Open Access Original Research Article

Effects of Inorganic Mulches on Achillea millefolium L. Sesquiterpene Lactones

M. P. González-Hernández, R. M. Barros, R. Romero Franco

European Journal of Medicinal Plants, Page 163-173
DOI: 10.9734/EJMP/2013/2648

Aims: Sesquiterpene lactones are one of the most important groups of bioactive substances in Achillea millefolium, a perennial herb cultivated because of its medicinal interest. We sought to determine if the use of inorganic mulches for weed control in A. millefolium crops could lead to differences in their sesquiterpene lactones (SLs) content.

Study Design:  A field experiment using a randomized complete block design with three replications was conducted to determine the effect of black polypropylene, black polyethylene of 100 µm and black polyethylene of 175 µm on A. millefolium SLs content.

Place and Duration of Study: Department of Crop Production, University of Santiago de Compostela, between September 2004 and August 2005.

Methodology: LC/MS analysis of A. millefolium flower heads grown over the three different inorganic mulches and in control plots was carried out in order to determine SLs.

Results: Chromatographic analysis of A. millefolium flower heads grown over polypropylene and in control plots revealed the highest diversity of SLs, whereas those developed over black polyethylene showed the least. Polypropylene mulches were shown to be the most efficient with a 2-3 fold increase of total SLs (P < 0.001). The effects of inorganic mulches on individual SLs were of limited relevance. However, the use of black polyethylene (100 µm) resulted in an increase of   8-desacetylmatricarin (P = 0.005), a SL of biological and pharmacological interest. In contrast, concentrations of three unidentified SLs decreased in yarrow plants grown over black polyethylene.

Conclusion: Weed density did not show as a major factor influencing the total amount of SLs but growth parameters such as yarrow dry matter were inversely related.


Open Access Original Research Article

An Ethnobotanical and Floristical Study of Medicinal Plants Among the Baka Pygmies in the Periphery of the Ipassa- Biosphere Reserve, Gabon

Jean Lagarde Betti, Olga Diane Yongo, Diosdado Obiang Mbomio, Donald Midoko Iponga, Alfred Ngoye

European Journal of Medicinal Plants, Page 174-205
DOI: 10.9734/EJMP/2013/2550

Aims: This paper aims to describe the popular use of medicinal plants by the Baka Pygmies settled in the periphery of the Ipassa Reserve, analyses their relative importance and characterizes the medicinal flora.

Study Design:  Gathering data on the popular use of medicinal plants in a given area.

Place and Duration of Study: Data obtained from direct interviews conducted in August 2011 in Mekob, a village settled at 10 km to the Makokou city in the North Gabon.

Methodology: The household was considered as the sample unit. For each health problem cited, the name of the plants, the plant parts, the modes of preparation, and the modes of administration of recipes were recorded. The plants were identified in the herbarium, Libreville. Plants were characterized by their phytogeographical distribution, their morphological types, their habitats, and their modes of scattering of seeds. The relative importance of the plants was established based on the number of citations “events” occurred in the recipes.

Results: Six Baka informants with an average age of 40 years old including three men and three women interviewed. A total of 136 citations composed of 71 plant species recorded in the treatment of 24 ailments. The examination of the curve showing the evolution of the number of plants with that of informants recommends enlarging the sample as to gather the maximum of plants used by the Baka pygmies. The typical Guinean species are most represented in terms of both number of plant species (72%) and citations (61.5%).

Conclusion: Some plant species cited at least twice for the same ailment are known in the literature to possess active compounds. Further studies should be undertaken to complete the sample and to investigate the affectivity of other plants that have not yet been studied for their chemical compounds and their pharmacological activity.


Open Access Original Research Article

Phytochemical Constituents and in vitro Antioxidant Capacity of Methanolic Leaf Extract of Oxytenanthera abyssinica (A. Rich Murno)

Ibeh Bartholomew O, Ezeja Maxwell, Habu Josiah Bitrus

European Journal of Medicinal Plants, Page 206-217
DOI: 10.9734/EJMP/2013/1685

Aims: The antioxidant and phytochemical properties of leaf extracts of the Nigerian Oxytenanthera abyssinica were evaluated at different concentrations using ascorbic acid standard. This preliminary study investigates the potentially bioactive components of the plant which may renew research into its medicinal value.

