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2021| January-March | Volume 17 | Issue 5
Online since
June 10, 2021
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ORIGINAL ARTICLES
Thin layer chromatography-mass spectrometry bioautographic identification of free radical scavenging compounds and metabolomic profile of
Carica papaya
linn. fruit and seeds using high-performance thin-layer chromatography, gas chromatography-mass spectrometry and ultra-performance liquid chromatography-mass spectrometry
Asna Khan, Sultan Zahiruddin, Mohammad Ibrahim, Parakh Basist, Gaurav , Rabea Parveen, Shahid Umar, Sayeed Ahmad
January-March 2021, 17(5):21-28
DOI
:10.4103/pm.pm_326_20
Objectives:
Carica papaya
Linn. a member of the Caricaceae family, is a tropical fruit, rich in various secondary metabolites owing to its antioxidant, anti-inflammatory, anti-diabetic, and antihelmintic properties. The study was carried out to investigate the metabolic profiling and free radical scavenging compounds in
C. papaya
fruit's pulp, peel and seeds through thin-layer chromatography-mass spectrometry (TLC-MS) bioautography.
Methodology:
Aqueous, hydroalcoholic and alcoholic extracts of fruit pulp and peel were prepared along with the hydroalcoholic and hexane extract of seeds. These were subjected to total phenol, flavonoid and free radical scavenging estimation. Qualitative and quantitative high-performance thin-layer chromatography analysis of the best extracts was performed followed by TLC-MS bio autography assay for the detection of free radical scavenging compounds.
Results:
C. papaya
peel was noted to contain the highest phenol and flavonoid content, but the seeds showed better free radical scavenging activity. The hydroalcoholic extracts of pulp, peel, and seeds examined through TLC-bioautography showed the presence of chlorogenic acid, ellagic acid, quercetin, β-sitosterol, linoleic acid, and iso-oleic acid as potent free radical scavenging compounds. Liquid chromatography-mass spectrometry analysis showed the presence of β-carotene, lycopene, and β-cryptoxanthin in pulp and peel along with other carotenoids and benzyl isothiocyanate, linoleic acid, oleic acid, and methyl palmitate were the major compounds detected in seeds through gas chromatography-mass spectrometry analysis.
Conclusion:
This study has revealed that
C. papaya
fruit and seeds possess potent free radical scavenging compounds. Seeds which make up the waste material may be utilized in cosmetic industries as they signify rich antioxidants.
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292
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Immunomodulatory and cytotoxic properties of natural triterpenoids isolated from
Grewia flavescens Juss
Gihan O M. ELhassan, Sakina Yagi, M Ahmed Mesaik, Syam Mohan, Hassan A Alhazmi, Mohammed Al-Bratty, Marai M Al-Amri, Asaad Khalid
January-March 2021, 17(5):9-14
DOI
:10.4103/pm.pm_235_20
Background:
Grewia flavescens
Juss (
Malvaceae
) is a species that is distributed throughout semi-arid and sub-humid tropical areas of Africa, Saudi Arabia, Yemen, and India. It is used in various traditional herbal practices for the treatment of various disease conditions.
Objectives:
The present investigation was undertaken to examine the immunomodulatory and cytotoxic properties of natural triterpenoids isolated from the stem bark of
G. flavescens
.
Materials
and
Methods:
The immunomodulatory activity was measured using oxidative burst chemiluminescence and phytohemagglutinin stimulated T-cell proliferation assay. The cytotoxicity was measured using MTT assay. One new triterpenoid (29 [30]-lupene-3, 20-diol; 3 β-form, 29-Aldehyde [4]) in addition to four known ones; lupeol (1), ß-sitosterol (2), betulin (3), and ß-sitosterol glucopyranoside (5), have been isolated from the stem bark of
G. flavescens
and identified using Fourier-transform infrared spectroscopy, ultraviolet, and NMR techniques.
