodial potential of ethanol extract of Blighia sapida (Lin. Sapindaceae) in chloroquine (CQ)-resistant Plasmodium berghei (ANKA strain)-infected mice. Chloroquine-resistant (ANKA) strain of P berghei was inoculated intraperitoneally into Swiss albino mice. Mice were treated orally for 4 consecutive days, before and after inoculation (prophylactic, suppressive, and curative models) with graded doses of the plant extracts with Artemether-Lumefantrine (Coartem) as control. Prophylactically, the extract showed a remarkable activity in the chemosuppression of P berghei parasites (P < .01) ranging from 57% to 36.5% at doses of 200 to 800 mg/kg, respectively, whereas Coartem (10 mg/kg) produced 62.1% chemosuppression. No significant chemosuppression was observed in the curative and suppressive models. The plant extract appeared to be safe at the highest dose tested (5000 mg/kg) for acute toxicity, with no adverse effect on the different organs. The plant extract possesses prophylactic antimalarial activity, which supports its use in the prevention of malaria.

Antiinflammatory and immunomodulatory activity of an ethanolic extract from the stem bark of Terminalia catappa L. (Combretaceae): In vitro and in vivo evidences.

ETHNOPHARMOCOLOGICAL RELEVANCE: Terminalia catappa Linn (Combretaceae) is a medicinal plant with anti-inflammatory, anti-diarrhoeal and antioxidant properties, frequently found in tropical regions. Considering its characteristics, it could be useful for the treatment of inflammatory bowel disease, which is associated with inflammation, oxidative stress and an immune dysfunction. Thus this study evaluates the immunomodulatory properties and the intestinal anti-inflammatory effect of an ethanolic extract of the stem bark of T. catappa (ETCB) both in vitro (in RAW 264.7 macrophages) and in vivo, in the trinitrobenzenesulfonic acid (TNBS) model of rat colitis. MATERIALS AND METHODS: The phenolic compounds in ETCB were identified and quantified using HPLC-DAD-qTOF-MS. The immunomodulatory activity ETCB was tested in vitro by determining the macrophage production of IL-1ß and nitrites. In vivo studies were performed in the TNBS model of rat colitis. ETCB was given (25, 50 and 100mg/kg/day) orally for two days prior to colitis induction and thereafter for 7 days. Response to treatment was assessed by scoring the gross appearance of the colon, and determining myeloperoxidase activity, gene expression of pro-inflammatory cytokines like TNF-α, IL-23 and IL-6, chemokines, inducible nitric oxide synthase and proteins crucial in the maintenance of the intestinal mucosal barrier integrity like mucins (MUC-2, MUC-3) and villin. RESULTS: ETCB was able to inhibit IL-1ß and nitrite production in vitro in RAW 264.7 macrophages. Moreover, treatment of TNBS colitic rats with ETCB resulted in a decreased colonic damage score and weight/length ratio. It also reduced the colonic neutrophil infiltration indicated by a lower myeloperoxidase activity and prevented the depletion of colonic glutathione levels in colitic rats. In addition, treatment with ETCB down-regulated the gene expression of pro-inflammatory mediators (TNF-α, IL-23, IL-6 and CINC-1) and iNOS in colitic rats. Moreover, the gene expression of mucosal barrier proteins like MUC-2, MUC-3 and villin were up-regulated in colitic rats treated with ETCB. The dose of ETCB that produced the most significant beneficial effect was 100mg/kg. Regarding the chemical composition of ETCB, 31 phenolic compounds were identified, including ellagic acid, catalagin and gallic acid. CONCLUSION: The beneficial effect of ETCB in the TNBS induced colitis in rats could be related to its antioxidant, immunomodulatory and anti-inflammatory activities, which could be attributed to the phenolic compounds identified.

The effect of lopinavir/ritonavir on the antimalarial activity of artemether or artemether/lumefantrine in a mouse model of Plasmodium berghei.

