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WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES
Kabbash World Journal of Pharmacy and Pharmaceutical Sciences

SJIF Impact Factor 6.041
Volume 5, Issue 6, 84-100 Research Article ISSN 2278 – 4357
MACROSCOPIC AND MICROSCOPIC CHARACHTERIZATION OF

ATRIPLEX HALIMUS L. GROWING IN EGYPT AND IN VITRO
EVALUATION OF ITS CYTOTOXIC ACTIVITY
Amal M. Kabbash*
Department of Pharmacognosy, Faculty of Pharmacy, Tanta University, Tanta (31527),

Egypt.
ABSTRACT
Article Received on
26 March 2016, Herbal medicines have been commonly used and remain so instead of
Revised on 17 April 2016,
Accepted on 08 May 2016
synthetic drugs because of their possible fewer side effects. Atriplex
DOI: 10.20959/wjpps20166-6854 halimus L. (Amaranthaceae), denominated saltbush, is a promising
medicinal plant growing in Egypt. The aim of this study is to address
*Corresponding Author the botanical identification of this valuable plant and evaluate its
Amal M. Kabbash cytotoxic activity. Transverse sections and macerated tissues were
Department of examined. From present evaluation, it is evident that certain characters
Pharmacognosy, Faculty
such as hair, stomata in epidermal layer, calcium oxalate, vascular
of Pharmacy, Tanta
bundles and xylem vessels can provide useful information for correct
University, Tanta (31527),
Egypt. identification of the plant. In addition, quantitative measurement of
different cells dimensions is presented as an evidence for correct
identification using Orion software version 6. Cytotoxicity (SRB assay) of the plant total
extract and different fractions was assessed using cells from human hepatocellular (HepG2),
colon (HCT), intestinal (CACO), larynx (HEP2), cervical (HELA), lung (A549), breast
(MCF-7), and prostate (PC3) carcinoma and doxorubicin was used as a standard. The results
showed that the total methanol-methylene chloride (1:1) extract inhibited the survival of the
different tested cell lines by 67±0.76, 75±1.7, 62.5±0.9, 73.5±1.14, 73.0±1.3, 74.5±1.76,
80.5±1.5, and 78.0±0.67 %, respectively. Furthermore, the cytotoxic activities of different
fractions obtained after MCI-gel CHP20P column fractionation were evaluated. Fractions
eluted with water, and H2O-MeOH (40:60) showed the highest activities against prostate and
breast carcinoma cell lines with IC50 of 43.7±0.56, and 25±0.87 µg/mL, respectively. It was
concluded that Atriplex halimus L. could be correctly identified using some characteristic
macroscopic and microscopic features together with quantitative determination of different
www.wjpps.com Vol 5, Issue 6, 2016. 84

cells dimensions. The total extract and different fractions of A. halimus L. possess a dose
dependent cytotoxic activity with specific selectivity against MCF-7 and PC3 carcinoma
cells.
KEYWORDS: Atriplex halimus L., Macroscopic, Microscopic, Cytotoxicity, MCF-7 and

