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2018| April-June | Volume 10 | Issue 2
Online since
April 20, 2018
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ORIGINAL ARTICLES
Antioxidant and cytotoxic potential of endophytic fungi isolated from medicinal plant
Tragia involucrata
L.
Ananda Danagoudar, Chandrashekhar G Joshi, Sunil Kumar Ravi, Heggodu G Rohit Kumar, Balenahalli Narasingappa Ramesh
April-June 2018, 10(2):188-194
DOI
:10.4103/pr.pr_137_17
Objective:
The aim of the present study was to test the antioxidant and cytotoxic and DNA protection activities of ethyl acetate extracts of endophytic fungi isolated from
Tragia involucrata
Linn. (Euphorbiaceae).
Materials and Methods:
The 1, 1-diphenyl-2-picrylhydrazyl scavenging, reducing power, and total antioxidant assay were used to evaluate the antioxidant activity. Cytotoxic activity of endophytic fungal extracts against MCF-7 and MOLT-4 cell lines was carried out using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl-2H-tetrazolium bromide method.
Results:
We have successfully isolated four endophytic fungi from
T. involucrata,
namely
Penicillium citrinum
CGJ-C1 (GenBank accession No. KT780618),
P. citrinum
CGJ-C2 (GenBank accession No. KP739821),
Cladosporium
sp
.
CGJ-D1 (GenBank accession No. KP739822), and
Cryptendoxyla hypophloia
CGJ-D2 (GenBank accession No. KT780619). The ethyl acetate extract of
P. citrinum
CGJ-C2 showed the highest antioxidant as well as cytotoxic activity among the four fungal extracts taken for the study. All the extracts showed moderate DNA protection activity.
Conclusions
: Further studies on the isolation and purification of the lead molecule will help in designing the novel therapy for different ailments associated with free radical generation.
Abbreviations used:
MCF-7: Breast Cancer Cell Line; MOLT-4: Human leukemia cell line; DPPH: 1, 1-diphenyl-2-picrylhydrazyl; MTT:3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide; DMSO: Dimethyl sulfoxide; H2O2: Hydrogen peroxide; NCI: National Cancer Institute.
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Formulation and evaluation of anti-inflammatory cream by using
Moringa oleifera
seed oil
Vidyadhara Suryadevara, Sandeep Doppalapudi, LC Sasidhar Reddivallam, Ramu Anne, Mounika Mudda
April-June 2018, 10(2):195-204
DOI
:10.4103/pr.pr_101_17
Background:
Natural oils have a variety of pharmaceutical applications. They can be used in the preparation of a formulation which is beneficial as addictives and also pharmacological agents. One of such medicinally important plants is
Moringa oleifera.
Objective:
The current investigation mainly focuses on the development of anti-inflammatory cream using
Moringa
seed oil and its pharmacological evaluation.
Materials and Methods:
The oil was extracted from
Moringa
seeds using cold pressing technique and then subjected to phytochemical screening which revealed the presence of alkaloids, glycosides, tannins, and flavonoids. Various creams were prepared with alkali saponification of free fatty acids present in the oil to get o/w type of emulsion type cream. Various physicochemical tests such as the determination of viscosity, pH, irritancy, dye test, and accelerated stability studies were performed for the prepared creams.
Results:
Of all the creams, the formulation MF
4
prepared with 500 mg potassium hydroxide was suitable for all acceptable characteristics of o/w type emulsion type of cream. The
in vitro
diffusion studies were carried out using Franz diffusion cell. The extracted oil was also subjected to various characterization studies such as Fourier transform infrared spectroscopy, gas chromatography-mass spectrometry, and high-performance thin-layer chromatography. The
ex vivo
anti-inflammatory activity was carried out using heat-induced hemolysis and protein denaturation techniques. Whereas
in vivo
anti-inflammatory activity was performed on male Albino rats using paw edema technique. A significant 70% reduction in paw edema was observed.
Conclusion:
Thus, the current research reveals the novel formulation with traditional
Moringa
oil having anti-inflammatory potency.
Abbreviations Used:
FT-IR: Fourier Transform Infra Red; GC-MS: Gas Chromatography – Mass Spectroscopy; HPTLC: High Performance Thin Layer Chromatography; MF4 - Moringa oleifera formulated cream.
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Phytochemical screening, total phenolic content, antioxidant, and cytotoxic activity of five peruvian plants on human tumor cell lines
Oscar Herrera-Calderon, Corina Alvarado-Puray, Jorge Luis Arroyo-Acevedo, Juan Pedro Rojas-Armas, Victor Chumpitaz-Cerrate, Renán Hañari-Quispe, Rita Valenzuela-Herrera
April-June 2018, 10(2):161-165
DOI
:10.4103/pr.pr_109_17
Background:
Cancer is considered a serious public health problem in the world; searching alternative treatments of medicinal plants constitute a promissory field to find new anticancer drugs.
Objective:
The aim of this study is to evaluate the phytochemical screening, total phenolic content (TPC), antioxidant, and cytotoxic activity of ethanol extracts of
Waltheria ovata
(WO),
Piper aduncum
(PA),
Myrciaria dubia
(MD),
Physalis peruviana
(PP), and
Geranium dielsianum
(GD) on human tumor cell lines.
Materials and Methods:
Phytochemical screening was assessed using chemical reactives. TPC was developed using Folin Ciocalteu reactive, and the antioxidant activity was determined against 2,2-diphenyl-1-picrylhydrazyl (DPPH), and 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radicals by spectrophotometry. The cytotoxic activity was determined on human tumor cell lines followed as: MCF-7, H-460, HT-29, M-14, K-562, and DU-145.
