Open Access Original Research Article

The Antidepressant-like Effect of Hyperbrasilol B, A Natural Dimeric Phloroglucinol Derivative is Prevented by Veratrine, a Sensitive-Voltage Na+ Channel Opener

Fernanda Bossemeyer Centurião, Satchie Sakamoto, Ana Cristina Stein, Liz Girard Müller, Pietro Maria Chagas, Gilsane Von Poser, Cristina Wayne Nogueira, Stela Maris Kuze Rates

European Journal of Medicinal Plants, Page 1268-1281
DOI: 10.9734/EJMP/2014/7702

Aims: South Brazilian Hypericum species are a source of dimeric structures, constituted of filicinic acid and phloroglucinol moieties, which present antidepressant-like effects mediated by monoaminergic neurotransmission in rodents. Here, we show that hyperbrasilol B, a phloroglucinol derivative from Hypericum caprifoliatum, presents antidepressant-like activity in mice forced swimming test (FST). The aim of this study was to determinate if Na+ channels are important to the antidepressant-like effect of hyperbrasilol B and also verify the effect of this compound on Na+, K+ ATPase activity in the cerebral cortex and hippocampus of mice.

Methodology: We assessed the effects of veratrine, a Na+ channel opener on antidepressant-like effect of hyperbrasilol B by using mice FST. Veratrine (0.06 mg/kg) and hyperbrasilol B (10 mg/kg) were given i.p. 60 and p.o. 30 min, respectively, before the test. In another batch of experiments different groups of mice were treated with hyperbrasilol B 10 mg/kg, p.o. (Single administration or once a day during 3 days). Two hours after the acute or after the last of the three treatments, the brain structures were removed for measuring Na+, K+ ATPase activity.

Results: Veratrine was able to prevent the anti-immobility effect of hyperbrasilol B on the FST, suggesting that its antidepressant-like effect might be due to Na+ influx modifying properties. Animals treated for 3 consecutive days with hyperbrasilol B presented a significant increase in the hippocampus Na+, K+ ATPase activity. The acute treatment was ineffective.

Conclusion: Alterations in the Na+ gradient may be implicated in the antidepressant-like effect of hyperbrasilol B.


Open Access Original Research Article

In vitro Anthelmintic Activity of Bidens pilosa Linn. (Asteraceae) Leaf Extracts against Haemonchus contortus Eggs and Larvae

Mbogning Tayo Gertrude, J. Wabo Poné, Komtangi Marie Claire, Yondo Jeannette, M. Ngangout Alidou, Mpoame Mbida

European Journal of Medicinal Plants, Page 1282-1292
DOI: 10.9734/EJMP/2014/10421

Haemonchus contortus is a major health and welfare problem for small ruminants, responsible for economic losses through reduced productivity and increased mortality. The in vitro efficacy of Bidens pilosa was determined against this gastrointestinal nematode (GIN). Fresh eggs, embryonated eggs and larvae (L1 and L2) were incubated at room temperature in infused aqueous extract, macerated aqueous and ethanolic leaf extract of B. pilosa at concentrations of 0.625, 1.25, 2.5, 3.75 and 5 mg/ml for 48, 6 and 24 hours, respectively. Distilled water and 1.5% Tween 80 were used as negative controls. They did not affect development of eggs and larvae whereas extracts showed a concentration dependent activity eventhough aqueous extracts exhibited a weak activity on the different developmental stages of H. contortus compared to ethanolic extract. Ethanolic extract was more potent on larvae than on eggs. It inhibited 92.5±7.5% and 67.4±7.4% egg embryonation and egg hatch at 5 mg/ml, with IC50 values of 2.1 mg/ml and 3.3 mg/ml respectively and induced 100±0% and 89.8±3.2% L1 and L2 larvae mortality at 5 mg/ml with LC50 values of 1.8 and 1.96 mg/ml respectively. The overall findings of the current study indicated that the evaluated medicinal plant in occurrence B. pilosa possess potential anthelmintic effect and further in vivo and toxicity evaluation are indispensable to validate its use as anthelmintic for the control of GIN.


