Open Access Original Research Article

Anti-inflammatory and Analgesic Activities of Alpinia nigra Fruit Extract in Laboratory Animals

Biplab Kumar Das, Ummul Khayer Fatema, Md. Selim Hossain, Rokshana Rahman, Kaniz Fatema

European Journal of Medicinal Plants, Page 1158-1166
DOI: 10.9734/EJMP/2014/8703

Aim: This study was aimed to evaluate the possible anti-inflammatory and analgesic properties of the ethanol extract of fruit of Alpinia nigra (Zingiberaceae).

Study Design:  Assessment of anti-inflammatory and analgesic activity.

Place and Duration of Study: Department of Pharmacy, North South University, Dhaka, Bangladesh, between June 2012 and February 2013.

Methodology: The crude extract was investigated for anti-inflammatory effect on Long Evans rats using carrageenan induced paw edema method. For anti-inflammatory study, 20 rats were divided into 4 different groups each receiving either distilled water, standard drug or the extract at the doses of 250 and 500 mg/kg body weight. The analgesic activity was evaluated by hot plate; acetic acid induced writhing method in Swiss Albino mice divided into 4 different groups (control, standard diclofenac sodium and extract at two different doses of 250 and 500 mg/kg body weight).

Results: The results of preliminary phytochemical analysis revealed the presence of alkaloids, flavonoids, tannins, glycosides in significant amounts. The present study assessed anti-inflammatory activity of its fruit extracts at a dose of 250 mg/kg and 500 mg/kg against carrageenan induced paw edema in long Evans rats. Both the extracts were able to show a dose dependent anti-inflammatory activity as compared to diclofenac sodium used as a standard. The extract elicited a highly significant (p<0.001) analgesic activity in a dose dependent manner on hot plate and acetic acid induced writhing methods.

Conclusion: The anti-inflammatory and analgesic effect of the ethanol fruit extract of A. nigra may be due to the presence of various chemical constituents especially flavonoids, tannins, alkaloids or terpenoids. These experimental findings would further establish the scientific basis of the traditional uses of the plant in the management of inflammatory conditions as well as control of pain.


Open Access Original Research Article

Effect of Magnifera indica Leaf Extract on Paracetamol-induced Hepatic Toxicity in Rats

Ezeja Maxwell, Omeh Yusuf, Ezeigbo Ihechiluru, Eze Blessing

European Journal of Medicinal Plants, Page 1167-1177
DOI: 10.9734/EJMP/2014/10114

Aims: This study evaluated the effect of methanolic leaf extract of Magnifera indica on paracetamol-induced hepatic toxicity in rats.

Study Design: Hepatic toxicity in rats was induced by oral administration of paracetamol followed by treatment with the leaf extract and evaluation of liver function parameters, lipid peroxidation activity and hematology.

Place and Duration of Study: Department of Veterinary Physiology, Pharmacology and Biochemistry, Michael Okpara University of Agriculture, Umudike, Abia State, Nigeria/ One year.

Methodology: Hepatic injury was achieved by oral administration of 2000mg/kg of paracetamol to rats. Three test doses (100, 200 and 300mg/kg) of Magnifera indica leaf extract (MILE) and a standard reference drug, silymarin (100mg/kg) were administered to the rats orally for ten days through gastric gavage. At the end of the treatment, blood was collected from the rats for liver function tests, hematology, malondiadehyde (MDA) levels and catalase activities. The effect of the extract was compared with silymarin and distilled water controls.

Results: Liver function test showed that the extract and reference drug caused various levels of significant (P=0.02 to P=0.001) reduction of Aspartate aminotransferase (AST), Alanine aminotransferase (AST), Alkaline phosphatase (ALP) and total bilirubin when compared to negative control. Hematological analysis indicated various levels of  significant increase in packed cell volume, hemoglobin concentration, Red blood cell count and White blood cell count (P=0.05-P<0.001). The MDA was also significantly (P=0.043) reduced by silymarin and MILE at the doses of 200 and 300mg/kg while there was no significant (P=0.24) changes in catalase activities of both treated and control rats. 