Study Design: Cold extraction of the leaf parts followed by evaluation of phytochemicals and antioxidant capacity using chromatographic and spectrophotometric methods.

Place and Duration of Study: Department of Biochemistry, College of Natural and Applied Sciences, Michael Okpara University of Agriculture P.M.B.7267 Umudike Abia State, Nigeria.

Methodology: The antioxidant properties were determined using three assay models: the 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP) colorimetric test and  a modified version of in vitro antioxidant TBARS after cold extraction maceration. The activity was determined at different concentrations (10μg/ml,50μg/ml,100μg/ml,200μg/ml and 400μg/ml) of the extract and expressed as % inhibition. phytochemicals were determined by standard detection and spectrophotometric methods.

Results: The yield of the methanolic leaf extract of Oxytenanthera abyssinica was 3.92% w/w dry matter.  Steroids (steroid glycoside), alkaloids, saponins, tannins, cardiac glycosides, flavonoids, phlobatanins, anthroquinone and terpenes were detected while cyanogenic glycosides were absent. The quantitative analysis yielded (in mg/100g):  flavonoids (1.51±0.23), alkaloids (1.40±0.02), polyphenols (1.31±0.32), tannins (0.071±0.40) and saponins (1.2±0.10). Oxytenanthera abyssinica’s DPPH reduction was highest at 400μg/ml (82.10 ± 0.01%) with IC50 of 56.2μg/ml. The ferric reducing power of the extract at 400μg/ml was 61±1.52% (FRAP: 0.61) and the inhibition of lipid peroxidation measured as TBARS was 81.0 ±1.11%.

Conclusion: There is an indication that Oxytenanthera abyssinica contains important phytochemicals and an antioxidant capacity comparable with standard antioxidant compounds that may be linked to its beneficial effects on health.

Open Access Original Research Article

Intrinsic Efficacy of Novel Poly-herbal Formulation Activity on Hypoglycemic, Immune Potentiating and Antioxidant Activities in Diabetic Individual

Suman Gupta, Ragini Gothalwal, Nadeem Mohammad Khan

European Journal of Medicinal Plants, Page 218-232
DOI: 10.9734/EJMP/2013/2641

Aims: The study was conducted to assess the synergistic poly-herbal formulation for diabetic patients to determine its three characteristics reducing excessive sugar level to normal, immune-potentiating and antioxidant.

Study Design: Subjects were divided into four groups, Group I(NH), were normal healthy subjects, group II(DI), were diabetics, group III (TTD) were tolbutamide treated diabetic patients, group IV(HFTD), were diabetic patients receiving combination herbal formulation in the, dosage of 5 g. /day for 4 weeks.   

Place of Study: Diabetic patients were contacted and convinced from two Government Hospitals-Hamidia Hospital, Bhopal and K. N. Katju hospital, Bhopal, (M.P.), India.

Methodology: homogenous mixtures were obtained and encapsulated 500mg/per capsule. PMNL were isolated from blood and glucose level tests performed by auto-analyzer. Immune- potentiating activity was evaluated by different following methods like a ATPase sensitivity tests, cellular water content, cell, plasma membrane calcium content, Camp activity, Phaspholipase-C activity, contact angle measurement, NBT assay. Antioxidant activity evaluates by SOD and glutathione peroxidase methods.  

Results: Encouraging results prompt that, herbal formulation, which could be proved on excellent sugar level regulator. For efficient phagocytosis by PMNL such as membrane potential, cellular water content, calcium homeostatic, calcium messenger system, contact angle i.e. hydrophobicity measurement and finally particle internalization and Phagocytic index. result were exciting with herbal formulation since it was found to effective in correction of cell parameters related to phagocytosis and remarkable recovery in anti-oxidant  enzymes in diabetic patients.   