Open Access Original Research Article

Effects of “Lesser Known” Leafy Vegetables (Vitex doniana and Corchorus oletorius) on the Oxidative Stress Indices of Albino Rats

N. Nwachukwu, E. E. J. Iweala, H. O. Asoluka

European Journal of Medicinal Plants, Page 1293-1301
DOI: 10.9734/EJMP/2014/6396

Aim: To evaluate the effects of two “lesser known” leafy vegetables- Vitex doniana and Corchorus oletorius on the oxidative indices of Albino Rats.

Study Design: Forty eight rats, mean body weighty 351.83±1.39g were grouped into four to represent a control, and three diet groups. Dried powdered form of the vegetables was mixed with the normal rat chow in the ratio of 1:4 and pelleted before feeding to the rats. Feeding lasted for a total of three (3) months. The first analysis was done within two weeks, and thereafter repeated every two weeks throughout the study.

Place and Duration of Study: The research work was done at Biochemistry laboratory of the Federal University of Technology, Owerri and National Root Crop Research Institute, Umuahia. The study lasted for a period of three months and two weeks (104 days).

Methodology: Clean uninfected leaves of the samples were selected and sun dried to constant weight before grinding with a milling machine. The resulting powdered form was used to formulate the experimental diet with the normal rat chow in the ratio of 1:4 as shown in the text. Enzyme activities were determined according to standard methods as referenced in the text. Malonyladehyde and vitamin C contents were also determined according to standard methods.

Results: Values of malonyladehyde, Vitamin C and activities of catalase significantly (P≥0.05) increased when the sample vegetables were administered to the rats as compared with the control which received no vegetables. However, the activities of peroxidase decreased also significantly as compared with the control. However, only the increase in the values of indices determined were sustained throughout the period of study.

Conclusion: The studied vegetables may possess antioxidant components which may play important role in the management of diseases associated with oxidative stress.


Open Access Original Research Article

Stability of Active Constituents of Hops (Humulus lupulus) Strobiles and their Ethanolic Extracts during Storage

Daniel Gagnon, Chitra Wendakoon, Robert Smith, Jeremy Leker

European Journal of Medicinal Plants, Page 1302-1312
DOI: 10.9734/EJMP/2014/11250

Aims: The purpose of this study was to evaluate the stability of three major active constituents (humulones, lupulones and xanthohumol) in dried hops (Humulus lupulus) strobiles (whole and ground) as well as their ethanolic extracts during storage.

Methodology: A comparative study of humulones, lupulones and xanthohumol levels of H. lupulus strobiles during storage was carried out. Dried whole strobiles and cryogenically ground dried strobiles stored at -15ºC  as well as ethanol extracts of the strobiles prepared using different ethanol concentrations (10%, 30%, 50%, 70%, and 95%) and stored at room temperature, were analyzed by HPLC to quantify each constituent. These hops samples were analyzed immediately after preparation, and then one year and two years later to determine the concentrations of the constituents.

Results: HPLC analysis indicated that the amount of all three constituents in the ground strobiles and in the ethanol extracts decreased gradually during the storage period. The 10% and 30% ethanol extracts had very low amounts of constituents initially and were practically devoid of constituents at the end of two years. The 50% ethanol extract contained considerable amounts of humulones and xanthohumol, and low levels of lupulones initially, but lost substantial amounts over time. The 70% and 95% ethanol extracts showed higher levels of all three constituents, while the 95% H. lupulus ethanol extract contained the highest constituent levels throughout the experimental period. The ethanol content of the extract had a direct correlation to the constituent levels; the higher the ethanol level, the higher the initial and subsequent constituent levels.

Conclusion: Both dried hops and ethanol extracts lose active components over storage time. When preparing extracts, at least 70% ethanol is necessary to extract the highest levels of three bioactive constituents and to retain them over a two-year period. Ethanol concentration is a critical factor to be considered in hops extraction process.