The possibility of drug-drug interactions occurring during the treatment of malaria infection in human immunodeficient virus (HIV) patients receiving antiretroviral drugs is very high and limited data are available. This study reports the effect of lopinavir/ritonavir (LR) an antiretroviral drug on the antimalarial activity of standard dose of artemether/lumefantrine (AL) or artemether (AM) in a mouse model of Plasmodium berghei. The 50% effective dose (ED50) of AM alone (0.80 ± 0.15 and 2.18 ± 0.75 mg/kg) or in combination with LR (0.88 ± 0.40 and 3.53 ± 1.09 mg/kg) on days 4 and 5 post-infection was similar. In addition, treatment with a standard dose of AL alone or in combination with LR resulted in complete suppression of parasite growth. However, co-administration of LR with AL appears to be toxic resulting in lower survival of experimental animals in comparison to those treated with standard dose of AL alone.

Interaction between rifampicin, amodiaquine and artemether in mice infected with chloroquine resistant Plasmodium berghei.

BACKGROUND: Artemisinin-based combination therapy (ACT) remains the most effective chemotherapeutic strategy in the management of malaria. However, reports of reduced susceptibility of Plasmodium falciparum to the ACT justify the need for continued search for alternative anti-malarial drugs. The use of antibiotics with anti-malarial properties represents a potentially valuable chemotherapeutic option for the management of drug resistant infections. Thus, the intrinsic anti-malarial activity of the combination of clinical doses of rifampicin with amodiaquine and artemether was evaluated in an animal model using Plasmodium berghei. METHODS: A modification of the suppressive tests in vivo was employed. The anti-malarial activity of standard doses of amodiaquine (AQ) with or without artemether (ART) and combined with varying doses of rifampicin (RIF 15 mg/kg or RIF 30 mg/kg body weight) was evaluated in 40 mice sub-divided into eight groups and inoculated intraperitoneally with 1 × 10(7) red blood cells infected with chloroquine-resistant P. berghei ANKA strain. There were two control groups of animals, one group received amodiaquine alone while the other group received saline. Parasiticidal activity and survival of the animals were assessed over 21 days. RESULTS: Parasitaemia in the control animals peaked at 38% on day 9 and all animals died by day 10. The combination of amodiaquine with rifampicin 15 mg/kg body weight was the most effective of all the combinations and more efficacious than amodiaquine alone. The order of superiority of anti-malarial efficacy of the combinations was as follows; AQ + RIF 15 > AQ > AQ + ART + RIF 30 > AQ + ART + RIF 15 > AQ +RIF 30. CONCLUSION: The combination of the clinical dose of rifampicin (15 mg/kg) with amodiaquine represents a potentially valuable treatment option in management of drug resistant malaria. In addition, the role of pharmacokinetic interaction in multiple drug therapy cannot be over-emphasized.

Efficacy of artemisinin combination therapy for the treatment of uncomplicated falciparum malaria in Nigerian children.

INTRODUCTION: The development and spread of Plasmodium falciparum resistance to most commonly used antimalarials remain a major challenge in the control of malaria. Constant monitoring of drug efficacy is an important tool in establishing rational antimalarial drug policies. METHODOLOGY: A randomized comparative study was conducted at the Wesley Guild Hospital, Ilesa, Nigeria between February 2010 and September 2011 comparing the efficacy and safety of artemether-lumefantrine (Coartem) and fixed dose of artesunate plus amodiaquine (Larimal) in the treatment of uncomplicated P. falciparum malaria in children betweem 6 and 144 months of age. P. falciparum malaria parasitemia was assessed by microscopy and rapid diagnostic test. Drugs were administered according to age for three days under supervision. The primary efficacy endpoint was a day 28 PCR-corrected parasitological cure. RESULTS: A total of 182 patients were enrolled in the two treatment groups, Coartem (n = 101) and Larimal (n = 81), and tested after 28 days. In the intention-to-treat population, Coartem (n = 101) and Larimal (n = 81) had a PCR-corrected cure rate of 98% and 100% respectively, while in the per-protocol population, Coartem (n = 89) and Larimal (n = 71) both had a PCR-corrected cure rate of 100% at day 28. Although parasite and fever clearance time was faster in the Larimal group, no significant difference was observed between the two drugs. No serious adverse effects  were reported. CONCLUSION: Five years after being introduced in Nigeria, both Coartem and Larimal have been shown to be safe and highly effective in the treatment of uncomplicated P. falciparum malaria in children.