Conclusion: In summary, the results obtained in the present investigation demonstrated\ that the present formulation beneficial in management of diabetic complication.


Open Access Original Research Article

Anti-Hyperglycemic Activity of Raphia gentiliana De Wild. (Arecaceae)

P. T. Mpiana, T. A. Masunda, B. F. Longoma, D. S. T. Tshibangu, K. N. Ngbolua

European Journal of Medicinal Plants, Page 233-240
DOI: 10.9734/EJMP/2013/2459

Aims: Raphia gentiliana De Wild fruit is edible. The aim of this work is to evaluate the anti-hyperglycemic activity of this fruit for its use as medicinal food.

Study Design: Phytochemical analysis; extraction by lyophilzation process; measure of blood glucose level; calculation of glycemic load and index.

Place and Duration of Study: National Institute of Biomedical research (DR Congo), between October 2011 and June 2012.

Methodology: The study was performed in vivo (mice and humans). A Dose of 0.2g/Kg of Raphia gentiliana fruit extracts was administered to fasting (18 hours) hyperglycemic induced NMRI mice by oral application. The kinetics profile of the blood sugar level of the hyperglycemic induced mice was evaluated using a glucometer after one to two hours of administration. For humans, the Raphia gentiliana fruit was taken by 45 consenting individuals. Glycemia was measured by spectrophometry and the triangle surface area ratio’s method was used to calculate the glycemic and load index.

Results: The results about the anti-hyperglycemic activity in NMRI’s mice showed a significant decrease in blood sugar level. After oral application of aqueous Raphia gentiliana fruit extracts, the decrease of 27% and 56% were observed after respectively one and two hours. For human subjects, observed values of glycemic index and load were -3.1% and -1.36% respectively. These negative glycemic index and glycemic load values are excepted standard.

Conclusion: The obtained results indicate that Raphia gentiliana fruits have an anti-hyperglycemic activity and justify the use of this plant in Congolese traditional medicine.


Open Access Original Research Article

Studying the Possible Biotransformation of the Cytotoxic Diterpenoid Paclitaxel Using Jatropha Curcas Cell Suspension Culture

Sara A. Nassar, Sherweit H. El-Ahmady, Abla H. Nassar, Mohamed M. Al-Azizi

European Journal of Medicinal Plants, Page 241-253
DOI: 10.9734/EJMP/2013/2881

Aims: To establish a cell suspension culture of Jatropha curcas, family Euphorbiaceae for the biotransformation of the diterpenoid anti-cancer compound paclitaxel.

Study Design: The development and maintenance of callus lines from the leaves and petioles of J. curcas and the study of the culture growth curve followed by the inoculation of the callus with paclitaxel as the substrate and monitoring of the culture viability as well as substrate biotransformation.

Methodology: Explants from the leaves and petioles of J. curcas were prepared and placed on MS supplemented media for callus induction and maintained by regular sub-culturing until a stable callus line was achieved. Some of the callus tissue was transferred to MS liquid medium to produce cell suspension cultures. Three sets of cell suspension cultures were established for 5, 10 and 14 days respectively under the same conditions. Paclitaxel (5 mg) was administrated to each flask and incubated for 3, 6 and 10 days.

Results: A callus line of J. curcas leaves and petioles was maintained successfully over a period of two consecutive years without any change in growth rate. A stable cell suspension culture was developed using the obtained callus and maximum increase in fresh weight was reached on day 21 which was 5-6 fold over initial fresh weight. The cell suspension cultures were inoculated with the diterpenoid paclitaxel (5 mg) at different time intervals through the growth cycle phases. The incubation of paclitaxel proved that the cell culture biotransformation capability could not affect the paclitaxel molecular structure and that the applied dose of paclitaxel was not cytotoxic to the cell cultures up to 18 days of incubation.

Conclusion: This is the first report of a biotransformation trial using J. curcas cell culture and results obtained should be considered when using J. curcas cell line for terpenoid biotransformation.