PC3 carcinoma cells.
INTRODUCTION
Plants play a major role for the discovery of new therapeutic agents and have received much
attention as sources of biologically active substances.[1] Atriplex halimus L. is one of those
plants used by tribes to treat diabetes and heart conditions. Atriplex has always been a
popular folk remedy. In the Arab word, A. halimus L. was used to treat chest ailments, as a
laxative, to cure stomach pains, for intestinal worms, and to regulate gall bladder
excretions.[1,2] The Bedouin tribes of the Negev use the plant today for treating muscular pain.
Other uses the Bedouin have found for A. halimus L. were to feed sheep and goats, to treat
intestinal diseases in animals and to flavor cooking.[3] A. halimus L. is also a source of
vitamins A, C and D[4], tannins, flavonoids, saponins, alkaloids and resins, and it contains up
to 10% sodium chloride.[5] Atriplex species are among the few salt-tolerant plants that have
many agricultural values in extremely arid and saline areas because of their unusually high
protein content and exceptional salt tolerance.[6] A halimus L. is a xero-halophyte species
belonging to Chenopodiaceae widely distributed in non-saline as well as saline areas, in sub-
humid to arid regions of South Europe, East Mediterranean and North Africa, including the
Sahara in Algeria.[1,7] This plant is often cultivated in forage because tolerating severe
conditions of drought, and it can grow up in very alkaline and saline soils. [3] The reported
data in the current literature describing the morphological and anatomical features of A.
halimus L. growing in Egypt is scarce. These data are required for the identification
procedures to meet the quality standard. Even though some sophisticated chemical and
molecular method are available for identification of the plant material, morphological
and anatomical identifications are simplest among the qualitative methods. Hence it becomes
a useful tool to determine characteristics of the plant material to avoid falsification and
adulteration of the drugs. Given these considerations, this study was taken up to investigate
the morphological and anatomical characteristics of A. halimus L. grown in Egypt and to
evaluate its cytotoxic activity which, to our knowledge, is the first to demonstrate the
cytotoxic activity of A. halimus L. using the different tested cell lines.

MATERIALS AND METHODS

Collection and identification of plant materials
The aerial and underground parts of Atriplex halimus L. (Amaranthaceae) were collected
from International road (Balteem area) in February, 2014 and identified by Prof. Kamal
Shaltoot, professor of taxonomy, Department of Botany, Faculty of Sciences, Tanta
University-Egypt. A voucher specimen was deposited at the international herbarium of
Faculty of Sciences, Tanta University, Egypt.
Chemicals
Formalin, acetic acid, ethanol, xylol, safranin, dimethyl sulfoxide (DMSO), trichloroacetic
acid, sulforhodamine B (SRB), ethylenediaminetetraacetic acid buffer (EDTA) and acetic
acid were purchased from Sigma- Aldich Co. Potassium hydroxide used was of analytical
grade. MCI-gel CHP20P was purchased from Mitsubishi Chemical Co., Tokyo, Japan. All
chemical and solvents used in this study were of analytical grades. Liver carcinoma cell line
(HepG2), colon carcinoma cell line (HCT), intestinal carcinoma cell line (CACO), larynx
carcinoma cell line (HEP2), cervical carcinoma cell line (HELA), lung carcinoma cell line
(A549), breast carcinoma cell line (MCF-7), and prostate carcinoma cell line (PC3) were
obtained from the American Type Culture Collection (ATTC).
Microscopic study
Sections of leaf, old, middle, and young stem, and root, together with macerated tissues of the
plant were prepared for microscopic study. The staining was done using standard laboratory
methods.[8,9] Fresh plant material was fixed in formalin-acetic acid-70% ethanol (5:5:90,
V/V).[10] The leaf, old, middle and young stem, and root were sectioned using rotary
microtome to the thickness of 10 to 30 µm. All sections were stained in safranin (1% solution
in 50% ethanol) for 15 minutes, washed in ethanol 50, 70 and 100%, respectively for 5
minutes, and then immersed in xylol for 5 minutes. The slides were dried at 50 oC for 72
hours. Samples of the tissues from different organs were macerated with boiling KOH for 20
minutes and used for microscopical identification. Microphotographs were obtained using
light microscope Olympus CX31 with camera – Japan, using different magnification powers.
Quantitative measurement of different cells dimensions is presented using Orion software
version 6 (data are presented in table 1).