Results
: Phytochemical study confirmed flavonoids and phenolic compounds in all extracts. TPC was found to be the highest in WO extract (1250 mg of gallic acid equivalent/g of dried extract) rather than other extracts. The highest antioxidant activity was stablished in WO extract for DPPH and ABTS radical scavenging tests (inhibitory concentration [IC
50
] = 0.89 ± 0.01 μg/mL, IC
50
=4.20 ± 1.50 μg/mL). Ethanolic extracts (μg/mL) showed low cytotoxicity on human tumor cell lines (CI
50
>20 μg/mL) for PA, PP, GD, MD. Meanwhile, WO presented cytotoxicity on H460, MCF-7, and K562 tumor cell lines.
Conclusion:
In our findings, WO and MD extracts revealed a high antioxidant and TPC. WO exhibited better cytotoxic effect compared with 5-FU. Hence, these medicinal plants could be effective to prevent cancer disease and oxidative stress.
Abbreviations Used:
IC50: Half inhibitory concentration. TPC: Total phenolic content. DPPH: ,2-Diphenyl-1-picrylhydrazyl. ABTS: 2,2-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid). AA: Ascorbic acid.
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Therapeutic implications of ajwa dates (
Phoenix dactylifera
) in the inhibition of liver tissue alterations through the modulation of vascular endothelial growth factor and phosphatase, and tensin homolog gene
Arshad H Rahmani, Amjad A Khan, Ali Y Babiker, Masood A Khan, Mohammad A Alzohairy
April-June 2018, 10(2):151-155
DOI
:10.4103/pr.pr_119_17
Background:
Dates are a plant species in the palm family,
Arecaceae
and is used as staple food in the Middle East for thousands of years.
Aim:
The current study was undertaken to evaluate the protective role of ajwa dates extract (ADE) on carbon tetrachloride (CCl
4
)-induced hepatotoxicity.
Materials and Methods:
The study was carried out on mice mode through different groups as control group without treatment of CCl
4
, ADE and CCl
4
-treated group, and CCl
4
-treated group only.
Results:
This finding demonstrated that histological alterations including degeneration, congestion and infiltration of lymphocytes were seen in the liver tissue in CCl
4
-treated groups. However, ADE-treated groups showed protection to attenuate CCl
4
-induced liver toxicity through maintenance of architecture of hepatocytes as evident of congestion, necrosis, and degeneration was not noticed. Moreover, a few number of infiltrations of lymphocytes were noticed in ADE-treated groups. However, expression of phosphatase and tensin homolog (PTEN) and vascular endothelial growth factor (VEGF) protein was evaluated in all groups, and it was observed that PTEN was highly expressed in control group and ADE-treated group. Whereas, PTEN expression was also observed in CCl
4
-treated group, but out of eight cases, one case showed less expression of PTEN protein. The difference in expression pattern of PTEN protein in CCl
4
-treated group and ajwa-treated group was statically insignificant (
P
> 0.05). VEGF was not expressed in control and ajwa-treated group. While expression of VEGF protein was observed in CCl
4
-treated group, and it was noticed that two out of eight cases showed expression of VEGF protein.
Conclusion:
The findings supported the idea that ADE might reduce liver tissue alterations or maintenance of architecture of hepatocytes. Ajwa dates-treated group showed decreased in the VEGF expression and such angiogenesis process involve in migration and differentiation of endothelial cells as well as prevent the loss of PTEN protein expression.
Abbreviations Used:
CCl
4
: Carbon Tetrachloride, VEGF: Vascular Endothelial Growth factor, IHC: Immunohistochemistry TUNEL assay: Terminal deoxynucleotidyl transferase dUTP nick end labeling, H and E: Hematoxylin-eosin staining, ADE: Ajwa dates extract.
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Effect of UP165, a zea mays leaf extract standardized for 6-methoxybenzoxazolinone, as sleep adjunct
Mesfin Yimam, Ping Jiao, Mei Hong, Lidia Brownell, Qi Jia
April-June 2018, 10(2):156-160
DOI
:10.4103/pr.pr_148_17
Background:
Sleep is a natural phenomenon essential for rejuvenating the body, promoting good health, upholding memory, performance, and maintaining overall health. Its deprivation is linked to the increased health risk that leads to poor quality of life and negative socioeconomic consequences. While behavioral techniques, such as improving sleep hygiene, are typically the first-line of intervention, pharmaceutical drugs are frequently used as adjuncts. However, for most, besides being too expensive, the long-term use of these drugs is marred by their severe adverse side effects and crippling dependency. As a result, significant numbers of patients are always in search for a safe and efficacious alternative from natural sources.
Materials and Methods:
We evaluated the effects of UP165, a
Zea mays
(commonly known as corn) leaf extract standardized for 6-methoxybenzoxazolinone content, on sleep latency and sleep time in pentobarbital-induced mouse sleep model. The extract was orally administered at 250 mg/kg (low dose), 500 mg/kg (mid dose), and 1000 mg/kg (high dose) daily for 32 days. The immediate impact of the extract on sleep was also assessed.
Results:
Increases of 11.6 ± 0.2 (
P
= 0.008), 10.2 ± 2.4 (
P
= 0.022), and 10.5 ± 0.9 (
P
= 0.017) minutes in sleep time were observed for the 250, 500, and 1000 mg/kg UP165 treated mice compared to vehicle control, respectively. Up to 67% sleep latency incidence was observed for mice treated with UP165 compared to the 20% in the vehicle control group. UP165 showed no immediate drowsiness effect. No differences in baseline and end of study bodyweight were observed between groups.
