Therapeutic Uses and Pharmacological Properties of Plantago major L. and its Active Constituents

Abstract

In the recent years synthetic drugs have been widely replaced with herbal medicines and plant extracts because of their little undesirable and extensive beneficial effects. Plantago majorL. (also known as plantain and way bread) is a member of the Plantaginaceae family. Leaves and seeds of the plant have been widely used in folk medicine for various purposes, including treatment of an extensive range of diseases and disorders such as respiratory complications and digestive system affections. It has been also used in wound healing and as an antiinflammatory, antimicrobial and antitumor agent. Moreover, plantain contains ingredients which can neutralize internal and external poisons. Recent studies have also shown its anti-fatigue properties. Phytochemical analysis of P. major extract has indicated that this plant contains a wide range of chemicals such as polysaccharides, lipids (saturated and non-saturated), amino acids (essential and non-essential), caffeic acid derivatives, flavonoids, iridoidglycosides and terpenoids, which have the potential to exert different biological effects. Phenols (ferulic acid), flavonoids and tannins have the highest amount in Plantago leaves. The present review describes the traditional uses and recent findings(Since 2000 till date) about the pharmacological effects of Plantago major L.

Full text

J. Basic. Appl. Sci. Res., 3(9)xxxx-xxx, 2013
© 2013, TextRoad Publication
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Scientific Research
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*Corresponding author: Milad Moloudizargari, Student of Veterinary Medicine, Faculty of Veterinary Medicine, Urmia
University, Urmia, Iran. Email: miladmoludi@gmail.com Postal code: 5716114999
Phone number: +989141482523
Therapeutic Uses and Pharmacological Properties of Plantago major L. and
its Active Constituents
Ali Nazarizadeh
1
, Peyman Mikaili
2
, Milad Moloudizargari
2,
*, Shahin Aghajanshakeri
2
,
Soheil Javaherypour
2
1
Department of Pharmacology, Faculty of Pharmacy, Urmia University of Medical Sciences, Urmia, Iran
2
Student of Veterinary Medicine, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran
ABSTRACT
Inthe recent years synthetic drugs have been widely replaced with herbal medicines and plant extracts because
of their little undesirable and extensive beneficial effects. Plantago majorL. (also known as plantain and way
bread) is a member of the Plantaginaceae family. Leaves and seeds of the plant have been widely used in folk
medicine for various purposes, including treatment of an extensive range of diseases and disorders such as
respiratory complications and digestive system affections. It has been also used in wound healing and as an anti-
inflammatory, antimicrobial and antitumor agent. Moreover, plantain contains ingredients which can neutralize
internal and external poisons. Recent studies have also shown its anti-fatigue properties. Phytochemical analysis
ofP. major extract has indicated that this plant contains a wide range of chemicals such as polysaccharides,
lipids (saturated and non-saturated), amino acids (essential and non-essential), caffeic acid derivatives,
flavonoids, iridoidglycosides and terpenoids, which have the potential to exert different biological effects.
Phenols (ferulic acid), flavonoids and tannins have the highest amount in Plantagoleaves. The present review
describes the traditional uses and recent findings(Since 2000 till date) about the pharmacological effects of
Plantago major L.
KEY WORDS: Plantago major L.; Plantain; Ferulic acid; Plantaginaceae; Wound healing.
INTRODUCTION
Plantago majorL.is a member species of the Plantaginacea family. It is an herbaceous perennial with a
rosette of leaves15-30 cmin diameter. Each leaf is oval, 5-20 cm long and 4-9 cm broad, rarely up to 30 cm long
and 17 cm broad, with an acute apex and a smooth margin; there are five to nine conspicuous veins. The seeds
are quite small with an ovate shape (0.4–0.8_0.8–1.5 mm) and a slightly bitter taste (Samuelsen, 2000). The
flowers are small, greenish-brown with purple stamens, produced in a dense spike 5-15 cm long on top of a stem
13-15 cm tall (rarely to 70 cm tall) (Fig. 1). It grows better than the most of other plants in compacted soils, and
is abundant beside paths, roadsides, and other areas with frequent soil compaction. It is also common in
grasslands and as a weed in crops. It is wind-pollinated, and propagates primarily by seeds, which are held on
the long, narrow spikes which rise well above the foliage (Blamey and Grey-Wilson, 1989). The plant is native
to the most of Europe and Northern and Central Asia, and is widely naturalized elsewhere in the world, where it
is a common weed. P. majorwas spread by man from Europe throughout the world4000 years ago (Jonsson,
1983).The Indiansnamed it ‘White man’s footprint’ because it was found everywhere the Europeans had been.
This has been adapted into the genus name Plantago that is from Latin planta, meaning sole of the foot. Plantain
was also used in the time of Shakespeare and was also named in the piece "Romeo and Juliet" Act I, Scene II of
the period 1592 to 1609(Samuelsen, 2000).The plant is well known in many countries (especially in Iran) and
has been used extensively in folk medicine because of its various beneficial effects (table1).Native Americans
carried powdered roots of P. major as protection against snake bite or to ward off snake. As traditional Chinese
medicine, P. major has long been used for treating viral related disease from colds and influenza to viral
hepatitis (Chiang et al., 2002).Studies carried out on the chemical composition of the plant by various methods
(for example: simple, rapid and accurate high-performance liquid chromatography) show extensive chemical
components (tables 2). Studies conducted by Jamilahet al. (2012) on the chemical composition of various
extract (petroleum ether, methanol, ethyl acetate, n-butanol and aqueous) from P. major leaves showed that all
of them have phenol groups in their extract while having different variation of organic acid groups, flavonoids
and terpenoids. P. major leaves contain 0.07% of oleanolic acid and 0.22%of ursolic acid which are two major
terpenoids of the plant(Tarvainenet al., 2009;Samuelsen, 2000)(Fig 2).These compounds exist in almost all parts
of the plant. Bioactivity ofP. major leaves and other herbal preparations which contain these secondary
metabolites is attributed to these chemical constituents (Liu, 1995).It has been shown in one study that
ursolicacid from P. majoris a Selective Inhibitor of Cyclooxygenase-2 catalyzed prostaglandin biosynthesis;
hence, anti-inflammatory effect of the plant is possibly via this mechanism. So many other studies are found in