Conclusion: This study showed that Magnifera indica leaf extract ameliorated paracetamol-induced liver toxicity with optimum effect at 300mg/kg.


Open Access Original Research Article

Preliminary Phytochemical Screenings and Antipyretic, Analgesic and Anti-inflammatory Activities of Methanol Extract of Vernonia cinerea Less. (Fam: Asteraceae)

Md. Khairul Bashar, Mohammed Ibrahim, Irin Sultana, Md. Imran Hossain, Zakya Tasneem, Md. Ruhul Kuddus, Ridwan Bin Rashid, Mohammad Abdur Rashid

European Journal of Medicinal Plants, Page 1178-1185
DOI: 10.9734/EJMP/2014/10050

Aims: The aim of the current study was to undertake phytochemical screenings and evaluate antipyretic, analgesic and anti-inflammatory activities of the methanol extract of whole plant of Vernonia cinerea Less. (VCME).

Place and Duration of Study: The study was carried out for one year in 2012 in the Department of Pharmacy, Southern University Bangladesh, Chittagong, Bangladesh.

Methodology: For preliminary phytochemical screenings, the crude methanol extract of V. cinerea was subjected to various tests to determine the chemical nature of the extract. Antipyretic activity was assessed by the yeast-induced hyperthermia in mice. The analgesic property was evaluated by formalin-induced writhing test. Acetyl salicylic acid (ASA) was used as standard in in-vitro anti-inflammatory activity test.

Results: Results of the preliminary phytochemical screening revealed the presence of alkaloids, flavonoids and triterpenoids in the extract. In yeast-induced pyrexia, the crude extract demonstrated a significant (p=0.05) reduction of mices’ body temperature after elevation by the administration of yeast. These effects were pronounced at the 2nd and 3rd h post-treatment with the extract. VCME exhibited a dose dependent activity in analgesic activity test with 32.61% and 52.17% protection at the dose of 200 and 400mg/kg, respectively as compared to that 76.09% exhibited by standard diclofenac sodium. In the anti-inflammatory test, the crude extract at the dose of 400μg/ml showed 65.12% inhibition of protein denaturation whereas standard acetyl salicylic acid (ASA) revealed 76.74% inhibition.

Conclusion: These results revealed that V. cinerea may be used in pharmaceutical applications because of its effective pharmacological properties.


Open Access Original Research Article

Antiproliferative Effect of Portlandia Extracts and Their Fractions on Breast Cancer Cells

C. Chin, M. L. Veisaga, H. A. Priestap, T. Commock, K. Campbell, B. Jestrow, J. Francisco-Ortega, M. A. Barbieri

European Journal of Medicinal Plants, Page 1186-1199
DOI: 10.9734/EJMP/2014/10250

Aim: Rubiaceae is one of the largest families of plants and it includes the Jamaican genus Portlandia. This family has been used in several ethnomedicinal practices. Thus, in this study we investigated whether extracts of two species of Portlandia affect In vitro proliferation of breast cancer cells.

Study Design: Metastatic (MDA-MB-231 and HeLa) cells were incubated in the absence or in the presence of Portlandia extracts. A human non-metastatic (MCF-10A) and normal (ATCC PCS-600-010) epithelial cell lines were used as control.

Methodology: At the end of the study all cell lines were incubated with Portlandia extracts (-and its subfractions-) and then analyzed on proliferation, thymidine incorporation and mitogenic and survival signaling pathways on metastatic and non-metastatic cells.

Results: It was found that Portlandia methanol extract inhibited proliferation of MDA-MB-231 in a dose-dependent manner attenuating the phosphorylation of Erk1/2 and Akt1 without affecting the proliferation of MCF-10A cells. [3H] thymidine incorporation was also decreased by Portlandia methanol extract. This study suggests that Portlandia extracts selectively affected the proliferation of metastatic breast cancer cells through the modulation of Erk1/2 and Akt1 activities that play a critical role during cell survival and proliferation.

Conclusion: Portlandia leaf extracts contain active compounds, which strongly repress cancer cell proliferation in a dose-dependent manner and selectively down-regulate phosphorylation of both Erk1/2 and Akt1 activities.