In vivo antimalarial activity of Trichilia megalantha harms extracts and fractions in animal models.

The crude methanol extracts of leaf, stem bark, root bark and stem bark fractions of Trichilia megalantha (Meliaceae) were screened for in vivo antimalarial activities in mice against a chloroquine resistant Plasmodium berghei berghei ANKA clone using the 4-day suppressive test procedure. Chloroquine diphosphate was used as the positive control. The extracts demonstrated intrinsic antimalarial property. Of all the seven extracts studied, the stem bark gave the highest activity. At 200 mg/kg of mouse, the stem bark extract had complete suppression of parasite growth (100%). Least activity was observed for the leaf extract, while the root bark had a parasite suppression of 98.4% at 800 mg/kg comparable to that of Chloroquine. Percentage suppression of parasite growth on day 4 post-infection ranged from 3.1 to 96.1% in mice infected with P. berghei and treated with extracts and fractions of T. megalantha when compared with chloroquine diphosphate, the standard reference drug which had a chemosuppression of 96.2%. At 400 mg/kg, the stem bark chloroform fraction was the most active fraction with 89.1% parasite growth suppression followed by the ethyl acetate fraction (76.4%), hexane soluble fraction (54.8%) and methanol fraction (20.5%). The mean survival time of mice that received extract ranged from 8.75 ± 0.65 to 26.0 ± 1.2 days (increased as the dose increases to 800 mg/kg), which was statistically significant, except the lowest dose (100 mg/kg) compared to the negative control group mice (9.45 ± 0.6 days). The animals that were treated with Chloroquine had mean survival time of 23.5 ± 1.2 days.

In vitro interaction of artemisinin derivatives or the fully synthetic peroxidic anti-malarial OZ277 with thapsigargin in Plasmodium falciparum strains.

BACKGROUND: Semi-synthetic artemisinin derivatives are powerful peroxidic drugs in artemisinin-based combination therapy (ACT) recommended as first-line treatment of Plasmodium falciparum malaria in disease-endemic countries. Studies by Eckstein-Ludwig and co-workers showed both thapsigargin and artemisinin specifically inhibit the sarcoplasmic reticulum Ca²âº-ATPase of Plasmodium falciparum (PfATP6). In the present study the type of interaction between thapsigargin and artemisinin derivatives as well as the ozonide OZ277 (RBx11160 or arterolane) was evaluated in parasite cultures. The latter compound is an adamantane-based peroxide and the first fully synthetic clinical candidate recently registered in India by Ranbaxy Laboratories Ltd. for anti-malarial combination therapy. METHODS: Drug interaction studies were performed using a previously described fixed ratio method and anti-malarial activity measured using the [3H] hypoxanthine incorporation assay. RESULTS: The sum 50% and 90% fractional inhibitory concentration (∑FIC50, 90) of the interaction of thapsigargin with OZ277, artemether or artesunate, against NF54 and K1 strains of P. falciparum ranged from 0.9 to 1.4. CONCLUSION: The interaction of thapsigargin with OZ277, artesunate or artemether was additive, data consistent with previous observations indicating that activity of anti-malarial peroxides does not derive from reversible interactions with parasite targets.