Results:
Chemiluminescence experiments showed that no compound exerted an inhibitory effect on reactive oxygen species production. However, 29 (30)-lupene-3, 20-diol; 3 β-form, 29-Aldehyde (4) significantly repressed the cell proliferation activity with an IC
50
of 8.7 μg/mL, whereas, ß-sitosterol glucopyranoside (5) and ß-sitosterol (2) showed moderate inhibitory activities (IC
50
of 16.7 and 23.6 μg/mL, respectively) compared to the standard drug prednisolone (0.2 μg/mL). Betulin (3) revealed the highest cytotoxicity activity toward Hela cancer cells, followed by 29 (30)-lupene-3, 20-diol; 3 β-form, 29-Aldehyde (4), and ß-sitosterol (2) with IC
50
11.10, 16.5 and 21.51 μg/ml respectively.
Conclusion:
The present investigation supports the traditional use of this plant in treating various diseases and it has a good potential for future studies based on animal model based, which could bring more mechanistic facts about this plant's activity.
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1,300
188
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The traditional medicine bojungikki-tang increases intestinal motility
Hyo Eun Kwon, Jeong Nam Kim, Min Ji Kwon, Jong Rok Lee, Sang Chan Kim, Joo Hyun Nam, Byung Joo Kim
January-March 2021, 17(5):1-8
DOI
:10.4103/pm.pm_507_20
Background:
Bojungikki-tang (BJIT) is a traditional formula used to treat Gastrointestinal (GI) diseases.
Objectives:
We investigated the GI motility functions
in vivo
and the pacemaker potential in interstitial cells of Cajal (ICCs)
in vitro
by BJIT.
Materials
and
Methods:
Intestinal transit rate (ITR) and serum levels of GI hormones were investigated in mice. ICC-induced pacemaker potential was evaluated using the electrophysiological method.
Results:
ITR values and the level of motilin significantly increased after treatment with BJIT. The BJIT-induced ITR increase was related to the increase in the expression of a c-kit. BJIT induced the pacemaker potential depolarizations and the frequency decrease of ICCs. Pretreatment with methoctramine resulted in the inhibition of BJIT-induced depolarization of the pacemaker potential. However, BJIT-induced effects were retained in the presence of 1,1-dimethyl-4-diphenylacetoxypiperidinium iodide. Furthermore, thapsigargin pretreatment resulted in the inhibition of BJIT-induced effects. Moreover, BJIT blocked both transient receptor potential melastatin 7 and calcium-activated chloride (transmembrane protein 16A) channels.
Conclusion:
These results indicate that BJIT can be considered a good medicine for controlling GI motility.
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1,262
208
1
Camptothecin: An anticancer drug from
Pestalotiopsis microspora
Mh458929 – An endophytic fungus isolated from an ethnopharmacologically important medicinal plant
Cordia dichotoma
G. forst
Madhankumar Dhakshinamoorthy, Kannan Kilavan Packiam
January-March 2021, 17(5):87-95
DOI
:10.4103/pm.pm_417_20
Background:
Endophytic fungi that live asymptomatically inside the plant tissues have novel bioactive metabolites exhibiting a variety of biological activities, especially against cancer.
Cordia dichotoma
G. Forst. play a significant role in traditional medicines and therapeutics. Leaves and bark have been used as anti-inflammatory and anticancer agents.
Objectives:
Isolation, screening, and
in silico
toxicity evaluation of camptothecin (CPT) from the endophytic fungus
Pestalotiopsis microspora
MH458929.
Materials
and
Methods:
Endophytic fungus was isolated from leaves of
C. dichotoma
collected from Sathyamangalam Tiger Reserve forest (STRF), Tamil Nadu. The wild strain was identified by 18S rDNA sequencing. Modified potato dextrose broth was used as a screening medium for the presence of CPT. CPT was analyzed by high-performance liquid chromatography and electrospray ionization–mass spectrometry (ESI-MS). Compounds identified by ESI-MS from fungal extract were further studied for their
in silico
toxicity study against
Daphnia magna, Tetrahymena pyriformis, Pimephales promelas,
and
Rattus
sp. Bioaccumulation factors, developmental toxicity, and mutagenicity were studied by the quantitative structure–activity relationship model – Toxicity Estimation Software Tool.
Results:
Endophytic fungus P. microspora produced a maximum yield of 0.691 mg/L of CPT. CPT derivatives were identified at m/z of 349.10, 363.08, and 389.41 through ESI-MS analysis.