Determination of Cytotoxicity
The total plant extract was prepared by maceration of the air-dried aerial parts of powdered
plant with absolute methanol-CH2Cl2 (1:1) overnight, and the extraction was repeated till
exhaustion. Fractionation of Atriplex halimus L. was carried out using 200 g of air-dried
aerial parts of powdered plant and extraction with MeOH by sonication for 30 min. at room
temperature till exhaustion. The extract was concentrated under reduced pressure to afford a
residue (3.5 g). The residue was partitioned between n-hexane and 80 % MeOH, after which,
the 80 % MeOH layer was concentrated to yield a residue (1.7 g), which was loaded onto
MCI-gel CHP20P column (50×300 mm) and eluted sequentially with H2O; H2O-MeOH
(80:20); H2O-MeOH ( 60:40); H2O-MeOH (40:60); H2O-MeOH (20:80); 100% MeOH, and
acetone to afford seven fractions (Fr.1–7), respectively. For screening of the cytotoxic
activity, the sample stock solution was prepared at concentration of 1mg/mL using
dimethylsulfoxide (DMSO) as solvent. Cytotoxic activity was performed in collaboration
with Egyptian National Cancer Institute.
The procedures were carried out according to the method reported by Skehan et al. [11] The
methanolic-methylene chloride (1:1) extract of Atriplex halimus L. and the different fractions
obtained after MCI-gel CHP20P column fractionation were tested using a single dose
experiment at a dose of 100 µg/mL in DMSO. A multi dose experiment was conducted to
evaluate the activities of both total extract and different fractions against the most susceptible
cell lines (MCF-7 and PC3). The % inhibition against the different tested cell lines were
determined and the results are presented as mean % inhibition ±SD (figure 8 and tables 2, 3,
and 4).
RESULTS AND DISCUSSION

Macroscopic study
Atriplex halimus L. is a highly polymorphic species that is widespread, perennial,
monoecious and polygamous.[12] The evidence for its morphological and anatomical
characterizations in literature is scarce. Morphologically (figure 1), A. halimus L. is an erect,
perennial shrub, 0.5-1.7 m. Leaves are broadly rhombic, entire, glabrous, alternate
(phyllotaxis =1/5) and petiolated. The leaf measures 2.2-5.0 cm × 3-4.2 cm. The leaves are
glaucous, and leaf petiole is cylindrical pale green and measures 1-1.4 cm in length and 0.2 -
0.3 cm in diameter. The stem is smooth and herbaceous. Stem is nearly cylindrical in shape,
greenish to reddish green in color, and solid. The root is cylindrical in shape, with rough

surface, and brown in color, it measures about 50-70 cm in length and 3-5 mm in diameter.
The main root is erect, extends below the soil, it is more tough and harder than the smaller
branches, which extend laterally below the soil. Most of the morphological characteristics of
this plant are in accordance to that reported in the Egyptian flora.[13]
Microscopic study
The transvers section of the leaf (figure 2) shows the midrib being prominent on both lower
and upper surface. The epidermis consists of one row of subrectangular cells, covered with
numerous vesiculated bladders (hairs) on both surfaces similar to those described by Black on
two species of Atriplex from Australia[14], and by Brian and Catilin on Chenopodium album
L.[15] The bladders may be implicated in the tolerance of the Atriplex species to saline
areas.[16] Hypodermis is situated directly below the upper epidermis in the lamina area; it
consists of one row of square to subrectangular tangentially elongated cells, which are
discontinued in the midrib region. The leaves had Kranz-type leaf anatomy. A definition of
this type of leaf anatomy in Atriplex was given by Frankton and Bassett.[17] Each vascular
bundle was surrounded by a layer of bundle sheath cells and radially-arranged palisade cells
forming concentric centers around the vascular bundle (wreathlike arrangement). The bundle-
sheath cells were well developed and occupied a substantial portion of the leaf (figure 2-b).
Kranz anatomy is associated with the four-carbon (C4) photosynthetic pathway, which is a
metabolism of high photosynthetic efficiency.[18] In the midrib region, the upper epidermis is
followed by rounded cellulosic collenchyma cells forming 2-3 rows, which followed by
rounded parenchyma cells, forming (4-5) rows. The large parenchyma in midrib region
contains large cluster crystals of calcium oxalate (figure 2-c and e). The vascular system
composed of four collateral vascular bundles (figure 2: a, d). The endodermis surrounds the
stele; it consists of one single layer of tangentially elongated cells. Quantitative measurement
of different types of cell is presented (table 1).
A plant is an opened biological system, that due to its subsystems, i. e. its histo-anatomical
features, might provide data on certain physico-chemical particularities on the environment.
Successive cambia, met within halophytes as the Chenopodiaceae, is considered by most
researchers a structural anomaly it affects axial vegetative organs, which are the root and the
stem.[19] Transverse section of old stem showed that the outline of the stem was almost
circular (figure 3-a). The section showed the structures as follows: The epidermis was an
outermost layer of relatively thin barrel to rectangular cells. The cortex was multi-layered and

