Antiplasmodial Potential of Sesuvium portulacastrum(L.) L

Abstract

This is an Open Access Journal / article distributed under the terms of the Creative Commons Attribution License (CC BY-NC-ND 3.0) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. All rights reserved. Halophytic flora is crucial for preserving ecological stability and safeguarding coastal ecosystems. Numerous secondary metabolites found in halophytes play an essential role in biological processes. Sesuvium portulacastrum(L.) L. is a herbaceous, eternal, dichotomous, culinary halophyte that grows quickly. It contains active phytochemical constituents such as alkaloids, saponins, tannins and terpenoids which are responsible for many biological activities. The leaves of this plant have been subjected to hot continuous extraction using the soxhlet method. From polar to non-polar, nine different solvent types were selected for crude extraction. The Plasmodium falciparum (3D7) strain is cultivated using the Trager and Jensen technique (1976). A polar solvent, ethanol, possess moderate activity with an inhibition percentage of 46.30±1.20µg/mlcompared with the other extracts among the nine solvents. From this study it is observed that the plant S. portulacastrum showed some moderate activity against P. falciparum. In higher concentration of the plant extract the inhibition percentage also increases. ABSTRACT RESEARCH ARTICLE

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Indian Journal of Natural Sciences www.tnsroindia.org.in ©IJONS
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57856
Yashoda
Antiplasmodial Potential of Sesuvium portulacastrum(L.) L.
Deebika Kumar1, D. Kumarasamy2* , Ritu Gill3, Archana Sharma4, Kathiravan Mani1 and P. Munnaji1
1Ph.D Scholar, Department of Botany, Annamalai University, Chidambaram- 608002, Tamil Nadu, India.
2Professor, Department of Botany, Annamalai University, Chidambaram- 608002, Tamil Nadu, India.
3Assistant Professor, Centre for Biotechnology, Maharishi Dayanand University, Rohtak -124001,
Haryana ,India.
4Ph.D Scholar, Centre for Biotechnology, Maharishi Dayanand University, Rohtak-124001, Haryana ,India
Received: 06 May 2023 Revised: 07 June 2023 Accepted: 10 July 2023
*Address for Correspondence
D. Kumarasamy
Professor,
Department of Botany,
Annamalai University,
Chidambaram- 608002,
Tamil Nadu, India.
E.Mail: drdkumarasamy@gmail.com
This is an Open Access Journal / article distributed under the terms of the Creative Commons Attribution License
(CC BY-NC-ND 3.0) which permits unrestricted use, distribution, and reproduction in any medium, provided the
original work is properly cited. All rights reserved.
Halophytic flora is crucial for preserving ecological stability and safeguarding coastal ecosystems.
Numerous secondary metabolites found in halophytes play an essential role in biological processes.
Sesuvium portulacastrum(L.) L. is a herbaceous, eternal, dichotomous, culinary halophyte that grows
quickly. It contains active phytochemical constituents such as alkaloids, saponins, tannins and terpenoids
which are responsible for many biological activities. The leaves of this plant have been subjected to hot
continuous extraction using the soxhlet method. From polar to non-polar, nine different solvent types
were selected for crude extraction. The Plasmodium falciparum (3D7) strain is cultivated using the Trager
and Jensen technique (1976). A polar solvent, ethanol, possess moderate activity with an inhibition
percentage of 46.30±1.20µg/mlcompared with the other extracts among the nine solvents. From this study
it is observed that the plant S. portulacastrum showed some moderate activity against P. falciparum. In
higher concentration of the plant extract the inhibition percentage also increases.
Keywords: S. portulacastrum, Plasmodium falciparum, saponins, metabolites, solvent.
ABSTRACT
RESEARCH ARTICLE