In silico
toxicity study revealed that compounds were of Category D and hence considered nontoxic to higher organisms. However, compounds showed high toxicity for lower organisms, with toxicity order
D. magna > T. pyriformis > P. promelas >
rat.
Conclusion:
The present study is the first report to screen, isolate, and analyze the CPT's
in silico
toxicity and its derivatives from endophytic fungus
P. microspora
from STRF. Further
in vitro
and
in vivo
studies are recommended to utilize CPT and its derivatives in pharmaceuticals.
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153
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Quality Analysis of
Manjishta (Rubia cordifolia
L.) with respect to pharmacognostical and high-performance thin-layer chromatography profile of the genuine sample: A cross-sectional market sample study
Vivek Palengara, CM Harinarayanan, Pratibha P Nair
January-March 2021, 17(5):45-53
DOI
:10.4103/pm.pm_394_20
Background:
The genuineness of Ayurvedic herbs causally determines the effectiveness of Ayurvedic treatment protocols. Research and subsequent evidence-based medicinal practices fundamentally dependant on appropriate identification and standardization of specific herbs are used for health-care purposes. Definite deficits that prevail in this regard in the form of adulteration, substitution, and compromised quality standards are the reasons why many scientific communities and health-related organizations question Ayurvedic sciences. This study aims at raising the very timely subject matter of genuine drug collection based on a model of Pharmacognostical and high-performance thin-layer chromatography (HPTLC) profiling of an Indian medicinal herb "
Manjishta"'
(
Rubia cordifolia
L.), a profoundly marketed Ayurvedic drug.
Objectives:
To compare the six market samples of
Manjishta
(Pharmacognostical and with HPTLC profiling) with the genuine root and stolon.
Materials and Methods:
The market samples of Manjishta from 6 districts of Kerala were collected, and these samples were compared with the genuine root and stolon (morphology, histology, and with HPTLC profiles).
Results:
Morphological and anatomical evaluation of the market samples was similar with that of the original stolon of
Rubia cordifolia
. HPTLC profiling yielded entirely different peaks in specific samples when compared with that of the genuine stolon.
Conclusion:
The collected market samples of Manjishta from different districts of Kerala were almost similar to the stolon of
Rubia cordifolia
L. A clear standard operative procedure should be prepared for medicinal plant part collection with respect to source plant maturity for Manjishta or as a matter of fact any other herb and thus the HPTLC profiles should be redefined.
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1,135
155
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Genotoxic assay of silver and zinc oxide nanoparticles synthesized by leaf extract of
Garcinia livingstonei
T. Anderson: A comparative study
Azharuddin B Daphedar, Siddappa B Kakkalameli, Govindappa Melappa, Tarikere Chandrashekharappa Taranath, Chandrashekar Srinivasa, Chandan Shivamallu, Asad Syed, Najat Marraiki, Abdallah M Elgorban, Ravindra Veerapur, Sharangouda S Patil, Shiva Prasad Kollur
January-March 2021, 17(5):114-121
DOI
:10.4103/pm.pm_536_20
Background:
Green synthesis of metal and metal oxide nanoparticles (NPs) using plant extract performs a significant role as it is a promising alternative to the conventional chemical method in nanotechnology. Aims: In this paper, we report an environmentally benign method for the synthesis of silver nanoparticles (AgNPs) and zinc oxide nanoparticles (ZnONPs) using leaf extract of Garcinia livingstonei, and their mitotic activities were investigated using the root tip of Cicer arietinum. Objectives: The as-prepared NPs were characterized by ultraviolet-visible spectroscopy, infrared spectroscopy (FT-IR), X-ray diffraction (XRD), atomic force microscopy (AFM), and high-resolution transmission electron microscopy (HR-TEM). Analysis of FT-IR spectrum revealed that certain functional groups behaved as reducing and stabilizing agents in the formation of nanostructures. The crystalline nature of the AgNPs and ZnONPs was confirmed by XRD analysis. The size and shape of the as-obtained materials were found using HR-TEM analysis and were in the range of 5–65 nm and 38–94 nm for AgNPs and ZnONPs, respectively. Further, the root cells of C. arietinum were treated with both AgNPs and ZnONPs in different concentrations (5, 25, 50, and 100 μg/ml) for 24 h at the interval of 3, 6, 12, and 24 h along with distilled water as control. Results: The study clearly indicated that the AgNPs and ZnONPs showed an inhibitory effect on the cell division in root tip cells and caused a decrease in their mitotic index (MI) values. The reduction in MI in AgNPs is more evident than that of ZnONPs when compared to control. Aberrations in chromosomal behavior such as micronucleus, sticky chromosomes, bridges, multipolar anaphase, laggard, and c-metaphase were also observed.