differentiated into collenchyma and parenchyma. A few layered collenchyma hypodermis (1-
3) layered deep follows epidermis. Parenchyma followed collenchyma hypodermis and was
(3-4) layered cells. The cells were spherical to oval. Large cluster crystals of calcium oxalate
were scattered inside the parenchyma cells. The cortex is delimited internally by a band of
lignified sclerenchyma (Figure 3: a, b, e and f). The successive cambia resulting a large
number of xylemic-phloemic concentric rings. The vascular strands are separated by
cellulosic parenchyma cells. The secondary xylem of the plant is a solid cylinder and exhibits
what is known as anomalous secondary growth.[19,20] These cylinders included phloem or
interxylary phloem to the inside of the ring of collateral vascular bundles. This region
contains parenchyma cells and large cluster crystals of calcium oxalate. Pith is
relatively large and made up of paranchymatous tissue. Most of the anatomical features of the
stem are consistent with that reported for the development of the successive cambia in
Atriplex halimus L.[20] Quantitative measurements of different types of cell are presented
(table 1).
Transverse section of the middle stem is circular, showing limited, and less prominent
patches of collenchyma tissue (figure 4). The cortex is delimited internally by a weak, none
defined band of lignified sclerenchyma, which is differ than that of the defined, lignified one
of old stem. The successive cambia resulting in a limited number of xylemic-phloemic
concentric rings than that in case of old stem. The vascular strands are separated by cellulosic
parenchyma cells. Cluster crystals of calcium oxalate were scattered inside the parenchyma
cells. Pith is relatively larger than in case of old stem. There was significant decrease in all
cells dimensions comparing to old stem. Quantitative measurements of different types of cell
are presented (table 1).
Transverse section of the young stem is circular, showing 6-8 ridges with patches of
collenchyma tissue (figure 5: a, c and d). The epidermis is covered with bladder cells (salt
gland) similar to those of the leaves. The cortex is differentiated into collenchyma and
parenchyma. The outer region of collenchyma is clearly developed at the ridges. The cortex is
delimited internally by a sheath contains cluster crystals of calcium oxalate (replaced by a
band of lignified sclerenchyma in case of old stem). The primary collateral vascular bundles
surround wide pith which contains cluster crystals of calcium oxalate. There is none defined
secondary growth because accessory cambia have not yet develop.[20] There was significant

decrease in all cells dimensions comparing to old and middle stem. Quantitative
measurements different types of cell are presented (table 1).
In the Chenopodiaceae, secondary thickening deviates from that typical of most dicotyledons
and is referred to as anomalous secondary thickening.[21] This type of thickening was
regarded as being derived from numerous successive cambia.[20] Transverse section of the
root (figure 6) showed anomalous secondary thickening with absence of pith. The exodermis
was an outermost layer of barrel to rectangular cells. The cells were covered with cuticle and
followed by multi-layered parenchyma (3-4) layered deep. The root has a number of (6-7)
concentric xylem rings separated by an equal number of phloemic rings. The xylemic and
phloemic rings are interrupted or in touch with the other. Xylemic parenchyma has cellulosic
walls and the libriform is represented by moderate thick and lignified-wall fibers. The
xylemic fibers that dominate the xylemic structure have a strong, thickened architecture and
lignified. The center of the root is made mostly of a compact, massive, and highly lignified
xylem as shown in figure 6.
Section of macerated whole plant tissues was prepared using KOH and examined using light
microscope to reveal the different characteristic elements of the plant. The obtained
microphotographs (figure 7) showed the presence of cluster crystals of calcium oxalate
arranged in crystal layers and spiral xylem vessels. The stomata were found to be anomocytic
type.
Cytotoxic study
Atriplex halimus L. is well known and extensively used to treat diabetes especially in the
Middle East.[22] The cytotoxic effect of 50% ethanol/water extract of A. halimus L. was
previously measured with MTT assay and the LDH leakage assay using liver hepatocellular
carcinoma cell line (HepG2) and cells from the rat L6 muscle cell line. This previous study
revealed that A. halimus L exhibited cytototoxic effect at concentrations higher than 500
𝜇g/mL.[23] In the present study, methanol-methylene chloride (1:1) extract of A. halimus L.
was used instead of the methanolic or ethanolic extract to avoid the excessive salt present in
the plant. The cytotoxic effect of methanol-methylene chloride (1:1) extract of Atriplex
halimus L. was evaluated using cells from the human hepatocellular carcinoma (HepG2),
colon carcinoma (HCT), intestinal carcinoma (CACO), larynx carcinoma (HEP2), cervical
carcinoma (HELA), lung carcinoma (A549), breast carcinoma (MCF-7), and prostate
carcinoma (PC3) human cell lines. SRB assay was used for evaluation of the cytotoxic