Efficacy of Artemisinin Combination Therapy for the Treatment of Uncomplicated Falciparum Malaria in Nigerian Children

Introduction: The development and spread of Plasmodium falciparum resistance to most commonly used antimalarials remain a major challenge in the control of malaria. Constant monitoring of drug efficacy is an important tool in establishing rational antimalarial drug policies.Methodology: A randomized comparative study was conducted at the Wesley Guild Hospital; Ilesa; Nigeria between February 2010 and September 2011 comparing the efficacy and safety of artemether-lumefantrine (Coartem) and fixed dose of artesunate plus amodiaquine (Larimal) in the treatment of uncomplicated P. falciparum malaria in children betweem 6 and 144 months of age. P. falciparum malaria parasitemia was assessed by microscopy and rapid diagnostic test. Drugs were administered according to age for three days under supervision. The primary efficacy endpoint was a day 28 PCR-corrected parasitological cure. Results: A total of 182 patients were enrolled in the two treatment groups; Coartem (n = 101) and Larimal (n = 81); and tested after 28 days. In the intention-to-treat population; Coartem (n= 101) and Larimal (n= 81) had a PCR-corrected cure rate of 98 and 100 respectively; while in the per-protocol population; Coartem (n = 89) and Larimal (n = 71) both had a PCR-corrected cure rate of 100 at day 28. Although parasite and fever clearance time was faster in the Larimal group; no significant difference was observed between the two drugs. No serious adverse effects were reported. Conclusion: Five years after being introduced in Nigeria; both Coartem and Larimal have been shown to be safe and highly effective in the treatment of uncomplicated P. falciparum malaria in children

Antitrypanosomal activity of some medicinal plants from Nigerian ethnomedicine.

Human African trypanosomiasis is a neglected tropical disease with complex clinical presentation, diagnosis, and difficult treatment. The available drugs for the treatment of trypanosomiasis are old, expensive, and less effective, associated with severe adverse reactions and face the problem of drug resistance. This situation underlines the urgent need for the development of new, effective, cheap, and safe drugs for the treatment of trypanosomiasis. The search for new antitrypanosomal agents in this study is based on ethnomedicine. In vitro antitrypanosomal activity of 36 plant extracts from 10 plant species from Nigerian ethnomedicine was evaluated against bloodstream forms of Trypanosoma brucei rhodesiense STIB 900. Cytotoxic activity was determined against mammalian L6 cells. Alamar blue assay was used to measure the endpoint of both antitrypanosomal and toxicity assays. The ethyl acetate extract of leaves of Ocimum gratissimum Linn. (Labiatae) showed the highest antitrypanosomal activity (IC(50) of 2.08 ± 0.01 µg/ml) and a high selective index of 29. Furthermore, the hexane, ethyl acetate, or methanol extracts of Trema orientalis (L.) Blume (Ulmaceae), Pericopsis laxiflora (Benth. ex Baker) Meeuwen, Jatropha curcas Linn. (Euphorbiaceae), Terminalia catappa Linn. (Combretaceae), and Vitex doniana Sweet (Verbenaceae) displayed remarkable antitrypanosomal activity (IC(50) 2.1-17.2 µg/ml) with high selectivity indices (20-80) for trypanosomes. The antitrypanosomal activity of T. catappa and T. orientalis against T. brucei rhodesiense (STIB 900) is being reported for the first time in Nigerian ethnomedicine, and these plants could be a potential source of antitrypanosomal agents.

Interaction between ciprofloxacin and chloroquine in mice infected with chloroquine resistant Plasmodium berghei: interaction between ciprofloxacin and chloroqune.

The increasing spread of chloroquine resistant malaria has intensified the search for new antimalarial treatment, especially drugs that can be used in combination. Ciprofloxacin (CFX) a fluoroquinolone commonly used to treat bacterial infections has been shown to possess significant antimalarial activity both in vitro and in vivo. Thus efforts in this study were devoted to evaluating the antimalarial activity of combination of chloroquine (CQ) with varying doses (10, 20, 40 80, 160 mg/kg body weight) of CFX in groups of 35 mice inoculated intraperitoneally with 10(7) chloroquine resistant strain Plasmodium berghei ANKA. Parasitological activity and survival of the animals were assessed over 21 days. Parasitemia in non-treated control mice peaked at 78% on day 9 and none survived by day 10. However, the combination of CQ with 160 mg/kg body weight of CFX resulted in a reduction in parasitemia between days 9 and 14 and this was significantly lower than that obtained with CQ alone or CQ combined with the lower doses of CFX (p < 0.05). In addition, the combination of CQ with 160 mg/kg CFX significantly reduced mortality in the infected animals (p = 0.0002) compared with the other treatment groups. The results from this study support the potential usefulness of CFX in combination with antimalarial drugs for the treatment of chloroquine resistant human malaria.