Open Access Original Research Article

Adventitious Bud Formation and Plantlet Regeneration of Achyrocline satureoides-A Multipurpose Medicinal Plant

D. Kotik, P. Sansberro, C. Luna

European Journal of Medicinal Plants, Page 1200-1209
DOI: 10.9734/EJMP/2014/8800

Adventitious bud regeneration was achieved from hypocotyls, cotyledons and leaf explants of Achyrocline satureioides. Organogenesis was induced from every explant cultured on Murashige and Skoog semisolid medium (plus sucrose 30g·L-1) containing different combinations of 6-benzyladenine (BA) and a-naphtalenacetic acid (NAA) under 116mmol·m2·s-1 photosynthetic photon flux density (PPFD), photoperiod 14 h and at 27±2ºC. The regeneration was similar for every tested explant and varied between 64 and 83%. The number of buds formed per regenerative explants was similar in every treatment (5-8 shoots/explant). In order to stimulate In vitro rooting, regenerative leaves were sub cultured from the best induction medium in MS lacking plant growth regulators for the same periods. Every plantlet raised In vitro was phenotypically normal and successfully hardened to ex vitro conditions. An experimental field plot with 60-day-old in vitro regenerated plants was established.

Open Access Original Research Article

Kalanchoe pinnata - a Promising Source of Natural Antioxidants

Sarita Jaiswal, Raman Chawla, S. Sawhney

European Journal of Medicinal Plants, Page 1210-1222
DOI: 10.9734/EJMP/2014/11374

Aims: Evaluation of natural antioxidant potential of Kalanchoe pinnata leaves attributable towards its therapeutic properties.

Study Design: In vitro experiments to validate antioxidant potential in aqueous and lipid phase.

Methodology: The aqueous-alcoholic whole leaf extract designated as KPE (K. pinnata extract) was subjected to comprehensive biochemical analysis to reveal its natural strength as an antioxidative agent. In lipid protection ability assay where lipid phase (pre-emulsion) was prepared using linoleic acid with Fe2+, Fe3+ and Cu2+ as stress-inducers, it’s potential to protect against peroxyl radical induced damage in non aqueous environment was tested. Deoxy-D-ribose degradation assay in presence or absence of chelating agent (EDTA) was tested to reveal non site-specific and site-specific hydroxyl radical (OHº) scavenging potential respectively. Sodium nitroprusside based nitric oxide (NO) quenching activity and nitroblue tetrazolium reduction based superoxide radical scavenging potential were also estimated.

Results: Total phenolic content of KPE was 28.4±2 μg mg-1. In lipid protection ability assay it exhibited maximally restricted Fe2+ induced amplification of peroxyl raical (ROOº) at 10 mg mL-1. It elicited a significant (P = .05) inhibition of lipid auto-oxidation by directly scavenging peroxyl radicals. In potassium ferrithiocyanate-based reducing power assay, KPE exhibited significantly higher potency as compared to the standard synthetic antioxidant butylated hydroxy toluene (BHT), in the range of 100-2000 µg mL-1. The ability of KPE to interact at the level of generation of hydroxyl radicals was also tested with deoxy-D-ribose degradation assay that revealed a two-fold higher non site-specific OHº scavenging potential than its site-specific activity. In sodium nitroprusside based NO quenching assay KPE showed >50% quenching activity at 0.5 mg mL-1.

Conclusions: KPE is a rich source of anti-oxidative properties and has strong protective potential against oxidative stress in both aqueous and lipid phases. Hydroxyl radical scavenging assay showed KPE’s ability to scavenge free radicals is more due to its reductive potency than its metal-chelation activity attributable towards its exploration in herbal drug discovery research.