Conclusion:
From the results, it is evident that the percentage of MI is inversely proportional, and chromosomal aberrations (CAs) are directly proportional to the concentration and duration of exposure.
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1,042
164
1
Gas chromatography-mass spectrometry fingerprint and
in vitro
cytotoxic studies of
Rubus steudneri
leaf fractions against Michigan cancer foundation-7 breast cancer cell line
Raghavendra Lakshmana Shetty Hallur, Chaitanya V. N L. Motamarri, Prashith Kekuda T. Ramamoorthy, Chetan D Murthy, Ravikumar Patil H. Siddappa, Vijayananda N Bramhanakonda
January-March 2021, 17(5):54-62
DOI
:10.4103/pm.pm_324_20
Aim:
Rubus steudneri
Schweinf. (
Rosaceae
) is one of three
Rubus
species that grow in Ethiopia. Studies of this species have been restricted to
in vitro
antioxidant, antidiabetic, and nutritional evaluation. Until today, no identification has been made of its phytochemical fingerprints, resulting in an unclear picture of its phytochemical constituents. This study investigated the phytochemical composition of
R. steudneri
and its
in vitro
cytotoxicity against Michigan Cancer Foundation-7 (MCF-7) breast cancer cell lines and Vero cell lines.
Materials
and
Methods:
The leaf powder was subjected to liquid–liquid fractionation using
n
-hexane, chloroform, ethyl acetate, methanol, and water as solvents and the obtained fractions were subjected to flash chromatography and
in vitro
cytotoxicity studies in MCF-7 cell lines at concentrations from 1 to 1000 μg/mL, using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay.
Results:
The chloroform fraction proved to be the most promising of the bioactive fractions, with 50% of cytotoxicity inhibition for concentrations of 10 μg/mL on MCF-7 cell lines and 125 μg/mL on Vero cell lines. Two important drug leads, namely butyl isobutyl phthalate and 2-pyrrolidinone 5-(cyclohexylmethyl), were identified in chloroform fraction using gas chromatography
-
mass spectrometry (GC-MS) and may be responsible for the
in vitro
cytotoxicity. Flow cytometer results indicated that the chloroform fraction arrests cell cycle in the sub-G1 phase at a concentration of 100 μg/mL on MCF-7 cell lines, which proves that metabolites in this fraction may belong to the apoptotic population.
Conclusion:
In this study, butyl isobutyl phthalate and pyrrolidinone 5-(cyclohexylmethyl), which have significant cytotoxic effects and cell cycle arrest, are extracted. Further in-depth research is in progress to prove the anticancer activity of
R. steudneri
in search of new leads for anticancer drugs.
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978
172
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Callus induction and shoot regeneration from the immature flower bud of
Caesalpinia bonducella
and its antileptospiral potential by
in vitro
and
in silico
analysis
SR Santosh Kumar, Chandrashekar Srinivasa, Chandan Shivamallu, Kollur Shiva Prasad, Sushma Pradeep, Asad Syed, Ali Bhakali, Amar Shankar, Sharanagouda S Patil, P Ashwini, D Chandan, R Triveni, Govindappa Melappa, V Krishna
January-March 2021, 17(5):38-44
DOI
:10.4103/pm.pm_331_20
Background:
An economical plant regeneration was established for
Caesalpinia bonducella
by culturing immature flower buds and phytoconstituents such as β-Sitosterol (LC3) and methyl (4
E
)-5-(2-[(1
E
)-buta-1,3-dien-1-yl]-4,6-dihydroxyphenyl) pent-4-enoate (SC2) were isolated from its extracts and the isolated phytoconstituents were tested against
Leptospira interrogans.