Indian Journal of Natural Sciences www.tnsroindia.org.in ©IJONS
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INTRODUCTION
Halophytes are plants that thrive in a wide range of saline environments, including inland deserts, salt flats, steppes,
coastal dunes, salt marshes, and mudflats (1). In addition to their salt tolerance thresholds, these plants exhibit a high
degree of physiological adaptability depending on their origin climate zone (2). Due to their high polyphenol
content, halophytes exhibit remarkable physiological flexibility (3, 4). Bioactive antiviral, antibacterial, and antifungal
compounds are found in mangroves and their associates. Steroids, triterpenes, saponins, flavonoids, alkaloids, and
tannins are abundant in them (5, 6, 7, 8). Aizoaceae family member Sesuvium portulacastrum (L.) is a significant
halophyte in the group of "salt accumulator" plants, which amass high salt concentrations in their cells and tissues
and avoid salt poisoning by generating succulence (9). It is considered a potential candidate for environmental
conservation due to its capacity to endure under challenging environmental circumstances (10). Conventional
medicine use this plant as a treatment for scurvy, renal issues, and fever (11). Five different single-cell,
eukaryotic Plasmodium parasite species that are spread by the bite of Anopheles mosquitoes—primarily Plasmodium
falciparum and Plasmodium vivax—cause malaria in humans. Worldwide, there were an anticipated 247 million
malaria cases in 2021 in 84 malaria-endemic countries (including the territory of French Guiana), up from 245 million
in 2020, with the majority of this rise coming from countries in the WHO African Area (12).Development of efficient
antimalarial medications is hampered by the evolution of parasite strains that are resistant to treatments (13).This
research revealed that there is a knowledge gap in this study since there is a paucity of data that is supported by
evidence because this plant's ability to treat malaria has not been previously reported.
MATERIALS AND METHODS
Collection of Plant Material
The Plant materials (Leaves) were collected from Kodiyamplayam (Latitude 11.3822° N and Longitude 79.8136° E),
which is a portion of the Pichavaram mangrove cover, South East coast of India, Tamilnadu. The gathered plant
material was taxonomically recognised and a voucher specimen was kept in the Herbarium of the Botany
department at Annamalai University (Voucher No. 69).The collected plant materials are washed with tap water and
then with distilled water to remove adhering salts and other associated animals. The plant materials are subjected to
shade-dried for 2-3 weeks.
Extract preparation
Shade-dried leaves are made into a coarse powder (100g)using a mechanical blender and it is further subjected to hot
continuous extraction by soxhlet apparatus (according to the boiling point of the solvent). The solvents chosen for the
extraction process include n-hexane, Diethyl ether, Ethyl acetate, chloroform, dichloromethane, acetone, n-butanol,
ethanol, and methanol (300ml).
In vitro culture of Plasmodium falciparum
CQ-sensitive strain 3D7 of P. falciparum was used for in vitro blood stage culture to test the antimalarial efficacy of
different plant extracts. The strain 3D7 was obtained from Jawaharlal Nehru University, New Delhi. Plasmodium
falciparum culture was maintained according to the candle jar method described by Trager and Jensen (1976), with
minor modifications. For antimalarial activity evaluation, highly synchronous ring stage P. falciparum was used in
each assay. The culture was maintained in fresh O+ve human erythrocytes suspended at 4% hematocrit in RPedMI
1640 (Himedia) containing 0.2% sodium bicarbonate, 0.5% albumax, 2 % glucose, 45 μg/L hypoxanthine, and 50 μg/L
gentamicin(Himedia) and incubated at 37°C under a gas mixture of 5% O2, 5% CO2, and 90% N2. Every day, infected
erythrocytes were transferred into a fresh complete medium to propagate the culture.
Deebika Kumar
et al.,