activity of the total extract and different fractions from Atriplex halimus L. This assay is
widely used in in vitro toxicology and for the detection of cytotoxic and other negative
effects on cell viability following exposure to test materials. The results obtained from this
study (tables 2-4, and figure 8) indicate that Atriplex halimus L is cytotoxic at concentration
100 𝜇g/mL. There was a reduction in cells viability of all the tested cell lines (> 60%).
However, Atriplex halimus L. methanol-methylene chloride (1:1) extract showed the highest
cytotoxicity to breast carcinoma (MCF-7) cells (80.5±1.5) and prostate (PC3) carcinoma cells
(78.0±0.67) at a dose of 100𝜇g/mL (figure 8). A multi dose experiment was carried out to
evaluate the cytotoxicity of the extract against breast (MCF-7) and prostate carcinoma cells
(PC3) using doxorubicin as a standard. The results showed that the total extract exhibited a
moderate activities (47.57±1.2 and 43.0±1.8 %) against breast and prostate carcinoma cell
lines, respectively at a dose of 50 µg/mL with IC50 >50 µg/mL. This results indicate the dose
dependency of the cytotoxic activity of the extract (compared to 80.5 and 78.0% inhibition at
a dose of 100 µg/mL). Furthermore, the cytotoxic activities of different fractions obtained
after MCI-gel CHP20P column fractionation were evaluated for the different fractions (1-7).
The fractions presented variable activities against both breast and prostate carcinoma.
Fractions 1 (eluted with water) and 4 (eluted with H2O-MeOH, 40:60) showed the highest
activities compared to other fractions obtained from MCI-gel CHP20P column. The multi
dose experiment cytotoxic evaluation of fractions 1 and 4 revealed that both fractions had
cytotoxic activities against breast and prostate carcinoma cells with IC50 of 25±0.87 and 43.7
±0.56 µg/mL, respectively. The obtained results revealed that the method of extraction and
fractionation significantly affect the cytotoxic activity of A. halimus L., and the activity was
dose-dependent. Comparing to the cytotoxic effect of doxorubicin, which is pure compound
and used as positive control in this study, a high dose of the plant extract/fractions is required
to obtain a maximum activity because they contain a mixture of different phytochemicals and
some impurities which may not participate in the cytotoxic activity.
Figure 1: Morphological characteristics of Atriplex halimus L.

Figure 2: Transverse section of Atriplex halimus L. leaf.

a: T.S of the whole leaf showing the structural arrangement of different layers (X100); b: T.S
in lamina showing Kranz-type leaf anatomy (X400); c: Upper epidermis and part of
mesophyll with parenchyma cells containing clusters of CaOx (X400); d: mesophyll showing
vascular bundles (X200); e: part of midrib region (X200); f: vascular bundles region (X400).