Objectives:
The aim of the study is to establish the
C. bonducella
through tissue culture technique and to investigate the antileptospiral activity through computational and
in vitro
screening.
Materials
and
Methods:
Morphogenic calli were initiated from 96% of immature flower buds on MS medium supplemented with 2, 4 D (2, 4-Dichlorophenoxyacetic acid) 2.0 mg/l and 6-Benzylaminopurine (BAP) 0.5 mg/l. The calli formed were excised and subcultured on MS medium. Extreme percentage organogenesis (84%) and average shoots per culture were determined on MS medium fortified with 3.0 mg/l BAP and 0.5 mg/l indole-3-butyric acid (IBA). The addition of IBA in ½ MS medium favored the development of recovered shoots. Out of 30 shoots transferred to soil, 27 survived once acclimated. The isolated compounds were selected for
in vitro
and
in silico
screening. The primary pharmacological assay for leptospirosis was carried out by test tube dilution and microdilution technique and the computational screening was done using molecular docking.
Results:
The phytoconstituents obtained from the medicinal plant showed promising results in both
in vitro
and
in silico
antileptospiral activity.
Conclusion:
The LC3 and SC2 compounds isolated from
C. bonducella
were evaluated on Gram-negative bacteria
L. interrogans
. The assay and molecular docking studies revealed the efficacy of phytocomponents of
C. bonducella
as traditional medicine which has an ability to cure bacterial diseases.
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GC-MS/MS-based phytochemical screening of therapeutic potential of
Calligonum polygonoides
L. flower bud against chronic diseases
Mukesh Kumar Berwal, Shravan M Haldhar, Chet Ram, Jagan Singh Gora, Dhurendra Singh, DK Samadia
January-March 2021, 17(5):68-76
DOI
:10.4103/pm.pm_390_20
Background:
Calligonum polygonoides
is an endemic plant species, belongs to Polygonaceae family, native to the "Thar Desert" of India. It is highly tolerant to multiple stresses with dominant biomass and phytochemical producer under extreme niche. It has significant ethnopharmacological applications, but not yet scientifically validated.
Materials and Methods:
The methanolic extract of
C. polygonoides
flower bud was subjected to gas chromatography mass spectroscopy (GC-MS) analysis and antioxidant potential assay was done on different radical scavenging scales. The phytochemicals were identified based on retention time and matching their mass spectra to spectra in NIST 14 library.
Results:
The results revealed the presence of fatty acids, phenolics, terpenoides, flavanoids, alkaloids, tannins, steroids, ketones, esters, and amino acid derivatives, which comprises 93 compounds. Most of the detected compounds have been proved to possess important bio-activities such as anti-microbial, anti-inflammatory, anticancer, anti-diabetic, hepatoprotective, cardiovascular, antioxidant, and antimutagenic. Interestingly, some compounds such as furan-2,5-dimethyl, 2,3-dihydro-2,5-dihydroxy-6 -methyl-4H-pyran-4-one (DDMP), dehydromevalonic lactone, deoxyspergualin, 2-methoxy-4-vinylphenol, benzeneethanol-4-hydroxy-, quinic acid, lauric acid, linolenic acid, and squalene were detected which have proved pharmaceuticals applications against major diseases such as cancer, diabetics, cardiovascular, and some other chronic diseases. Furthermore, the methanolic extract also attributed very high level of antioxidant potential on cupric reducing antioxidant capacity, ferric reducing antioxidant power, 2,2-diphenyl-1-picrylhydrazyl, and phosphomolybdenum assay scales.
Conclusion:
The identified phytochemicals with ample pharmaceutical application explore the worthiness of this endemic plant species. Along with pharmaceutical, it has an immense scope in nutraceutical and functional food industry. These medicinal importance advised for its conservation and artificial regeneration, to sustain the agro-ecological balance of Thar Desert of India.
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125
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Isolation and characterization of flavonoids from fraction of
Blepharis persica
(Burm. f.) O. Kuntze upregulated testosterone biosynthesis
in vitro
using TM3 leydig cells
Nilesh V Gaikar, Manan A Raval, Samir G Patel, Lal L Hingorani
January-March 2021, 17(5):29-37
DOI
:10.4103/pm.pm_406_20
Background:
Seeds of
Blepharis persica
(Burm. f.) O. Kuntze are mentioned in Ayurveda to treat male reproductive functions debilities.