Conclusion:
UP165 could be used as an adjunct for a sleep disorder.
Abbreviations used:
UP165-Maizinol, 6-MBOA-6-methoxybenzoxazolinone, SAMe-S-Adenosyl Methionine, GABA-gamma-aminobutyric acid, SK2 channel-Small conductance calcium-activated potassium channels.
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Antilymphoma potential of the ethanol extract and rutin obtained of the leaves from schinus molle linn.
Fernando Calzada, Jesús Ivan Solares-Pascasio, Miguel Valdes, Normand Garcia-Hernandez, Claudia Velázquez, Rosa María Ordoñez-Razo, Elizabeth Barbosa
April-June 2018, 10(2):119-123
DOI
:10.4103/pr.pr_90_17
Background:
Schinus molle
Linn. (
Anacardiaceae
) is a medicinal plant used by traditional healers in Mexican traditional medicine as antitumoral.
Objective:
This study was undertaken to obtain information that support the traditional use of the leaves from
S. molle
as antitumoral.
Material and Methods:
Antilymphoma properties of the ethanol extract of the leaves from
S. molle
(EEL
Sm
) and rutin were made on athymic CD-1 nu/nu and CD-1 mice inoculated with U-937 cell line (human leukemic monocyte lymphoma [HLML]), and for their antiproliferative effects on U-937 cell line by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Furthermore, the extract and rutin were tested for brine shrimp (BS) toxicity test. In addition, HPLC analysis was realized to known the content of rutin in the leaves from
S. molle
.
Results:
An EEL
Sm
and rutin exhibited important cytotoxic effects on U-937 cells line (IC
50
from 172.0 μg/mL and 9.6 μg/mL, respectively) and showed
in vivo
antitumoral properties on HLML in two murine models (EC
50
from 52.2 and 9.5 mg/kg to CD-1 nu/nu mice; EC
50
from 99.4 mg/kg and 6.8 mg/kg to CD-1 mice, respectively). In addition, both showed strong lethality on BS larvae (LC
50
≤ 22.2 μg/mL). The result of HPLC showed that rutin was the major constituent of EEL
Sm
.
Conclusions:
These test results support traditional medicinal use of
S. molle
as antitumoral and also suggest that both rutin and EEL
Sm
possess antitumor effect on HLML in murine models. Finally, rutin may play an important role in anticancer properties of
S. molle
.
Abbreviations used:
EEL
Sm
: Ethanol extract of the leaves from S. molle, HLML: Human leukemic monocyte lymphoma, HPLC: High pressure liquid chromatography, BS: Brine shrimp, BSLT: BS lethality test, DMSO: Dimethyl sulfoxide, MTT: 3 (4,5 dimethylthiazol 2 yl) 2,5 diphenyltetrazolium bromide.
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Antiurolithic and antioxidant activity of ethanol extract of whole-plant
Biophytum sensitivum
(Linn.) DC in Ethylene-Glycol-induced urolithiasis in rats
BR Abhirama, R ShanmugaSundaram
April-June 2018, 10(2):181-187
DOI
:10.4103/pr.pr_83_17
Background:
Biophytum sensitivum
Linn. DC is considered as one of the ten sacred plants called as “Dasapushpam” of Kerala state in India. Grounded leaves of this plant have been used conventionally as antiurolithic and diuretic. 'However, enough scientific evidences were not available about the effect of this plant as nephroprotective and antiurolithic.
Objective:
The present study was undertaken to investigate antiurolithic and antioxidant activity of ethanol extract of whole plant
B. sensitivum
Linn. DC (EEBS) on ethylene-glycol (EG)-induced urolithiasis in Wistar albino rats.
Materials and Methods:
EG 0.75% v/v in drinking water was fed to all groups, except the control group for 28 days to induce urolithiasis in rats. Groups I, II, and III served as control, toxic control, and standard Cystone groups, respectively. Animals in Group IV were administered with EEBS from 15
th
day to 28
th
day, while Group V animals were administered with EEBS from 1
st
day to 28
th
day. Several renal functional and injury markers in urine and serum were determined. Antioxidant enzyme activities were also recorded.
Results:
Co-administration with EEBS exhibited protective effect against EG-induced proteinuria, hypercalciuria, hypomagnesuria hypercalcemia, and hyperphosphatemia. Serum protein levels were significantly increased, whereas blood urea nitrogen, creatinine, and uric acid levels were significantly lowered. EEBS-treated rats significantly attenuated the aberrations in the antioxidant enzyme activities, body weight, kidney weight, urine output, and urine pH compared to toxic control animals.
Conclusion:
Hence, this study confirmed the usefulness of
B. sensitivum
as an antiurolithic and antioxidant agent.
Abbreviations Used:
EEBS: Ethanol extract of
Biophytum sensitivum
, EG: Ethylene-glycol, B.wt: Body weight, TP: Total protein, Ca: Calcium, Mg: Magnesium, BUN: Blood urea nitrogen, K.wt: Kidney weight, SOD: Superoxide dismutase, GPx: Glutathione peroxidase, GSH: Reduced glutathione, MDA: Malondialdehyde.