Mikaili et al., 2013
the literature on the pharmacological effects of these two important chemicals. Leaves of the plant are rich
sources of essential fatty acids (18:26 and 18:33)and also of carotenes (Guil-Guerrero and Rodríguez-
García, 1999).Ferulic acid which has high amounts in plantago (especially in seeds)is a hydroxyl cinnamic acid,
a type of organic compound(Mohamed et al., 2011). It is an abundant phenolic phytochemical found among
plant cell wall components (Fig.2). Ferulic acid, like many natural phenols, is an antioxidant in vitro. Recentlyin
vitro and animal model studies have suggested that ferulic acid may have direct antitumor activity against breast
and liver cancer (Valentão et al., 2001; Pierre et al., 2006).Dosage range is 3-5 g of the powdered herb 1-3 time
daily but the most common dose as infusion is 150 ml (one cupful) 3-4 time a day. All in all the plant is safe and
there are no limitations and drug contraindications reported so far; however, enough care should be taken
regarding usage of the plant in pregnant women(Zagari, 1992).
This article reviews the traditional and newly investigated uses of P. major L. along with its
pharmacological effects.
Figure 1.P. major L. (leaves and seeds).
Figure 2. Chemical structures of UA and OA
Name Chemical structure
Ursolic acid
Oleanolic acid
Ferulic acid