Open Access Original Research Article

In vitro Antimicrobial Activity of the Flower Buds of Eugenia caryophyllata

Felix Charles Mills-Robertson, Gloria Adjapong, Williams Walana

European Journal of Medicinal Plants, Page 1313-1323
DOI: 10.9734/EJMP/2014/11438

Aims: This study investigated the antimicrobial effect of aqueous and ethanol extracts of the flower buds of Eugenia caryophyllata (Myrtaeae) against a wide range of bacteria and yeasts cells isolated clinically from patients.

Methodology: The agar diffusion method was used to establish the antimicrobial activity and the zones of inhibition caused by the extracts. The antimicrobial effects of 16% and 32% aqueous and ethanol extracts of Eugenia caryophyllata were investigated against 111 pathogenic bacteria and yeasts cells. The microbes used consisted of 11 Proteus mirabilis, 20 Salmonella typhi, 15 Pseudomonas aeruginosa, 18 Escherichia coli, 19 Staphylococcus aureus, 12 Klebsiella pneumoniae, and 16 Candida albicans species.

Results: The ethanol extracts inhibited the growth of all the microbes employed in the study with inhibition zones ranging from 8.00±0.00 mm to 24.00±0.00 mm. The aqueous extracts however exhibited different degrees of antimicrobial activity with zones of inhibition ranging from 6.00±0.00 mm to 13.33±0.29 mm.

Conclusion: Our study concludes that the aqueous and ethanol extracts of the flower buds of Eugenia caryophyllata have relatively good antimicrobial activity against a wide range of medically important pathogenic bacteria and Candida albicans in vitro.


Open Access Original Research Article

The Effect of Different Drying Methods on the Phytochemicals and Radical Scavenging Activity of Ceylon Cinnamon (Cinnamomum zeylanicum) Plant Parts

Darfour Bernard, Asare Isaac Kwabena, Ofosu Daniel Osei, G. Achel Daniel, S. Achoribo Elom, Agbenyegah Sandra

European Journal of Medicinal Plants, Page 1324-1335
DOI: 10.9734/EJMP/2014/11990

Aim: The research aimed at ascertaining whether the different drying methods and plant parts have effect on the radical scavenging activity and phytochemical properties of cinnamon as an herb/spice.

Methodology: Fresh samples of the cinnamon (Cinnamomum zeylanicum) was collected at the Aburi Botanical Gardens, Ghana. Some of the samples were sun, oven, room and freeze dried. The dried and fresh samples were extracted with methanol and water and the extract analyzed.

Results: Only the sun dried samples had the total phenolic and total flavonoid been degraded compared to the fresh sample. The flavonoid and phenolic contents and the DPPH radical scavenging activity were significantly expressed in different amounts in the root, stem, leaf and seed.

Conclusion: Generally, the drying influenced the phytochemical contents which are major contribution to the radical scavenging activity of the cinnamon.

Open Access Original Research Article

Polyphenolic Compounds with Anti-Ages Activity from Three Clusiaceae Plants

Joseph J. Magadula, Zakaria H. Mbwambo, Julia Gatto, Séverine Derbré, David Guilet, Pascal Richomme

European Journal of Medicinal Plants, Page 1336-1344
DOI: 10.9734/EJMP/2014/10812

Aim: This study focused on finding molecules with inhibitory effects on Advanced Glycation End-products (AGEs) formation from Tanzanian some Clusiaceae plant species

Study Design: Field study and Laboratory experimental tests.

Place and Duration of Study: Institute of Traditional Medicine, Muhimbili University of Health and Allied Sciences, P.O. Box 65001, Dar es Salaam, Tanzania and PRES LUNAM, Université d’Angers, EA 921 SONAS, 16 Bd Daviers, 49045 Angers, France, between June 2011 and July 2013.

Methodology: Three Clusiaceae plant species (Garcinia semseii, G. volkensii and Allanblackia ulugurensis) were collected and dried in the field with the assistance of a botanist. Extraction and concentration of plant samples to obtain crude extracts were done in the laboratory following standard procedures. The isolation of the phenolic compounds was carried out by using normal phase column chromatography as well as High-performance liquid chromatography (HPLC). The isolated compounds were tested for anti-AGE activity using the in vitro automated assay.