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Chemical profile and biological activities of the aerial parts of
Senecio acaulis
(L.f.) Sch.Bip
Rehab M S. Ashour, Abeer M El Sayed, Mohamed F Elyamany, Dina R Abou-Hussein
April-June 2018, 10(2):130-136
DOI
:10.4103/pr.pr_149_17
Background:
Genus
Senecio
is known by its phenolic constituents, terpenoids, essential oil (EO), and pyrrolizidine alkaloids. No previous reports could be traced about the phytochemical study of
Senecio acaulis
.
Objectives:
To investigate the chemical composition and biological potentiality of EO of
S. acaulis
aerial parts and to study the phytoconstituents of the plant extract and its spasmolytic activity.
Materials and Methods:
The EO was obtained by hydrodistillation and its chemical composition was analyzed by gas chromatography coupled to mass spectrometry
. In-vitro
screen of antimicrobial, antimalarial, and antileishmanial activities was determined against positive controls. Column chromatography was used to isolate the phytoconstituents from chloroform and ethyl acetate fractions; their structures were elucidated using physical and spectral methods. Spasmolytic activity was measured before and after K
+
-induced contractions on isolated rabbit jejunum. DNA-fingerprint was established by RAPD-PCR technique using 12 primers.
Results:
The study of EO revealed the detection of 22 compounds representing 81.08% of the oil composition. The major constituents were D-limonene (13.32%), β-pinene (11.54%), and sabinene (10.79%). Eight compounds were isolated from the plant extract and identified as β-amyrin, β-sitosterol, lupeol, oleanolic acid, β-amyrin-3-O-β-glucopyranoside, isorhamnetin 3-O-β-glucopyranoside, isoquercitrin, and quercitrin. The oil exhibited moderate antimalarial activity against chloroquine-sensitive and chloroquine-resistant strains of
Plasmodium falciparum
. The oil showed a significant antimicrobial activity against methicillin-resistant
Staphylococcus aureus
and
Cryptococcus neoformans
and weak antileishmanial activity. In isolated rabbit jejunum, the ethanol extract produced a relaxation of spontaneous and moderate effect against high K
+
(80 mM)-induced contractions. Amplification of DNA yielded 87 RAPD fragments.
Conclusion:
Senecio acaulis
(L.f.) Sch.Bip essential oil can be used as antimicrobial agent against
Staphylococcus aureus
and Cryptococcus neoformans. In addition, the spasmolytic activity of its ethanolic extract suggests its incorporation in antidiarrheal preparations. Further clinical trials are required to evaluate these effects on humans. Identification of twenty two compounds in its essential oil, isolation of eight compounds for the first time as well as authentication of the plant via DNA finger print may play an important role in its chemotaxonomic classification.
Abbreviations Used:
EO: Essential oil, GC: Gas Chromatography, GC-MS: Gas Chromatography-Mass Spectrophotometry, ETOAc: Ethyl acetate, SEM: Standard error mean, RRI: Relative retention indices, SI: Selectivity Index, D6: chloroquine sensitive Plasmodium falciparum, W2: chloroquine resistant Plasmodium falciparum. SI: selectivity index, IC50, IC90: concentration that affords 50 and 90% inhibition, respectively, 1H-NMR: Proton Nuclear magnetic resonance, 13C-NMR: Carbon-13 Nuclear magnetic resonance.
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Effect of pomegranate (
Punica granatum
) seed oil on carbon tetrachloride-induced acute and chronic hepatotoxicity in rats
Duygu Yaman Gram, Ayhan Atasever, Meryem Eren
April-June 2018, 10(2):124-129
DOI
:10.4103/pr.pr_122_17
Background:
Carbon tetrachloride (CCl
4
) is one of the most widely used Hepatotoxin that is known to induce oxidative stress and causes hepatic damage by the formation of reactive free radicals in laboratory animals.
Objective:
This study aims to investigate the hepatoprotective role of pomegranate seed oil (PSO) on histological structure, some biochemical parameters and lipid peroxidation on CCl
4
-induced acute and chronic liver injury induced rats.
Materials and Methods:
The study material comprised 80 male Wistar albino rats. They were divided into two study groups including 40 rats for acute and 40 rats for chronic hepatotoxicity induction by CCl
4
. Hematoxylin and eosin staining was used to evaluate degree of steatosis, inflammation, necrosis, and fibrosis semiquantitatively. Blood serum aspartate transaminase, alanine transaminase, and alkaline phosphatase enzyme activities and glucose, triglyceride, total cholesterol, high-density lipoprotein-cholesterol, low-density lipoprotein-cholesterol, total protein, albumin and liver malondialdehyde, and nitric oxide levels were measured.
Results:
All control and only PSO given animals liver showed normal histological architecture, but in the acute CCl
4
-treated animals, an intensive macro and microvesicular steatosis, mononuclear inflammatory cell infiltrations in portal area and parenchyma, and necrotic alterations; in the chronic CCl
4
-treated group, additionally to acute findings mild-to-severe fibrosis with lobulation formation were observed.
Conclusion:
The results suggest that administration of PSO has partially ameliorative effects on biochemical and lipid peroxidation parameters in acute period, but it has no effect on the recovery of liver tissue damage or histopathological changes and biochemical parameters induced by CCl
4
in chronic period.
Abbreviations Used:
PSO: Pomegranate seed oil; CCl
4
: Carbon tetrachloride; CCl
3
: Trichloromethyl; MDA: Malondialdehyde; NO: Nitric oxide; ROS: Reactive oxygen species; IM: Intramuscular; ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; ALP: Alkaline phosphatase; TP: Total protein; HDL: High density lipoprotein; LDL: Low density lipoprotein.