Open Access Original Research Article

Phytochemical and Anti-diarrhoeal Properties of Methanolic Leaf Extract of Maerua crassifolia Forssk

G. C. Akuodor, J. A. Ibrahim, J. L. Akpan, A. U. Okorie, B. C. Ezeokpo

European Journal of Medicinal Plants, Page 1223-1231
DOI: 10.9734/EJMP/2014/11246

Maerua crassifolia Forssk. Leaf is used in African traditional medicine for management of gastrointestinal disorders. The anti-diarrhoeal activity of the methanol extract of Maerua crassifolia leaf was investigated in rats. The phytochemical screening was also carried out. The methanol extract of Maerua crassifolia leaf dose dependently decreased intestinal propulsion of charcoal meal in rats. Maerua crassifolia also exerted significant anti-enteropooling effect in rats. A profound anti-diarrhoea activity was observed when the extract was tested in diarrhoeic rats. The frequency of defecation as well as the wetness of the faecal droppings was significantly reduced. Furthermore, the leaf extract produced 100% inhibition of castor oil-induced diarrhoea in rats. Phytochemical screening revealed the presence of alkaloids, saponins, tannins, terpenoids, flavonoids, steroids, resins and cardiac glycosides. The oral LD50 obtained was greater than 5000 mg/kg in rats. The study showed that the methanol extract of Maerua crassifolia leaf possesses anti-diarrhoeal activity and its action may be linked partly to direct inhibitory effect of the extract on the propulsive movement of the gastrointestinal tract smooth muscle.


Open Access Original Research Article

Cytotoxic, Thrombolytic and Membrane Stabilizing Activities of Swietenia mahagoni (L.) Jacq. Flower Extract

S. M. Ashikur Rahman, Maksuda Akter, Tasnuva Easmin Hira, Tasnuva Sharmin, Md. Junayed Nayeen

European Journal of Medicinal Plants, Page 1232-1239
DOI: 10.9734/EJMP/2014/11047

Aims: To investigate cytotoxic, thrombolytic and membrane stabilizing activities of methanol extract and its different Kupchan partitionates of flowers of Swietenia mahagoni.

Study Design: Evaluation of cytotoxic activity using brine shrimp nauplii, thrombolytic and membrane stabilizing activities on human RBCs.

Place and Duration of Study: Phytochemical Research Laboratory, Department of Pharmacy, School of Health Science, State University of Bangladesh, from April to September, 2013.

Methodology: The eggs of brine shrimp nauplii were hatched in artificial sea water for 24 hours. Cytotoxic activity was determined by measuring the percentage of their mortality after application of different partitionates to them. Human RBCs were obtained from 3 healthy volunteers. Thrombolytic activity was calculated by weighing the clot before and after addition of different partitionates while membrane stabilizing activity was evaluated in terms of inhibition of percentage of haemolysis of RBCs by measuring optical density in both hypotonic and heat induced conditions.

Results: The highest cytotoxic activity was achieved with the crude methanol extract (LC50 = 0.10±0.01 mg/ml) among the partitionates while vincristine sulfate, the positive control, achieved an LC50 value of 0.40±0.02 mg/ml. While investigating thrombolytic activity, the petroleum ether soluble fraction achieved the highest clot lysis activity (34.30±0.78%) compared to the standard streptokinase (70.27±1.26%). While determining the membrane stabilizing activity, in hypotonic solution induced condition, the crude methanol extract inhibited 84.71±3.25% haemolysis of RBCs whereas in heat induced condition, the aqueous soluble fraction inhibited 86.30±4.98% haemolysis of human RBCs. Here, acetyl salicylic acid (0.01mg/ml) used as reference standard showed 71.91±2.29% and 45.45±4.87% inhibition of haemolysis of human RBCs in hypotonic solution and heat induced conditions, respectively.

Conclusion: From our investigation, it can be suggested that, the flower extractives can further be studied extensively to find out their efficacy.


Open Access Original Research Article

In vitro Activity of Garlic (Allium sativum) on Some Pathogenic Fungi

Elham Abdelbasit Suleiman, Wafa Ballal Abdallah

European Journal of Medicinal Plants, Page 1240-1250
DOI: 10.9734/EJMP/2014/10132

Aim: This study was conducted to investigate the in vitro antifungal activity of garlic (Allium sativum) on some pathogenic fungi.

Study Design: This is a comparative evaluation report on garlic as an antifungal agent.