J. Basic. Appl. Sci. Res., 3(9)xxxx-xxx, 2013
Table 1. Traditional uses of Plantago major L. in different countries especially Iran.
Part of plant and preparation Usage Country References
Whole plant decoction
Healing different kinds of wounds such as
(snake bite, intestinal worms and infectious
wounds), cold treating,Remedy for diabetes
Colombia, Italia
Watkins et al., 2011.
Idoloet al., 2010.
Jaraldet al., 2008.
Fresh leaf of the plant
Internal inflammations such as cystitis,
enteritis and swollen abdomen
Mexico
Watkins et al., 2011.
Internal use of leaves(oral)
Respiratory catarrh;astringent effect;
bleeding, skin problems; eye inflammations;
also fresh leaves applied to treat livestock
hematomas and their skin problems;pruritus.
Colombia, Iran,
Rahimiet al., 2010.
Mir-heidari, 1994.
Zagari, 1992.
Neveset al., 2009.
Topical use of leaves (lotion)
Antipyretic,Antitussive,Emollient.Blood
rectifier, Kidney pain
Portugal, Italia, Iran
Viegiet al., 2003.
Idoloet al., 2010.
Zagari, 1992.
Mix of Leaf and Root
Anti-infective
Iran
Mir-heidari, 1994.Zagari,
1992.
Decoction and infusion of fresh leaf Kidney pain
France
Boulogne et al., 2011.
Decoction of leaves of
Plantago major
Remedy for hemorrhagic-diarrheal, Tonic,
stimulant
Central America and
Mexico
Vera-Kuaet al., 2010.
Seeds of plant
mouth inflammation
Eye inflammation
Iran, India
Mir-heidari, 1994.
Oral use of the extract
Remedy for tuberculosis
Soothing effect
Iran
Mir-heidari, 1994.
Zagari, 1992.
Decoction of root Decoction of P.
major
Anti-hemorrhagic
Remedy for pulmonary disease
Antipyretic
Iran
Mir-heidari, 1994.
Mir
-
heidari, 1994.
Decoction of leaf
with vinegar
Anti-infective
Iran
Mir-heidari, 1994.
Mixture of the sap of leaves and
honey
Remedy for Ear pain and Bruises
Iran
Mir-heidari, 1994.
Extract of the root Urinary tract infection; toothache Iran Mir-heidari, 1994.
Decoction of P. major,
Euphorbia schlechtendalii
andMelochianodiflora
Stomatitis, asthma, bronchitis, ear ache, anti-
tussive
Iran , Central America
and Mexico
Vera-Kuaet al., 2010.
Brewed leaves and root
Remedy for Ear pain
Iran
Mir
-
heidari, 1994.
Whole plant
Root of plant
Aqueous extract
Bruises,
Urinary tract, toothache
Stomatitis, Asthma, bronchitis, Ear ache
Iran
Zagari, 1992.
Zagari, 1992.
Zagari, 1992.
Juice of the plant and honey Anti tussive Iran Zagari, 1992.
Table2. Biological active compounds of P. major L. leaves and seeds (mg/g on dry weight basis)
(Mohamed
et al.
, 2011)
P. major L.
seeds
P. major L. leaves
Constituent
7.43±0.07 13.05±0.10 Total phenol (mg gallic/g)
3
.
0
3
±
.
0
3
6
.
4
1
±
0
.
0
4
Total flavonoid (mg Quercetin/g)
2.43±0.03 5.63±0.06 Tannins (mg Catechine /g)
PHARMACOLOGICAL EFFECTS
Immune Enhancing Effects
Endotoxin-free methanol extracts ofP.majorleaves, in the absence of IFN- or LPS, increased production
of nitric oxide (NO) andTNF-by rat peritoneal macrophages andstimulated lymphocyte proliferation in a dose-
dependentfashion.NO and TNF- production by untreated macrophages was negligible.The regulation of
immune parameters by the extract of P. majormay be helpful in treatment of numerous diseases (Gomez-
Floreset al., 2000).For instance, activated macrophages produce mediators of cytotoxicity such as nitric oxide
and tumour necrosis factor-alpha ( TNF-α), kinds of lymphokines which protect the host against the
development of tumors and infections by organisms such as Cryptococcus, Schistosoma, Leishmania,
Francisella, Listeria and Mycobacteria (Nathan and Hibbs, 1991;Hibbset al., 1988.)
Hepatoprotective Effects
Hepatic disorders have grown in recent years and are the cause of billions of deaths all over the world
(Williams, 2006). In one study the hepatoprotective activity of P. majorseed extract in an experimental rat
model of carbon tetrachloride (CCl
4
) induced hepatotoxicity was evaluated. Control, CCl
4
and reference groups
received isotonic saline solution, CCl
4
and silibinin, respectively. P. major groups were injected CCl
4
(0.8
ml/kg) and the extract at doses of 10, 20 and 25 mg/kg, respectively for seven days. After sacrificing animalson