Objectives:
The present study aimed to evaluate the effect of extract and fraction prepared from seeds of
B. persica, in vitro
using TM3 Leydig cells and isolation of constituents from bioactive fraction.
Materials and Methods:
Methanol extract and ethyl acetate fraction prepared that were tested
in vitro
, using TM3 cell line for the testosterone production. Cell viability was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Ethyl acetate fraction of methanol extract was further taken up for detailed chemical investigations, considering biological potential. The fraction was subjected to column chromatography. The column was eluted with a step gradient of the proportion of methanol in chloroform for the separation of compounds. The isolated compounds were characterized using Fourier-transform infrared, nuclear magnetic resonance (
1
H and
13
C), and mass spectroscopy for identification.
Results:
MTT assay performed using TM3 cells revealed that extract and fraction had a half-maximal inhibitory concentration value of more than 300 μg/ml. The methanol extract showed more than half-fold and ethyl acetate fraction of methanolic extract showed a one-fold increase in testosterone at a concentration of 150 μg/ml as compared to nontreated cells. Two constituents isolated from ethyl acetate fraction of methanolic extract were identified as apigenin 7-O-neohesperidoside and naringin using comparative spectral studies.
Conclusion:
The studies suggested that the plant extract and fraction could upregulate testosterone biosynthesis in Leydig cells. Two compounds isolated from the bioactive fraction might serve as a marker for assessment of the quality of plant material.
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880
160
1
Antihyperglycemic activity of
Achyranthes aspera
linn. leaves extract by modulation of β-cell functioning in streptozotocin-induced diabetic rats
Hemant D Une, Trupti C Deshpande
January-March 2021, 17(5):15-20
DOI
:10.4103/pm.pm_296_20
Objective:
The objective of the study is to study the anti-hyperglycemic potential of Ethyl Acetate dissolved fraction of methanolic extract of
Achyranthes aspera
Linn. (EAAA) leaves on STZ induced diabetic rat model.
Materials and Methods:
Safety study of EAAA was carried out with different doses up to 2000 mg/kg. Hyperglycemia was developed in Sprague Dawley male (SD) rats by only one dose of streptozotocin 55 mg/kg in 0.1 M Citrate buffer by intraperitoneal route. Hyperglycemic rats were treated with EAAA 50,100mg/kg and Metformin 120 mg/kg body wt. Biological specifications such as glucose level in blood, insulin level in serum and glucose transporter (GLUT-2) protein expression were measured. In addition, histopathological study of pancreatic tissue was performed to check changes occurred in β-cell complex.
Results:
No mortality was observed during safety study. Weight of treated rats was found to be increased whereas glucose level in blood samples of EAAA treated rats were showed improvement as compare to control groups. The result shows up-regulation of serum insulin levels and small changes in the expression of GLUT-2 after 4-week treatment with the EAAA treatment. The presence of polyphenols in EAAA may have provoked ingestion of glucose by changing the glucose transporter. It has been identified by many researchers that herbal extracts which are rich in polyphenols are responsible for debilitating insulin resistance and thus restrain glucose uptake by expressing GLUT-2 in pancreatic cells of diabetic rats. Histological analysis demonstrated improvement in the structural decay of β-cells of pancreatic tissue.
Conclusion:
The results indicate the thinkable effect of EAAA in STZ-induced diabetic Rats.
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Ethyl acetate fraction of
Anethum graveolens
seeds exerts an antiproliferative effect by inhibiting anti-apoptotic proteins in MCF-7 and PC-3 cells: An
in vitro
and molecular docking study
Furkhan Ahmed Mohammed, Waseem Mohammed Abdul, Md Tabish Rehman, Mohamed F AlAjmi, Fareeduddin Quadri Syed, Muqtadir Baig Mirza, Ayman I Elkady, Anzarul Haque, Muhummadh Khan
January-March 2021, 17(5):105-113
DOI
:10.4103/pm.pm_538_20
Background:
Anethum graveolens
seeds have therapeutic benefits, which may be a potential approach to the treatment of different cancers.