Results: Two polyphenolic compounds exhibiting phloroglucinol moieties [e.g. polyprenylated benzophenones, such as guttiferone F, 2 (18 mg)] or biflavonoids [such as morelloflavone, 1 (22mg)] were isolated and identified from A. ulugurensis and G. volkensis respectively. The results further indicated that compound 1 is an excellent inhibitor of AGE formation exhibiting an IC50 values of 78 and 64 µM at wavelength of 370/440 (vesperlysines-like AGEs) and 335/385 (pentosidine-like AGEs) respectively.

Conclusion: Plants belonging to the Clusiaceae family commonly used in Tanzanian traditional medicine need to be considered as a potential source of molecules exhibiting pharmacological activities such as anti-AGE activity. Morelloflavone (1) and other biflavonoids prove to be very good anti-AGE compounds using our automated screening assay. Hence, our automated in vitro assay allows a fast, effective and quite inexpensive screening of natural compounds and can therefore be applied to high throughput screening projects.


Open Access Original Research Article

Investigation of Antioxidant Potential of Methanolic Extract of Swertia chirata Buch. Ham.

Laxmi Ahirwal, Siddhartha Singh, Manish Kumar Dubey, Vandana Bharti, Archana Mehta

European Journal of Medicinal Plants, Page 1345-1355
DOI: 10.9734/EJMP/2014/8933

Aim: To evaluate the in vitro antioxidant potential and total phenolic contents of the methanolic extract of Swertia chirata.

Place and Duration: Department of Botany, Dr. H. S. Gour University (HSGVV), Sagar, between February 2010 to July 2010.

Methodology: The plant material (aerial part) was subjected to defatting with petroleum ether then successively extracted with methanol. Total phenolic contents of methanolic extract was determined using the Folin-Ciocalteau reagent method while in vitro antioxidant potential was evaluated by using DPPH, hydroxyl radical, nitric oxide radical scavenging as well as ferric reducing power assays.

Results: The total phenolic content in 1 mg of methanolic extract of S. chirata was equivalent to 4.5 µg Catechol. The IC50 value of the DPPH method, hydroxyl radical and nitric oxide radical scavenging activity was 222.74±0.19, 307.93±0.10 and 870.55±0.20 µg/ml respectively. When these IC50 value compared with that of standard drug Butylated hydroxy anisole (BHA) the result obtained was as follows: FRSA-SCM>BHA (p=0.000246); HRSA-SCM<BHA (p=0.000507); NORA-SCM<BHA (p=2.22614). These results showed that the S. chirata methanolic extract exhibited significant free radical DPPH scavenging activity and Hydroxyl radical scavenging activity while it exhibited non-significant Nitric oxide radical scavenging activity. In ferric reducing power various concentrations (100, 250 and 500 µg/ml) of methanolic extract of S. chirata showed absorbance 0.013±0.31, 0.156±0.12 and 0.298±0.14. Phytochemical screening showed the presence of phenolic compounds such as flavonoids and tannins which may be responsible for the activity.

Conclusion: Methanolic extract of Swertia chirata showed significant antioxidant activity which suggest the extract may act as a natural antioxidant agent offering effective protection from free radicals.


Open Access Original Research Article

Development of Simple, Cost Effective Protocol for Micropropagation of Tylophora indica (Burm f.) Merill., an Important Medicinal Plant

Pooja Patel, Rajani Nadgauda

European Journal of Medicinal Plants, Page 1356-1366
DOI: 10.9734/EJMP/2014/10801

Aims: The present studies were initiated to develop a cost effective protocol for micropropagation, as a mean for conservation of medicinal plant- Tylophora indica (Burm f.) Merill.

The plant is threatened and needs immediate conservation, therefore, the study was undertaken with following objective:

  • In vitro multiplication of Tylophora indica using nodal axillary bud proliferation and through organogenesis of callus.

Study Design: For all experiments ten replicates were used per treatment and all the experiments were repeated three times. Data have been presented as Mean ± Standard deviation.

Place and Duration of Study: Department of Plant Cell and Molecular Biology, Indian Institute of Advanced Research (IIAR), Gandhinagar, Gujarat, India.