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Anti-inflammatory effect of procumbenoside B from
Justicia spicigera
on lipopolysaccharide-stimulated RAW 264.7 macrophages and zebrafish model
Rosa Martha Perez Gutierrez, Jose Maria Mota Flores, Adriana Maria Neira Gonzalez
April-June 2018, 10(2):218-224
DOI
:10.4103/pr.pr_97_17
Background:
Justicia spicigera
is widely used in the Mexican traditional medicine for the treatment of inflammation.
Objectives:
The purpose of this study was to investigate the anti-inflammatory effect of procumbenoside B (PB) from
J. spicigera
on pro-inflammatory mediators.
Materials and Methods:
Lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages and Zebrafish model were used to assess the potential anti-inflammatory effects of PB; its structure was elucidated on the basis of spectroscopic data. The production of inflammatory mediators such as interferon-β, prostaglandin E2, inducible nitric oxide synthase (iNOS), nuclear factor-kappa B (NF-κB) p65, interleukin-6 (IL-6), IL-1β, IL-12, cyclooxygenase (COX-2), tumor necrosis factor-α, and anti-inflammatory IL-10 was measured using enzyme-linked immunosorbent assay. NO production was measured using Griess reagent.
Results:
In LPS-induced-inflammatory response in RAW264.7 macrophage cells, PB strongly inhibited secretion of all pro-inflammatory mediators test and increased the production of IL-10 and blockade of NF-κB. In addition, PB suppressed LPS-stimulated nitric oxide and reactive oxygen species generation in a zebrafish model.
Conclusion:
These results indicated that the anti-inflammatory effects of PB may be attributed to the downregulation of iNOS and COX-2 through the suppression of NF-κB signaling pathway in RAW264.7 macrophages.
Abbreviations used:
4-Amino-5-methylamino-2',7'-difluorofluorescein diacetate (DAF-FM-DA), Cyclooxygenase-2 (COX-2), 2,7-dichlorofluorescein (DCH-DA), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), Dimethylsulphoxide (DMSO), Dulbecco's modified eagle's medium (DMEM), Fetal bovine serum (FBS), Granulocyte-macrophage colony-stimulating factor (GM-CSF), Interferon-β (IFN β), Interleukin-6 (IL-6), Interleukin-8 (IL-8), Interleukin-1B (IL-1b), Layer chromatography (TLC), Lipopolysaccharide (LPS), Matrix metalloproteinases-3 (MMP-3), Matrix metalloproteinases-13 (MMP-13), Nitric oxide (NO), Nitric oxide synthase (iNOS), Nuclear factor-kappa B (NF-βB), Prostaglandin E2 (PGE2), Reactive oxygen species (ROS), Tumor necrosis factor-β (TNF-β).
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Detection of estrogenic, antiestrogenic, and drug synergistic activities of seven commercially available fruits by
In Vitro
reporter assays
Paramita Basu, Dinu Dixon, Sherin Varghese, Camelia Maier
April-June 2018, 10(2):137-142
DOI
:10.4103/pr.pr_111_17
Background:
Fruits are known to possess antiosteoporotic and anticancer properties in part due to their estrogenic and antiestrogenic activities.
Objective:
In this study, estrogenic, antiestrogenic, and drug synergistic activities of seven commercially available fruits were evaluated.
Materials and Methods:
A steroid-regulated transcription system in
Saccharomyces cerevisiae
containing a human estrogen receptor alpha expression plasmid, and a β-galactosidase gene reporter plasmid was employed for the estrogenic, antiestrogenic, and drug agonistic studies.
Results:
California table grape extract showed the highest estrogenic activity. The estrogenic activities of other extracts ranked as follows: blackberry, red raspberry, strawberry > blueberry > jackfruit, black raspberry. The transcriptional activities of the combination estradiol-fruit extracts (FEs) (400E equivalents) ranked as follows: blueberry (95.9%), blackberry (86.2%), black raspberry (88.9%), and California table grape (81.5%) > jackfruit (72.2%), and red raspberry (73.2%) > strawberry (60.7%). Black and red raspberry extracts showed the highest synergistic activities with 4-hydroxytamoxifen (4-OHT). Black and red raspberry extracts in combination with 4-OHT lowered the estradiol activity by 74.9% and 73.9%, respectively. The highest synergistic activity with nafoxidine (NAF) was displayed by red raspberry extract. Together, NAF and red raspberry extract lowered estradiol activity by 77.9%. Fold changes were calculated for drug synergistic activities of FEs, and they ranged from 1.3 to 15.3 for 4-OHT and 1.5–17.4 for NAF, respectively.
Conclusions:
The active compounds in the FEs studied may be useful in enhancing the antiestrogen activities of chemotherapy drugs and be used as chemopreventive agents for patients at high risk of estrogen-induced cancers.
Abbreviations Used:
4 OHT: 4 hydroxytamoxifen, Abs: Absorbance, CAA-glucose medium: Casamino acid glucose medium, E: Estrogen/Estradiol, ER: Estrogen receptor, ERα: Estrogen receptor alpha, FE: Fruit extract, LBD: Ligand binding domain, MU: Miller Unit, NAF: Nafoxidine, RQC: Resveratrol, quercetin, and catechin.
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High-performance liquid chromatography-fingerprint analyses,
In vitro
cytotoxicity, antimicrobial and antioxidant activities of the extracts of two cestrum species growing in Egypt
Sami M Nasr, Mosad A Ghareeb, Mona A Mohamed, Nehal M Elwan, Abd El-Wanes Anter Abdel-Aziz, Mohamed S Abdel-Aziz
April-June 2018, 10(2):173-180
DOI
:10.4103/pr.pr_145_17
Background
: Cancer diseases and microbial resistance are serious health disorders associated with oxidative stress and infectious diseases. Their risks can be reducing via using polyphenols-rich plants.