Objectives:
We investigated, the antiproliferative effect of ethyl acetate fraction of dill (EAFD), on MCF-7 and PC-3 cell lines and its two most active components, anethole and carvone by molecular docking analysis.
Materials
and
Methods:
In-vitro
assays, like cell viability assay and measurement of reactive oxygen species, were performed besides performing Giemsa stain and other fluorescent stains; JC-1 dye, dual mixed stain ethidium bromide/acridine orange and 4,6-diamidino-2-phenylindole stain to study morphological characteristics, including molecular docking analysis. EAFD concentrations (0.2, 0.4, 0.6, 0.8, 1.0 mg/ml) were used.
Results:
The EAFD prominently inhibited the proliferation of MCF-7 cells and PC-3 cells by dose-dependent and time-dependent methods. Increased exposure of EAFD to MCF-7 and PC-3 cells increases the level of intracellular oxidative stress. Similarly, EAFD exposure confirms the morphological alternations such as cell shrinkage, membrane disruption, nuclear condensation, and blebbing in phase-contrast microscopy and even fluorescent microscopy stains can lead to mitochondrial membrane degradation, chromatin condensation, nuclear fragmentation. Analyses of docking results suggest that anethole and carvone bind to the hydrophobic patches of Bcl-2 and Bcl-xL through hydrophobic interactions.
Conclusion:
The EAFD may be antiproliferative activity and leading to pro-apoptotic cell death. Molecular docking analysis of Bcl-2 and Bcl-xL anti-apoptotic protein indicated that the antiproliferative activity and pro-apoptotic cell deaths by EAFD are possibly due to inhibition of these proteins.
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Antioxidant and anticancer activities of whole plant extracts of
Lepidagathis pungens
nees:
In vitro
evaluation
Manoharan Dhanalakshmi, Subramaniam Ananda Thangadurai
January-March 2021, 17(5):63-67
DOI
:10.4103/pm.pm_356_20
Background:
Lepidagathis
genus plants were less explored for its biological activity.
Lepidagathis
genus was utilized traditionally for treating of polyuria, fever, urinary calculi, dysentery, dysuria, and uterine disorders.
Objective:
The present study was planned to evaluate antioxidant activities, anticancer activity of
Lepidagathis pungens
Nees., whole plant (
L. pungens
).
Materials
and
Methods:
Antioxidants activity of the ethanol extract of
L. pungens
Nees., determined by total antioxidant assay, diphenyl-2-picryl hydrazyl (DPPH) free radical scavenging assays
,
hydroxyl radical scavenging assays, and nitric oxide scavenging activity assay methods.
Results:
Preliminary phytochemical screening showed that the extract of
L. pungens
possesses flavonoids, alkaloids, tannins, saponins, phenolic, and steroid compounds. In the present investigation, quantitative estimation of phenolic compound and flavonoids carried out by spectrophotometric methods, using aluminum chloride and organic solvent separation method, respectively. The extract showed potent activities in all antioxidant assays and it is also comparable to the standard antioxidants in a dose based manner and remarkable properties to scavenge reactive oxygen species which may have been attributed to the high content of hydrophilic phenolics. The IC
50
values of all parameters were determined and compared with that of the standard. The extract also exhibited potent anticancer activity done by short-term cytotoxicity trypan blue exclusions method with the IC
50
value of 185 μg/ml.
Conclusions:
The results obtained in the present study indicate that ethanolic extract of
L. pungens
is a potent source of natural antioxidants.
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139
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Evaluation of CYP2D, CYP1A2 and distribution of tetrandrine, fangchinoline in the brain, liver, and kidney of wistar rats after short-term exposure to
Cyclea peltata
GI Anuja, VJ Shine, PG Latha, SR Suja, Swapna Susan Abraham, Vinodkumar T G. Nair, S Rajasekharan
January-March 2021, 17(5):77-86
DOI
:10.4103/pm.pm_166_20
Background:
Cyclea peltata
(CP) roots are used in Indian traditional medicine to treat various diseases. However, short-term toxic effects of CP are completely unknown.