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57858
Antiplasmodial assay
Filter sterilized leaf extracts (200, 100, 50, 25, 12.5, 6.25 µg/mL) was incorporated in 96 well tissue culture plates
containing 100 µL of P. falciparum culture with fresh red blood cells diluted to 2% hematocrit. Negative control was
maintained with fresh red blood cells and 2% parasitized P. falciparum diluted to 2% hematocrit, and positive control
was maintained with parasitized blood cells culture treated with chloroquine (14). Parasitaemia was evaluated after
48 h by Giemsa stain and the average percentage suppression of parasitaemia was calculated by the following
formula:
Average % suppression of Parasitemia = .  
 .  ×100
Chemical injury to erythrocytes
To assess the chemical injury of the leaf extract on erythrocytes, 100 µL of erythrocytes were incubated with 200
µg/mL of the extract. The conditions of the experiment were maintained as in the case of the antiplasmodial assay.
After 48 h of incubation, thin blood smears were stained with Giemsa stain and observed for morphological changes
under high-power light microscope(15). The morphological findings were compared with those in erythrocytes that
were uninfected and not exposed to the extract.
SYBR green assay: After 48 h of incubation, 100 µl of SYBR green dye was added to each well. 0.2 µl of SYBR green
dye per ml of PBS lysis buffer and this experiment is carried out under a light-sensitive area (16). The reading was
recorded in (excitation at 480 nm, and emission at 520 nm)
Antiplasmodial activity calculation and analysis
The antiplasmodial activity of mangrove leaf extracts was expressed by the inhibitory concentration 50 (IC50),
representing the concentration of drug that induced a 50% parasitaemia decrease compared to the positive control
culture referred to as 100% parasitaemia (17).
Statistical analysis
The IC50 values were calculated (concentration of extract in X axis and percentage of inhibition in Y axis) using Graph
Pad Prism software with the linear regression equation.
RESULTS AND DISCUSSION
The antiplasmodial potential of Sesuvium portulacastrum is listed in Table No. 1. The Nine type of solvents were taken
and screened against Plasmodium falciparum. The concentration of the plant crude extracts are 200 µg/ml, 100 µg/ml,
50 µg/ml, 25 µg/ml, 12.5 µg/ml, 6.25 µg/ml. Among all the extracts ethanol shows better activity with the inhibition
percentage of 46.30±1.20 µg/ml towards chloroquinine sensitive 3D7 strain of P. falciparum. The positive control
chloroquine showed 50 % inhibition with IC50 value 41.89±1.89 µg/ml. Except positive control, all the other plant
extracts showed IC50 value greater than 200 µg/ml (IC50 ≥ 200).
Traditional practitioners employ the entire plant for numerous diseases due to the presence of many bioactive
components(18). This halophyte has been utilised for centuries as conventional healthcare to treat conditions
including toothaches, leprosy, eye inflammation, dermatitis, haematuria, and conjunctivitis(19).Antibacterial,
antifungal, and antioxidant properties were present in the essential oil extract of this plant(20).It contains phenol,
which is an essential source of antioxidants(21). The MDA-MB-231, IMR-32, and HCT-116 cell lines were susceptible
to the anticancer effects of S. portulacastrum diethyl ether extracts in a dose-dependent manner(22).The chemical
components pyrrole derivatives, butanoic acid, ascorbic acid, octadecanoic acid, and hentriacontane, which act as
antioxidants, antimicrobials, antiulcerogenic agents, and anticancer agents, were detected in the GC-MS analysis of
the methanolic extracts of S. portulacastrum (23).
Deebika Kumar
et al.,

Indian Journal of Natural Sciences www.tnsroindia.org.in ©IJONS
Vol.14 / Issue 79 / Aug / 2023 International Bimonthly (Print) – Open Access ISSN: 0976 – 0997
57859
CONCLUSION
The current investigation concluded that ethanolic extracts of Sesuvium portulacastrum have modest antiplasmodial
action against the 3D7 strain of Plasmodium falciparum. The antiplasmodial action of Sesuvium portulacastrum has
never been reported before. In higher concentrations of plant crude extracts, the inhibition percentage also increases
with lower IC50 value.
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Table No. 1 Antiplasmodial Potential of Sesuvium portulacastrum
6.25
µ
g/ml
12.50 µ
g/ml
25 µ
g/ml
50 µ
g/ml
100 µ
g/ml
200 µ
g/ml
Hexane
21.10±1.82
24.68±2.59
25.47±2.94
27.62±2.78
30.43±2.56
37.65±4.33
Diethyl ether
7.22±0.55
11.28±2.90
12.71±2.33
14.05±2.06
17.34±2.25
19.41±2.54
Ethyl acetate
21.33±0.59
25.46±1.09
26.16±1.00
27.51±1.07
29.29±1.61
35.97±0.81
Chloroform
11.29±2.22
14.41±1.90
17.52±2.28
19.97±1.91
22.30±1.61
25.68±0.82
Dichlorometh
ane
18.32±1.84
21.42±1.97
23.30±0.90
24.14±1.12
25.07±0.73
25.26±0.67
Acetone
13.59±2.19
18.38±5.67
22.65±3.25
23.50±3.59
25.50±1.48
30.19±1.84
Ethanol
25.42±1.11
29.81±0.98
35.97±1.72
38.74±0.76
44.20±0.94
46.30±1.20
Methanol
13.71±0.57
16.97±2.14
23.07±1.68
28.10±2.56
36.73±0.63
41.24±1.62
Butanol
13.91±1.92
18.71±1.80
21.19±2.22
21.43±2.11
27.17±6.94
31.91±3.37
Deebika Kumar
et al.,