Methodology
: Different solvent extracts from two
Cestrum
species (
Cestrum nocturnum
and
Cestrum elegans
) were evaluated for their biological and chemical activities. Also, the chemical profiles of the most promising extracts were investigated via high-performance liquid chromatography (HPLC)-fingerprint analyses.
Results
: The tested extracts showed weak to moderate cytotoxicity against Vero cell line with IC
50
values ranged from 133.67 μg/ml to 57.634 μg/ml. The only noncytotoxic extractive fraction was the dichloromethane extract of
C. elegans
leaves with an IC
50
value of 204.732 μg/ml, while the most toxic extract was the ethyl acetate extract of
C. elegans
flowers with an IC
50
value of 19.22 μg/ml. The antimicrobial activity results revealed that the
n
-BuOH extract of
C. nocturnum
was the most active against four tested microbial strains with inhibition zones (10–13 mm). Also, the water and
n
-BuOH extracts of
C. elegans
leave exhibited moderate activities with inhibition zones (7–9 mm), while for
C. elegans
flowers both of water and methanol extracts showed strong activities (9–14 mm). In the 2,2'-diphenyl-1-picrylhydrazyl assay, the most active fraction was EtOAc with IC
50
values of 100.52 μg/ml and 64.40 μg/ml for
C. elegans
leaves and flowers respectively, while for
C. nocturnum
the most active fraction was methanol with an IC
50
value of 161.16 μg/ml, all relative to 7.60 μg/ml of ascorbic acid. HPLC-fingerprint analyses revealed that the major identified compounds in the ethyl acetate extract of
C. elegans
flowers are caffeic acid, coumaric acid, vanillin, and rutin, while for the
n
-butanol extract of
C. nocturnum
leaves are coumaric acid and vanillin.
Conclusion
: The obtained results revealed that the two species can be used as natural sources of antioxidant compounds with low cytotoxic effect on the mammalian cell line.
Abbreviations Used:
HPLC: High-performance liquid chromatography; IC
50
: Median inhibitory concentration; DPPH: 2,2'-Diphenyl-1-picrylhydrazyl; ATCC: American Type Culture Collection; ECACC: European Collection of Animal Cell Cultures; HEPES: 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; GM: Growth medium; EDTA: Ethylene diamine tetra acetic acid; MM: Maintenance media; ELISA: Enzyme-linked immuno-sorbent assay; G
+ve
: Gram-positive; G
−ve
: Gram-negative; IP: Inhibition percentage; RP-HPLC: Reversed phase-high performance liquid chromatography; DAD: Diode array detection; NCI: National Cancer Institute; DCM: Dichloro methane.
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Evaluation of protective effects of hydroalcoholic extract of
Cassia fistula
Linn. Pod on pancreas in streptozotocin-induced diabetic rats
Ram Niwas Jangir, Gyan Chand Jain
April-June 2018, 10(2):205-212
DOI
:10.4103/pr.pr_95_17
Background:
Diabetes mellitus (DM) is associated with oxidative stress. Medicinal plants and herbs are the rich sources of antioxidants which ameliorate oxidative stress-induced diabetic complications and could play an important role in the management of diabetes.
Objective:
The present study aimed to evaluate the protective effects of 70% ethanolic extract of
Cassia fistula
pod on pancreas in streptozotocin (STZ)-induced diabetic rats.
Materials and Methods:
Diabetes was induced in male Wistar rats by single intraperitoneal injection of STZ (60 mg/kg b.wt.). The diabetic rats were administered orally with
C. fistula
pod extract at three different doses (100, 250, and 500 mg/kg b.wt./day) for 60 days. The results were compared with standard drug glibenclamide (5 mg/kg b.wt./day) treated rats. Relative pancreatic weight and serum insulin level were determined. Histopathological changes and oxidative stress parameters, i.e., lipid peroxidation (thiobarbituric acid reactive substance [TBARS]) and antioxidative defense markers (superoxide dismutase, catalase, glutathione, and ascorbic acid), in the pancreas were investigated.
Results:
Oral administration of
C. fistula
pod extract (100, 250, and 500 mg/kg b.wt./day) or glibenclamide in diabetic rats significantly improved serum insulin level, total protein concentration, relative pancreatic weight, and mean diameter of islets of Langerhans as compared to diabetic control rats. Furthermore, treatment with extract also reduced TBARS levels and improved the levels of antioxidant markers in the pancreas. The histomorphological picture of the pancreas showed marked restoration of islets morphology. These results were comparable with glibenclamide.
Conclusions:
The results of the present study showed that
C. fistula
pod extract possesses significant antidiabetic activity though enhanced insulin secretion, improvement of antioxidative status of pancreas, and preservation of the integrity of pancreatic islets.
Abbreviations used:
b.wt.: Body weight; CAT: Catalase; DM: Diabetes mellitus; DNA: Deoxyribonucleic acid; GSH: Glutathione; ROS: Reactive oxygen species; SOD: Superoxide dismutase; STZ: Streptozotocin; TBARS: Thiobarbituric acid reactive substances.