Open Access Original Research Article

Phytochemical Compositions and In vitro Antioxidant Capacity of Methanolic Leaf Extract of Axonopus Compressus (P. Beauv.)

Bartholomew O. Ibeh, Ezeja Maxwell, Habu Josiah Bitrus

European Journal of Medicinal Plants, Page 254-265
DOI: 10.9734/EJMP/2013/1686

Aims: We evaluated the phytochemical contents and antioxidant capacity of the methanolic leaf extract of the Nigerian Axonopus compressus. This is a preliminary investigation to determining the active principle which may be involved in the antidiabetic mechanism of the plant.

Study Design: Phytochemicals and antioxidant capacity were determined using chromatographic and spectrophotometric detection methods of cold leaf extracts of Axonopus compressus.

Place and Duration of Study: Department of Biochemistry, College of Natural and Applied Sciences, Michael Okpara University of Agriculture Umudike Abia State, Nigeria.

Methodology: Antioxidant activities were investigated by three tests namely: 2,2-diphenyl-1-picryl hydrazyl (DPPH), Fe3+ to Fe2+ transformation (ferric reducing antioxidant power, FRAP) and a modified version of TBARS assay. These in vitro antioxidant models were carried out after cold extraction maceration. The antioxidant capacity was measured at varying concentrations (10 ~ 400 μ/ml) of the extract required to quench the free radicals by 50% (IC50) and expressed as % inhibition. Phytochemicals were determined by standard detection and spectrophotometric methods.

Results: The phytochemicals: saponin (1.2±0.1), alkaloid (2.10±0.12), tannin (0.71±0.4), flavonoid (1.92±0.13) and polyphenol (1.78±0.21) in mg/100g were strongly detected. The leaf extract was found to have a concentration dependent antioxidant activity comparable with that of ascorbic acid. Axonopus compressus’s DPPH reduction was highest at 400μg/ml (92.00 ± 0.002%) with IC50 of 52.2μg/ml. The ferric reducing power of the extract at 400μg/ml (78±1.83% [FRAP:0.92])  and  the inhibition of lipid peroxidation measured as TBARS was 92.±1.21%

Conclusion: The presence of these   phytochemicals and the high antioxidant power may explain the astringent action of the plant observed in its ethnomedicinal use especially in the treatment of diabetes. Our findings therefore, suggest that Axonopus compressus possess a strong antioxidant property that may substantiate its ethnomedicinal efficacy.


Open Access Original Research Article

Analysis of Genetic Variation among Populations of Withania somnifera (L.) in South India based on RAPD Markers

R. Udayakumar, S. Kasthurirengan, T. S. Mariashibu, B. Sudhakar, A. Ganapathi, E. J. Kim, K. M. Jang, C. W. Choi, S. C. Kim

European Journal of Medicinal Plants, Page 266-280
DOI: 10.9734/EJMP/2013/2801

Aim: The present study was carried out to analyze the genetic variations among 20 different populations of Withania somnifera (L.) Dunal collected from different habitats (locations) by RAPD analysis.

Methodology: DNA was isolated from the fresh leaf samples collected from the field by Bernatsky and Tankley method. Isolated genomic DNA was purified by phenol: chloroform: isoamyl alcohol (25:24:1) extraction mixture and then amplified by MJ themal cycler. Amplified DNA products were quantified and then subjected to RAPD analysis by the method of Williams et al.

Results: Randomly amplified polymorphic DNA (RAPD) was used to analyze the genetic variation and relationship among 20 populations of Withania somnifera collected from different part of South India, including the states of Tamilnadu, Puducherry, Kerala, Andhra Pradesh, Karnataka, Gujarat, Maharashtra and supplemented by two commercial varieties from Uttar Pradesh and Delhi. Out of 40 primers, 11 selected primers produced 96 consistent RAPD markers ranging in size from 0.2 kb to 4.0 kb; out of which 75 were polymorphic. Similarity indices were estimated using the Dice coefficient of similarity and cluster analyses were carried out on the similarity estimates using the unweighted pair-group method to produce a dendrogram using arithmetic average (UPGMA) in the NTSYSpc-verson 1.80 software. The similarity coefficient ranges from 0.53 to 0.98, suggesting that the pronounced genetic variations exist among populations of W. somnifera in South India. The cluster analysis indicates that the 20 populations of W. somnifera were divided into five major groups, regardless of geographical locations.