Objectives:
The aim of this study is to evaluate short-term toxic effects of CP
,
tissue distribution of bioactive alkaloids tetrandrine, fangchinoline in the brain, liver, kidney, and mRNA expression of CYP2D, CYP1A2 after CP administration.
Materials
and
Methods:
In vitro
toxicity evaluation of CP was carried out using annexin 5/propidium iodide assay with or without CP treatment in L 929 cell line. And in vivo short term toxicity of CP was evaluated in Wistar rats for 28 days. Liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS) based tissue distribution of bioactive molecules was assessed and mRNA expression of CYP1A2, CYP2D, CYP2C6, and CYP2E1 were estimated using qRT- polymerase chain reaction.
Results:
In vitro
toxicity of 70% aqueous ethanol extract of CP 50 μg/mL showed 62.18% cell viability. Oral administration of CP (50, 500 mg/kg) did not cause any clinical, hematological, serum biochemical, and histopathological changes in rats, whereas CP (1000 mg/kg) showed infiltrative changes in the kidney and lungs. Imbalance in oxidative stress and antioxidant defense was reflected as elevated MDA levels in rat liver. LC-Q-TOF-MS studies could detect tissue distribution of bioactive alkaloids tetrandrine, fangchinoline and their decomposed masses in the liver and kidney, whereas tetrandrine and its decomposed molecule (580.3) ([M + H]-43) + could cross blood–brain barrier and were detected in the brain. Evaluation of mRNA expression revealed dose-dependent increase in expression of CYP2D and CYP1A2.
Conclusion:
Oral administration of CP 500 mg/kg for 28 days is safer in rats due to balanced antioxidant defense. Imbalance in antioxidant defense enzymes and toxic metabolites formed through the escalation of CYP2D, CYP1A2 and metabolism of bioactive alkaloids of CP may be the reason for infiltrative changes in kidney, lungs observed after 28 days CP 1000 mg/kg administration.
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Apoptosis-inducing and antiangiogenic activity of partially purified protein from the pericarp of
Zanthoxylum rhetsa in vitro
and
in vivo
Priyanka Dattaraj Naik Parrikar, Balaji Kyathegowdanadoddi Srinivas, Dharmappa Kattepura Krishnappa, Shankar Jayarama
January-March 2021, 17(5):96-104
DOI
:10.4103/pm.pm_520_20
Background:
Biological activities of
Zanthoxylum rhetsa
have been well studied, and its various parts have been reported to possess anticancer activities. The reports on anticancer activities of proteins from the fruits of
Z. rhetsa
are limited.
Objectives:
The study assessed the proapoptotic and antiangiogenic activity of partially purified proteins from the pericarp of
Z. rhetsa
.
Methods:
MCF-7, MDA MB 231, HeLa, and HCT 116 cells treated with partially purified protein fractions of
Z. rhetsa
pericarp were assessed for cytotoxicity by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-diphenyl tetrazolium bromide (MTT) assay. Fraction ZR3 showed high cytotoxicity against the MCF-7 cells; hence, it was chosen for further assays. The proapoptotic activity was scrutinized by Giemsa staining, acridine orange–ethidium bromide staining, and DNA fragmentation assay. The study was supported by wound healing assay and colony formation assay. Cell cycle analysis was performed by fluorescence-activated cell sorter. Ehrlich ascitic carcinoma-bearing Swiss albino mice were used as
in vivo
model. Angiogenesis was studied by peritoneal angiogenesis in mice and chorioallantoic membrane assay in fertilized eggs.
Results:
MTT assay showed the inhibition of MCF-7 cells (IC
50
= 21.5 μg/mL) by ZR3 fraction. Reduction in proliferation and failure to produce large cell colonies were observed. Proapoptotic activity was evident from the DNA fragmentation and staining methods. ZR3 blocked the cells in the G2/M phase of the cell cycle.
In vivo
studies suggested the antiproliferative and proapoptotic activities of ZR3. ZR3 exhibited antiangiogenic properties
in vivo
.
Conclusion:
This study confirmed the role of
Z. rhetsa
partially purified proteins as a potential proapoptotic and antiangiogenic agent against the MCF-7 cell line.
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