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Protective effect of struthanthus marginatus on ethanol-induced gastric damage in mice
Sônia Maria de Farias Freire, Rômulo Vieira Silva, Tábata Maira Freitas Morais, Rachel Melo Ribeiro, Selma do Nascimento Silva, Maria do Socorro de Sousa Cartágenes, Iracelle Carvalho Abreu, Maria Nilce de Sousa Ribeiro, Danilo da Cruz Centeno, Luce Maria Brandão Torres, Marilene Oliveira da Rocha Borges, Antonio Carlos Romão Borges
April-June 2018, 10(2):143-150
DOI
:10.4103/pr.pr_76_17
Background:
Our research group previously characterized the antioxidant and gastroprotective effects of
Struthanthus marginatus
(Loranthaceae), a medicinal herb used in Brazil as a healing agent.
Objective
: The aim of this study is to evaluate the chemical composition of aqueous extract of
S. marginatus
(AESm), as well as the mechanisms underlying its gastroprotective and ulcer healing properties using different protocols in mice.
Materials and Methods:
Gas chromatography-mass spectrometry and liquid chromatography/electrospray ionization-mass spectrometry-mass spectrometry/diode array detection analyses to evaluate the chemical composition of AESm were conducted. The antisecretory activity (basal or stimulated) was determined using the pyloric ligature method. The gastroprotective action of nitric oxide and sulfydryl groups (–SH groups) were evaluated using ethanol-induced gastric ulcer model. The healing ability was evaluated using an acetic acid-induced chronic ulcer.
Results:
Chromatographic analyses of AESm permitted to identify several compounds, including 3-
trans
-caffeoylquinic acid (3-
trans
-CGA), quercetin, and kaempferol as the major constituents. Oral treatment of animals with AESm (500 mg/kg/day) reduced the severity of ethanol-induced gastric damage similar to omeprazole and in a more pronounced manner than 3-
trans
-CGA. Such effect was significantly reduced in animals pretreated with N
ω
-nitro-L-arginine methyl ester. In addition, AESm inhibited gastric acid secretion in pylorus-ligated mice stimulated with histamine or pilocarpine similar to atropine or cimetidine, respectively. A decrease in acetic acid-induced gastric ulcers similar to that promoted by cimetidine was also observed.
Conclusion:
The results show that
S. marginatus
is rich in flavonoids and that these compounds contribute directly to the gastroprotective and ulcer healing effects of this herb. The inhibition of gastric secretion is the possible gastroprotective mechanism.
Abbreviations Used:
AESm: Aqueous Extract of S. marginatus; 3-trans-CGA= 3-trans-caffeoylquinic acid.
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Antidepressant effect of
Hedyotis corymbosa
extract in olfactory bulbectomy rats
Anil T Pawar, Gayatri D Gaikwad, Bhanudas S Kuchekar
April-June 2018, 10(2):213-217
DOI
:10.4103/pr.pr_118_17
Background:
Hedyotis corymbosa
Linn. (family:
Rubiaceae
) has been used in the Indian indigenous system of medicine, Ayurveda, for treatment of various health aliments.
Objectives:
The present study was undertaken to evaluate the antidepressant activity of methanolic extract of whole plant of
H. corymbosa
(MHC) in olfactory bulbectomy rats.
Materials and Methods:
MHC was prepared and standardized to gallic acid by high-performance thin-layer chromatograph method. Effects of 14 days oral treatment of fluoxetine (30 mg/kg) and three doses of MHC (50, 100, and 200 mg/kg) were evaluated on olfactory bulbectomy (OBX)-induced alterations in behavioral and biochemical parameters in rats.
Results:
MHC treatment reversed the OBX-induced behavioral abnormalities such as increased exploratory activity in open-field paradigm and decreased feeding latency time in novelty-suppressed feeding test. Serum cortisol levels were restored near to normal by the MHC treatment. Further, treatment of MHC prevented the OBX-induced decline of brain levels of serotonin and nor-adrenaline in dose-dependent manner.
Conclusion:
MHC prevents behavioral and neurochemical changes in OBX-induced depression in rats. These results demonstrate the antidepressant effect of
H. corymbosa
and support its folklore claim.
Abbreviations Used:
ANOVA: one-way analysis of variance; CPCSEA: Committee for the Purpose of Control and Supervision of Experiments on Animals; ELIZA: Enzyme-linked immunosorbent assay; HPTLC: High-performance thin-layer chromatography; MHC: Methanolic extract of Hedyotis corymbosa; NA: Noradrenaline; OBX: Olfactory bulbectomy; SEM: Standard error of mean; TLC: Thin layer chromatography; 5-HT: 5-hydroxytryptamine; 5-HIAA: 5-hydroxyindoleacetic acid.
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SHORT COMMUNICATIONS
Gas chromatography-mass spectrometric determination of components of leaves of
Aegle marmelos
and
Psidium guajava
and seeds of
Nigella sativa
and correlation with
In vitro
antioxidant activity
CV Jayachandran Nair, Sayeed Ahamad, Washim Khan, Varisha Anjum, Rajani Mathur
April-June 2018, 10(2):230-235
DOI
:10.4103/pr.pr_93_17
Objective:
The aim of this study is to analyze the volatile components of the leaves of
Aegle marmelos, Psidium guajava
, and seeds of
Nigella sativa
, using gas chromatography-mass spectrometric (GC-MS) and correlate with their
in vitro
antioxidant activity.
Materials and Methods:
Plant material was analyzed for heavy metal content (HMC), pesticide residue (PR), and the presence of microbes/fungi. Following hydrodistillation, the volatile components (AM, PG, and NS) were subjected to GC-MS analysis and
in vitro
antioxidant assay (2,2-diphenyl-1-picrylhydrazyl).