Conclusion: The RAPD analysis indicates existence of genetic variations in natural populations and it may influence and produce changes in phytochemical constituents of W. somnifera populations.


Open Access Original Research Article

Comparative Antimicrobial Activity and Phytochemical Analysis of Datura stramonium L. Plant Extracts and Callus In vitro

Pallavi Sharma, Ram Avatar Sharma

European Journal of Medicinal Plants, Page 281-287
DOI: 10.9734/EJMP/2013/2284

The phytochemical analysis and antimicrobial activities of in vitro grown callus and in      D. stramonium plant were studied and compared. The crude extracts from D. stramonium were analysed for moisture, starch, carbohydrate, ascorbic acid, lipid, proline, crude protein, phenols, DNA, RNA, chlorophyll and carotenoid in plant parts and callus. The phytochemical content of naturally grown plant was comparatively higher than in vitro grown callus. The antimicrobial potential of the methanolic extracts of root, stem, leaves, fruits, callus and crude metabolite rich fractions were evaluated against  Escherichia coli MTCC 1652, Staphylococus aureus MTCC 3160, Pseudomonas aeruginosa MTCC 847, Aspergillus flavus MTCC 2456, Aspergillus niger MTCC 282, Fusarium culmorum MTCC 349 and Rhizopus stolonifer MTCC 2591. The results indicated that methanolic leaf extract exhibited antimicrobial activity against S. aureus (IZ=18.2mm) and E. coli (IZ=19.8mm), P. aeruginosa (IZ=22.2mm), R. stolonifer (IZ=21.5mm), and callus exhibited antimicrobial activity against A. niger (IZ=12.1mm), F. culmorum (IZ=18.9mm) and A. flavus (IZ=12.8mm). The present study also revealed that antimicrobial activity was higher in naturally grown plant. In the metabolite rich fraction (flavonoids, phytosterols and alkaloids), greatest bactericidal and fungicidal activity was exhibited by flavonoids against P. aeruginosa (IZ=22.4mm) and A. flavus (IZ= 20.1mm).


Open Access Original Research Article

Antiepileptogenic and Anticonvulsant Actions of Dalbergia saxatilis (Hook, F.) in Sub-toxic Chemical Kindling and Toxic Convulsant Models

Omoniyi K. Yemitan, Olufunmilayo O. Adeyemi

European Journal of Medicinal Plants, Page 288-296
DOI: 10.9734/EJMP/2013/3334

Aims: The aqueous root extract of Dalbergia saxatilis (DS) is used in traditional African medicine to manage convulsions and epilepsies. This study aimed at investigating DS action against models that mimic seizure development in the neurons of epileptics, the sub-toxic dose kindling models.

Study Design: Sub-toxic doses of strychnine and picrotoxin chemical kindling models; as well as single-dose toxic bicuculline convulsive models in mice.

Place and Duration of Study: Neuropharmacology Unit Laboratory, Department of Pharmacology, College of Medicine, University of Lagos, Lagos, Nigeria, between July 2006 and March 2008.

Methodology: Strychnine kindling was produced by a 48h interval, i.m administration of 1.5mg/kg strychnine for 9 trials. The mice were treated with 200mg/kg, p.o. DS, before strychnine thus: Group I: throughout the 1st - 9th kindling; group II: During the 1st - 5th kindling; and group III: during the 6th - 9th kindling trials. Control group received distilled water instead of DS throughout the 1st - 9th kindling trials. For picrotoxin study, a subconvulsant dose of 1.5mg/kg picrotoxin was injected i.p. 3 times a week for 10 weeks, 200mg/kg of DS was administered orally before picrotoxin thus: Group I: throughout the 1st - 30th kindling trials; group II: during the 1st - 12th kindling trials; group III: during the 13th - 30th kindling trials; control group received distilled water instead of DS throughout the 30 trials. Behavioural seizures were classified for seizure stages. In another study, DS (50-200 mg/kg, p.o.) was administered to mice, 30 min. before 10mg/kg, s.c. bicuculline and onset to seizures and time to death noted.