Methodology: For in vitro plant regeneration, micropropagation and organogenesis techniques were used. For micropropagation, surface sterilized nodal explants were inoculated on different shoot inducing media and further multiplication was obtained. Root containing shoots were transferred to pot containing pre autoclaved mixture of soil and soilrite. For organogenesis, surface sterilized leaf explants were inoculated on different types of callus inducing media.

Results: All the nodal explants were sprouted at a very high frequency, i.e. 98% and sprouted buds elongated up to 8cm on three different media. Dissected explants grew further and average height of shoots reached 9.5cm±0.80cm within 30 days. Interestingly, root formation was observed on the same media; so that the best media was 0.4mg L-1 BA and 0.1mg L-1 Kn for both initiation as well as for multiplication. For organogenesis, the fragile callus was observed on media containing 2mg L-1 2,4-D and 0.1mg L-1 Kn. Green pigmented calli were transferred to MS media, where it regenerated in to shoots and roots, simultaneously

Conclusion: The protocol of micropropagation through axillary bud proliferation described here is very simple, repetitive and cost effective, which can be easily utilized for commercial cultivation. On shoot multiplication media, root formation observed, thereby making the process is one step; which is very easy to follow.


Open Access Original Research Article

Antimicrobial Activity of Selected Medicinal Plants of South-Eastern Nigeria on Pseudomonas species Expressing Extended Spectrum Beta Lactamase (ESBL)

Ikegbunam Moses Nkechukwu, Anagu Linda Onyeka, D. Nwakile Calistus, A. Afunwa Ruth, Esimone Charles Okechukwu

European Journal of Medicinal Plants, Page 1367-1377
DOI: 10.9734/EJMP/2014/8491

Aim: To evaluate the antibacterial activity of selected medicinal plants from South-Eastern Nigeria against ESBL producing Pseudomonas species.

Study Design:  Agar well diffusion assay for determination of sensitivity and Agar dilution method for determination of MIC were used.

Place and Duration of Study: Department of Pharmaceutical Microbiology and Biotechnology, Faculty of Pharmaceutical Sciences, Nnamdi Azikiwe University, Awka, Anambra State, an Igbo speaking South-Eastern State in Nigeria, between February 2010 and october 2010.

Methods: The antibiograms of ten (10) ESBL producing Pseudomonas species to selected antibiotics as well as the antibiograms of these isolates against methanol leaf extracts of Anthocleista djalonensis A Chev. (Loganiaceae) (MLEA) (Igbo name- Uvuru or Ayuu), Zapoteca portoricensis H. M. Hem. (Fabeceae/mimosidea) (MLEZ) (Igbo name- Ayuu), Gongrenema latifolium Benth.  (Asclepiadaceae) (MLEG) (Igbo name- Utazi) and Psidium guajava Linn.  (Myrtaceae) (MLEP) (Igbo name- Gova), using doses of 3000.00 to 21.87 mg/ml using agar disk diffusion and agar well diffusion assays respectively were determined. The MIC of the plant extracts in comparison with that of gentamicin were also evaluated using the agar dilution method.

Results: All the ESBL producing Pseudomonas spp. were multi-drug resistant (IZD = 0) but, were all sensitive to imipenem. Only two strains of Pseudomonas monteilli were sensitive to MLEG with sensitivity decreasing with increasing concentrations of the MLEG. All the ESBL producing Pseudomonas spp were sensitive to MLEZ and MLEP. Also, sensitivity decreased with increasing concentration of the MLEZ and MLEP. MLEA showed no antimicrobial activity against the tested ESBL producing Pseudomonas spp. Gentamicin, with an MIC of 0.00015 μg/ml, was more active than the plant extracts. The MLEP was the more active with an MIC of 1 - 4.37 mg/ml, than MLEZ and MLEG which had MICs of 150 and 75 mg/ml respectively. Active constituents of these plant extracts especially that of Psidium guajava, may thwart the emerging resistance to carbapenems.

Conclusion: Development of a complex mixture of the active constituents or single active constituent(s) of Psidium guajava as antimicrobial agent(s) that will be effective against ESBL producing Pseudomonas species.