Figure 3: Transverse section of old stem.

a: T.S of old stem showing the structural arrangement of different layers and merging of
rings of vascular strands (X40); b: structural arrangement of different layers (X100); c:
cortical region and vascular bundles (X200); d: T.S of old stem showing vascular bundles and
phloem (X200); e: cortical layers, endodermis, and phloem (X400); f: parenchyma cells of
cortical layer containing cluster crystals of calcium oxalate (X400).

Figure 4: Transverse section of middle stem

a: T.S of middle stem showing the structural arrangement of different layers (X200); b: T.S
of middle stem showing the cortical region (X200); c: T.S of middle stem showing the
xylem and phloem (X400) ; d: T.S of middle stem showing the xylem vessels (X400).
Figure 5: Transverse section of young and middle stem

a: T.S of young stem showing the structural arrangement of different layers (X40); b: T.S of
middle stem showing the structural arrangement of different layers (X40); c: T.S of young
stem showing the structural arrangement of different layers (X100) ; d: T.S of cortical region
of young stem (X400).

Figure 6: Transverse section of root

a: T.S. of the root showing the structural arrangement of different layers (X40) ; b and c: T.S
of the root showing typical secondary thickening with absence of pith (X100, 200
respectively); d: cortical region and typical secondary thickening; e: cortical layers and part
of typical secondary thickening (X400); f: exodermal cells, cortical layers and part of xylem
vessels (X400).

Figure 7: Microscopic characterization of A. halimus L. using macerated tissues

a: anomocytic stomata (X400); b: spiral xylem vessel (X100), c: cluster crystals of calcium
oxalate (X200) ; d: calcium oxalate arranged in crystal layer (X100).
Figure (8): Cytotoxic activity of Atriplex halimus L. against different cell lines

Table 1: Measurements of different cell types of leaf, stem, and root of Atriplex halimus
L.
Breadth (µm) Length (µm)
Plant Part Cell type Min Max Min Max
±SE ±SE ±SE ±SE
Leaf Bladder cell 35±1.2 41±0.67 52±0.98 62±1.0
Epidermal cell 17±0.89 42±0.89 32±0.76 44±0.66
Parenchyma cell of hypoderm 26±0.096 46±1.0
Collenchyma cell 11±1.1 20±0.94
Palisade cell 12±1.3 21±0.92 25±0.78 37±0.87
Bundle sheath cell 11±0.7 21±1.4 24±0.99 38±0.55
Parenchyma cell of the midrib 35±0.87 51±1.1
Phloem cell 5±0.79 7±0.89
Xylem vessel 10±1.1 26±0.67
Wood parenchyma cell 24±1.0 360.77
Old stem Epidermal cell 16±0.49 20±1.0 56±1.1 70±0.78
Parenchyma cell (cortex) 25±0.76 40±0.88 64±0.56 68±0.88
Sclerenchyma cell 17±1.2 19±1.0 70±0.97 72±1.3
Endodermal cell 28±0.69 32±1.2
Parenchyma cell (pericycle) 19±68 35±0.78
Phloem 14±1.6 18±0.98
Sclerenchymatic fiber 17±2.0 18±1.0 70±1.4 72±0.66
Parenchyma cell (pith) 54±1.02 69±0.98
Mid. stem Epidermal cell 13±0.9 15±0.68 15±0.46 30±0.57
Parenchyma cell (cortex) 18±0.76 22±0.88 54±0.28 60±0.68
Sclerenchyma cell 12±1.1 17±0.97 15±0.78 30±0.66
Endodermal cell 16±0.98 26±1.1
Parenchyma cell (pericycle) 12±0.76 30±0.56
Phloem 9±1.2 15±0.55
Young stem Epidermal cell 12±0.68 14±1.0
Parenchyma cell (cortex) 21±0.77 34±0.35
Root Exodermal cell 9±0.89 12±0.57 44±0.77 56±1.0
Parenchyma 19±1.0 25±0.57
Phloem parenchyma 40±0.94 42±0.67
Phloem 15±0.87 19±1.0