Place and Duration: Department of mycology, Veterinary Research institute, between June-October 2013.

Methodology: Samples of garlic were obtained from a local market. It was thoroughly, cleaned, peeled and pulverized. Aqueous and organic extracts of garlic were obtained by maceration and Soxhlet extractor apparatus. The methanol and petroleum ether extracts were tested against Candida albicans, Aspergillus, Curvularia and some Dermatophyte species using cup diffusion and agar incorporated methods. Diameter of   Inhibition zones of growth were measured in millimeter (mm) and expressed as Mean ±SD.

Results: The obtained results revealed that aqueous and petroleum ether extracts possess the stronger activity and a broader fungicidal spectrum against tested fungi compared to methanol extract. The study also showed that the dry coarsely- powdered garlic was found to be more potent to Candida albicans than the commercial Nystatin. 

Conclusion: The study demonstrated the potent activity of garlic against tested fungi which encourages its use as a suitable alternative drug for controlling fungal infections because it has far less risk of side-effects than most known antifungal drugs and   it can be used indefinitely in quite large amounts. Therefore, adding garlic to food (raw) or crushing and swallowing raw cloves which are cheaper is recommended as a powerful anti-fungal agent. Further purification and formulation of the garlic would give a true antifungal activity comparable to standard antibiotics.


Open Access Original Research Article

In vitro Antisickling and Radical Scavenging Activities of a Poly-herbal Formula (Drepanoalpha®) in Sickle Cell Erythrocyteand Acute Toxicity Study in Wistar Albino Rats

K. N. Ngbolua, P. T. Mpiana, D. S. T. Tshibangu, P. P. Mazasa, B. Z. Gbolo, E. K. Atibu, J. N. Kadima, F. M. Kasali

European Journal of Medicinal Plants, Page 1251-1267
DOI: 10.9734/EJMP/2014/11861

Aims: To evaluate the antisickling and radical scavenging activities and acute toxicity of indigenous nutritive formula Drepanoalpha®, produced through a bio-guided based plant selection.

Study Design: Drepanoalpha® extracts, Antisickling activity by Emmel test, Antioxidant activity by 1,1-diphenyl-2-picrylhydrazyl bleaching methods; acute toxicity on rats, determination of biological and haematological parameters.

Place and Duration of Study: Science Faculty University of Kinshasa, between January 2013 and February 2014.

Methodology: The antisickling and antioxidant activities of Drepanoalpha® were determined using Emmel and the 1,1-diphenyl-2-picrylhydrazyl bleaching methods respectively. Acute oral toxicity test was performed to determine the LD50. Liver and kidney functions, the hematological and histopathological examinations were assessed using standard techniques.

Results: Obtained results revealed that Drepanoalpha® possessesinteresting in vitro antisickling and antioxidant activities as revealed by the observed normal biconcave form of sickle erythrocyte (normalization rate >80%) and the radical scavenging activity (ED50= 0.604 ± 0.028 µg/mL). Acute toxicity assessment revealed that the medium lethal dose (LD50) is higher than 4000 mg/kg. Drepanoalpha® significantly increases the values of WBC, RBC, Hb, HCT, PLT, IDR-CV and PCT. Furthermore, this polyherbal formula significantly decreases the values of IDR-SD, P-RGC, AST and ALT (p<0.05). Both the control and treated groups displayed comparable non altered histological architecture of the liver cells.

Discussion: The mean values of biochemical markers and hematological markers of treated rats revealed that Drépanoalpha® is potentially safe indicating non-toxic effect of the phytomedicine on immune cells and blood clotting factors. Moreover, this poly-herbal formulation increases the hemoglobin rate in the all treated rats (500-4000 mg/kg bodyweight) and preserves the histological architecture of the liver cells.

Conclusion: Drepanoalpha® may increase weight gain, promote erythropoiesis and thrombopoeisis in sicklers patients. This phytomedicine could be used in the treatment of all form of anemia and may also prevent bile duct obstruction or intra-hepatic cholestasis. The results can form the basis for clinical trials in humans.