Antidiarrheal activity of Pyrenacantha staudtii Engl. (Icacinaceae) aqueous leaf extract in rodents.

ETHNOPHARMACOLOGICAL RELEVANCE: Pyrenacantha staudtii Engl. (Icacinaceae) is a plant which is traditionally used for the treatment of blemnorrhea, hernia, insomnia, intestinal pain and diarrhea in Nigeria. Therefore the core aim of the present study is to evaluate antidiarrheal activity of Pyrenacantha staudtii aqueous extract (PSE). MATERIALS AND METHODS: The antidiarrheal activity was evaluated using castor oil-induced diarrhea method. The effects of Pyrenacantha staudtii aqueous extract on gastrointestinal motility, intestinal transit and enteropooling were also examined in rodents. The acute toxicity effect of the aqueous extract of Pyrenacantha staudtii was also investigated. RESULTS: Pyrenacantha staudtii aqueous extract (PSE, 100-400mg/kg, p.o.) produced dose-dependent and significant (P<0.05-0.01) protection of rats and mice against castor oil-induced diarrhea, inhibited intestinal transit, and delayed gastric emptying. PSE, produced dose-dependent and significant (P<0.05-0.01) antimotility effect, caused dose-related inhibition of castor-oil-induced enteropooling in animals, comparable to atropine (1mg/kg, p.o.). Like loperamide (10mg/kg, p.o.), PSE, dose-dependently and significantly (P<0.05-0.01) delayed the onset of castor-oil induced diarrhea, decreased the frequency of defecation, and reduced the severity of diarrhea in the rodents. Compared with control animals, PSE, dose-dependently and significantly (P<0.05-0.01) decreased the volume of castor oil-induced intestinal fluid secretion, and reduced the number, weight and wetness of fecal droppings. CONCLUSION: The findings of this study indicate that PSE possesses antidiarrheal property in rats and mice. These findings confirm the ethnomedicinal use of Pyrenacantha staudtii leaf as a valuable natural remedy for the treatment, management and/or control of diarrhea.

Comparison of SYBR Green I-, PicoGreen-, and [3H]-hypoxanthine-based assays for in vitro antimalarial screening of plants from Nigerian ethnomedicine.

The standard method for in vitro antimalarial drug screening is based on the isotopic assay which is expensive and utilizes radioactive materials with limited availability, safety, and disposal problems in developing countries. The use of non-radioactive DNA stains SYBR Green I (SG) and PICO green (PG) for antimalarial screening had been reported. However, the use of the two DNA stains for antimalarial screening of medicinal plants has not been compared. Thus, this study compared SG, PG with the [(3)H]-hypoxanthine (HP) incorporation assays for in vitro antimalarial screening of medicinal plants. The 50% inhibitory concentration (IC(50)) values obtained using the three methods for antimalarial activity of medicinal plants and standard antimalarial drugs were similar. Data generated from this study suggests that the non-radioactive micro-flourimetric assay is sufficiently sensitive to reproducibly identify plant extracts with antimalarial activity from those lacking activity. The HP-based assay exhibited the most robust signal-to-noise ratio of 100, compared with signal-to-noise ratios of 7 for SG and 8 for PG. The SG-based assay is less expensive than the PG- and HP-based assays. SG appears to be a cost-effective alternative for antimalarial drug screening and a viable technique that may facilitate antimalarial drug discovery process especially in developing countries.