Table 2: Multi Dose cytotoxic evaluation of total extract of A. halimus L. on MCF-7 and
PC3 cells
Non Surviving Fraction* (%)±SD
Conc.
MCF-7 cells PC3 cells MCF-7 cells PC3 cells
(µg/mL)
(Total extract) (Doxorubicin)
100 75.90±1.0 77.56±1.3 88±0.92 78.40±1.3
50.0 47.57±1.2 43.0±1.8 79.87±2.6 61.1±1.8
25.0 36.90±0.42 32.21±1.1 71.55±0.99 56.5±1.9
12.5 19.30±0.52 19.50±0.67 70.83±0.92 54.4±0.82
5.0 10.45±0.90 17.5±0.74 68.58±1.1 48.8±2.0
IC50 > 50±1.2 > 50±1.4 2.97±0.87 5.1±0.70
*Data obtained from triplicate determinations (n=3) and shown as mean±SD. IC50 is
calculated by four parameter logistic (4PL) nonlinear regression analysis and expressed as
mean±SD (n=3).
Table 3: Cytotoxic evaluation of different fractions of A. halimus L. against MCF-7 and

PC3 carcinoma cells
Fraction No. Non Surviving Fraction* (%)±SD
(100 µg/ml) MCF-7 cells PC3 cells
1 48.07±1.20 58.21±0.99
2 47.69±01.8 48.20±0.56
3 54.61±1.32 52.3±0.19
4 56.92±1.31 44.03±0.38
5 40±0.58 49.09±0.93
6 28.46±0.90 16.12±1.1
7 37.69±0.87 11.90±0.17
mean±SD (n=3).
Table 4: Multi Dose cytotoxic evaluation of the most active fractions of A. halimus L.
against MCF-7 and PC3 carcinoma cells
Conc. Non Surviving Fraction* (%)
(µg/ml) PC3 cells (Fr-1) MCF-7 cells (Fr-4)
100 60±1.7 61.23±0.99
50.0 59.0±0.95 57.4±1.4
25.0 53.23±1.15 50.0±1.2
12.5 40.34±0.89 24.0±0.97
5.0 23.21±0.68 10.40±1.10
IC50 43.7±0.56 25.0±0.87
mean±SD (n=3).

CONCLUSION
The present botanical study on Atriplex halimus L revealed that the leaf had Kranz-type leaf
anatomy. The stem exhibits anomalous secondary growth. The root showed anomalous
secondary thickening with absence of pith. The macerated tissues examination revealed the
presence of some anatomical features such as anomocytic stomata, large cluster crystals of
calcium oxalate, and spiral xylem vessels. Quantitative measurements of different cell types
are presented as an evidence for identification. The total extract and different fractions of A.
halimus L. possess cytotoxic activity with specific selectivity against MCF-7 and PC3
carcinoma cells. To the best of our knowledge, this is the first study to address the botanical
identification of Atriplex halimus L. growing in Egypt and the possible anticancer activity of
this plant using SRB assay and the different tested cell lines.
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WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

Kabbash World Journal of Pharmacy and Pharmaceutical Sciences

Volume 5, Issue 6, 84-100 Research Article ISSN 2278 – 4357

MACROSCOPIC AND MICROSCOPIC CHARACHTERIZATION OF

Department of Pharmacognosy, Faculty of Pharmacy, Tanta University, Tanta (31527),

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KEYWORDS: Atriplex halimus L., Macroscopic, Microscopic, Cytotoxicity, MCF-7 and

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MATERIALS AND METHODS

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RESULTS AND DISCUSSION

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Figure 1: Morphological characteristics of Atriplex halimus L.

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Figure 2: Transverse section of Atriplex halimus L. leaf.

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Figure 3: Transverse section of old stem.

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Figure 4: Transverse section of middle stem

Figure 5: Transverse section of young and middle stem

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Figure 6: Transverse section of root

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Figure 7: Microscopic characterization of A. halimus L. using macerated tissues

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Table 3: Cytotoxic evaluation of different fractions of A. halimus L. against MCF-7 and

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