Results: DS significantly (P=.05) retarded the development and progression of strychnine kindling, but did not reverse already reached kindled state. Moreover, DS significantly (P=.05) retarded the development of picrotoxin kindling, decreased the scoring from kindling progression and prevented convulsion in fully picrotoxin-kindled mice. A significant delay of seizure onset, with complete protection at 200mg/kg DS was produced against bicuculline seizures in mice.

Conclusion: DS may attenuate development of seizures in both GABAergic and glycinergic mechanisms and be useful in the prevention of seizures as well as neuroprotection in epileptics, justifying its use in the folkloric management of epilepsies.


Open Access Original Research Article

Antioxidant Enzymes Activity and Hormonal Changes Following Administration of Ethanolic Leaves Extracts of Nauclea latifolia and Gongronema latifolium in Streptozotocin Induced-Diabetic Rats

Grace S Effiong, Bob IA Mgbeje, Godwin O Igile, Justin I Atangwho, Eyong U Eyong, Patrick E Ebong

European Journal of Medicinal Plants, Page 297-309
DOI: 10.9734/EJMP/2013/2976

Aim of the Study: To evaluate the effects of ethanolic leaf extracts of Gongronema latifolium (G.L) and Nauclea latifolia (N.L) on antioxidant enzymes activity (GPx, SOD and CAT) and hormonal status (T3, T4, Insulin, c-peptide) in streptozotocin-induced diabetic Wistar rats.

Material and Methods: Thirty six (36) albino Wistar rats of both sexes weighing 150-250g were divided into 6 groups of 6 rats each. Groups 1, 2 and 3 received 400mg/kg body weight (b.w) of G.L, N.L and 200mg/kg b.w each of G.L and N.L respectively while group 4 received 5 iu/kg b.w of insulin subcutaneously daily for 21 days, Groups 5 and 6 served as controls (diabetic and Normal) and received placebo. Fasting blood glucose was determined at the start of the experiment and thereafter at 72 hours interval and at the end of experimental period. The animals were sacrificed and sera preparations were used for antioxidant enzymes and hormonal assays.

Results: Blood glucose in diabetic animals decreased significantly (P=.05) by 66.34%, 18.12%, 67.73% and 86.62% of initial values upon treatment with G.l, N.l, G.I plus N.I and insulin respectively. There was only a 24.44% decrease in the diabetic control. A significant decrease (P=.05) in insulin and T3 levels was observed in the diabetes-induced rats (65 and 85% respectively) compared to NC. The levels of the hormones where however significantly increased (P=.05) on treatment of the diabetic animals with G.l, N.l, G.I plus N.I and insulin.  Whereas a significant decrease (P=.05)  was observed in T4 level of DC rats compared to the NC, treatment with the leaf extracts and insulin did not result in any elevation of the hormone relative to DC. The C-peptide levels for all groups were much lower than the corresponding insulin levels, suggesting a type 1 diabetes in the diabetes-induced rats. A significant decrease (P=.05) in activity was observed for GPx and SOD in the DC group relative to NC. A combination of G.l and N.l gave a much higher reversal in activity (P<.01) when compared to treatments with individual leaf extracts. There was a significant increase (P=.05) in CAT activity in the DC animals relative to NC. This was potentiated in all treatment groups with the combination group showing a synergy in its potentiating effect.

Conclusion: There was a reversal in the level of the hormones and the activity of the antioxidant enzymes towards normal control, and comparable to the reversals by treatment with insulin, on treatment of the diabetic animals with the leaves extracts especially in combination. The results taken together indicate a synergy that makes the combination of the two plants extracts a potent antidiabetic remedy.