Results:
The plant material passed the test for HMC, PR, microbial, and fungal contamination. Using GC-MS, the number of compounds identified in AM, PG, and NS were 62, 46, and 58, respectively. The antioxidant activity of AM, PG, and NS was 98.538%, 98.955%, and 97.755%, respectively.
Conclusion:
GC-MS-based methods can be successfully utilized for phytochemical profiling and standardization of plant material.
Abbreviations Used:
AM: Volatile oil of Aegle marmelos Correa.; GC-MS: Gas chromatography-mass spectrometry; HMC: Heavy metal content; NS: Volatile oil of Nigella sativa Linn.; PG: Volatile oil of Psidium guajava Linn.; PR: Pesticide residue.
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2,043
107
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Dual anti-cholinesterase activity of ajoene by
In silico
and
In vitro
studies
Shivani Kumar, Sayan Chatterjee, Suresh Kumar
April-June 2018, 10(2):225-229
DOI
:10.4103/pr.pr_140_17
The two major forms of cholinesterase enzymes found in the mammalian brain are acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE). BuChE usually found mainly in glial cells and neuron in normal physiological condition, whereas AChE found near nerve synapse and axons, both are involved in the breakdown of acetylcholine (ACh) in the brain. The dual inhibition of these enzymes is considered as a promising strategy for the treatment of a neurological disorder such as Alzheimer's disease, senile dementia, ataxia, and myasthenia gravis. The objective is to study the dual anticholinesterase activity of ajoene using
in silico
and
in vitro
methods. The anticholinesterase activity of ajoene was evaluated using Ellman's assay, and molecular docking was performed on Schrödinger suite software. The present study demonstrated ajoene ([
E
,
Z
]-4, 5, 9-trithiadodeca-1, 6, 11-triene-9-oxide) inhibited both AChE and BuChE in a concentration-dependent manner. The IC
50
value of ajoene was 2.34 mM for AChE and 2.09 mM for BuChE. Kinetic studies showed mixed noncompetitive inhibition of AChE and uncompetitive inhibition of BuChE. Molecular docking studies revealed that ajoene interacts hydrophobically with catalytic residues of AChE while in case of BuChE the interaction is through noncatalytic binding site residues. Ajoene exhibits dual inhibitory activity against both AChE and BuChE enzymes.
Abbreviations Used:
AChE: Acetylcholinesterease; BuChE: Butyrylcholinesterase; AD: Alzheimer's disease; Ach: Acetylcholine; ChAT: Choline acetyltransferase; ATChI: Acetylthiocholine iodide; BuChI: Butyrylcholine iodide; DTNB: 5,5-Dithiobis-2-nitrobenzoic acid; PDB: Protein data bank; RMSD: Root mean square deviation; OPLS: Optimized potentials for liquid simulations; ADME: Absorption, distribution, metabolism, excretion; DADS: Diallyl disulfide; QPlogPo/w: Predicted octanol/water partition coefficient; donorHB: Number of hydrogen bond donors; accptHB: Number of hydrogen bond acceptors; MW: Molecular weight of the compounds.
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2,054
91
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ORIGINAL ARTICLES
Inter simple sequence repeat analysis of genetic diversity and relationship in four egyptian flaxseed genotypes
Abeer A El Sayed, Shahira M Ezzat, Saber H Mostafa, Saeid Z Zedan, Essam Abdel-Sattar, Nebal El Tanbouly
April-June 2018, 10(2):166-172
DOI
:10.4103/pr.pr_126_17
Background
: Flaxseed is a highly important industrial and medicinal plant worldwide.
Objective
: To use inter simple sequence repeat (ISSR) technique for making unique fingerprint for the four newly produced genotypes of flax in Egypt.
Materials and Methods
: The genetic diversity among four promising Egyptian flax (
Linum usitatissimum
L.) genotypes was premeditated by means of polymerase chain reaction-based ISSR markers. The phenotypic variation among the four flax genotypes, namely, promising strains 533/39/5/3 (F1), S.402/3/3/7 (F2), S.421/3/6/4 (F3), and S.11 (F4) was studied during the two successive winter seasons of 2014/2015 and 2015/2016 in randomized complete block design through four replications.
Results
: The promising strain (F3) surpassed the other flax genotypes regarding seed yield/feddan, oil yield/feddan, and oil percentage. Twelve ISSR primers were used for the genetic examination yielding 139 loci, of which 31 were polymorphic. The middling number of amplified loci and the middling number of polymorphic loci per primer were 11.6 and 2.6, correspondingly, while the percent of loci polymorphism ranged from 0.0% to 58.0% with a middling of 21.4% crosswise all the flax genotypes. The more informative primers were GAC (GATA)
4
and (GATA)
4
GC, while the less informative were (AC)
8
T and (GT)
8
G. Unweighted pair group method with arithmetic mean derived dendrogram clearly discriminated the flax genotypes in three clusters. The Jaccard's similarity coefficient along with the genotypes ranged from 0.91 to 0.95.
Conclusion
: This study identified S. 421/3/6/4 (F3) strain to be the mainly assorted genotype and recommended its use in propagation programs and for upward mapping populations.
Abbreviations Used:
CTAB: N-cetyl-N,N,N-trimethylammonium bromide; EDTA: ethylenediaminetetraacetic acid; ISSR: Inter Simple Sequence Repeat; PCR: polymerase chain reaction; RAPDs: random amplified polymorphic DNAs; RCBD: Randomized Complete Block Design; UPGMA: Unweighted Pair Group Method with Arithmetic Mean; SSR: simple sequence repeat.
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