Mikaili et al., 2013
the 8th day blood and liver samples were collected, and then changes in plasma marker enzymes of hepatic
damage and histopatholgical alterations were recorded. Significant decrement in alanine aminotransferase
(ALT) and aspartate aminotransferase (AST) levels was observed at the dose of (25 mg/kg) in the treated group.
Microscopic examination of the livers depicted significant recovery and minimization of necrobiotic changes, in
same group (Türelet al., 2009). Parallel study was carried out by Attaet al. (2006).The results of these studies
indicate potential hepatoprotective effect of P. major extract. Further studies may suggest replacement of
synthetic drug with this natural preparation.
Anti-ulcerogenic Effects
According to some ancient evidence, major seeds have been used as a remedy for inflammatory bowel
disease (IBD) in Iran, butthere are no or not enough studies supporting this use of the plant (Rahimiet al.,
2010).In screeninganti-ulcerogenic activity of P. major, methanol extract of the plant was assayed using ethanol
and aspirin-induced gastric ulcerationsinrat models. Leaf and seed extracts were prepared separately to observe
the difference in their pharmacological actions. In alcohol induced ulcer models, the leaf extract significantly
decreased the ulcer index with curative ratio of 87.50% but the seed extract had no considerable effect in the
same model. Ranitidine (100mg/kg) produced curative ratio of 38.90%.in aspirin-induced ulcer models. Leaf
extract at an oral dose of 400 mg/kg, significantly decreased the number of gastric ulcers; however, the seed
extract had no effect on the number of ulcers. Both extracts decreased total acidity. It was also observed that
P.major (leaves and seeds) extract at an oral dose of 400mg/kg did not affect the volume of gastric juice or its
protein content. No details have been revealed about the mechanisms of these actions(Atta et al., 2005).Also one
of the main ulcerogenic agents in digestive system is Helicobacter pylori. According to Cogoet al. (2010)
studies Plantago major is capable to inhibit H. pylori in vitro.
Antidiarrheal Effects
In a study, the effect of ethanolicextract of PlantagoLeaves was evaluated on castor oil-induced diarrhea
andgastrointestinal movements in rats (charcoal meal) and on the motility of duodenum isolated from freshly
slaughtered rabbits.P. majoratadose of 200mg/kg (oral) demonstrated significant antidiarrheal effect for at least
4h.This activity was potentiated at a dose of 400mg/kg (oral). In addition, the extract significantly decreased the
distance travelledby the charcoal meal when given at both doses (200and 400 mg/kg).The large dose of the plant
extract was slightly more effective than the small one. The extract in a concentration of1.6 mg/mL or less
produced a transient stimulation the on motility of isolated duodenum. A higher concentration produced rapid
relaxation. The initial stimulant effect may be attributed to the presence of irritant substances and may explain
the contradiction in the folkloric use of this plant. No clear details were registered about responsible ingredients
(Atta and Mouneir, 2005).Further research is, however, needed to determine what compounds are responsible
for the Antidiarrheal effect.
Antinociceptive Effects
Methanolic extracts of leaves and seeds separately were studied on acetic acid-induced writhing and tail-
flicktestin mice, to investigate their anti-nociceptive effects. Oral administration of 400 mg/kg of the seed
extract showed significant nociceptive activity against acetic acid-induced writhes with a protection of62.3%.
However, at same dosesthe protection rate of the leaf extract was only 48.8%.These values were compared to
80.5% for the standard dipyrone (50 mg/kg) which is the synthetic drug. The smaller dose (200 mg/kg) of the
plant extract did not protect animals from painful acetic acid stimulation. The leaf extract at the dose of
400mg/kgproduced significant increase in the latency to the tail response to thermal stimulation. Mild or no
effect was observed at the small dose. No detail study has been carried out so far, about the ingredients that can
induce such analgesic effect (Atta and Abo EL-Sooud, 2004).
Antioxidant and Free Radical Scavenging Effects
Free radicals contribute to more than one hundred disorders in humans including atherosclerosis, arthritis,
and ischemia and reperfusion injury of many tissues, central nervous system injury, gastritis, cancer and AIDS
(Kumpulainen and Salonen, 1999; Cook and Samman, 1996).Free radical scavengers are any compounds that
react with free radicals in a biological system, thus reducing free radical-induced damage and protecting against
the indirect effects of free radicals. Ethanolic, hot and cold water extracts of P. major leaves and seeds were
assayed for determination of free radical-scavenging activity using stable 1, 1-diphenyl-2-picryl hydrazyl radical
(DPPH)in vitro. Highest antioxidant activity was observed with the ethanolic leaf extract even at a low
concentration of 20 ppm (78% activity). In the same concentration the ethanolic seed extract had really low
activity (25%). The antioxidant activity of both mentioned extracts increased in a concentration-dependent
fashion, up to 60 ppm. The rate of antioxidant activity for both extracts was so close in a concentration of
100ppm. Hot and cold water extracts of Plantago leaves were more effective than the seed extract. It was also
observed that, the ethanolic extracts were more active than the hot and cold water extracts of the samples under

J. Basic. Appl. Sci. Res., 3(9)xxxx-xxx, 2013
investigation
(Mohamed
et al.
, 2011). In another study, methanolic extract of the whole plant was studied to
measure the antioxidant activity and determine responsible contents. Results showed that the scavenging
property against DPPH, is probably due to hydroxyl groups existing in the phenolic compounds of the plant.
The extractata concentration of 0.8 mg/g had 89.3±1.5% scavenging activity and with an IC50of 0.32mg/ml.
Totalphenol content of the plant, responsible for the scavenging activity, was measured as 31±4mg/g
(Pourmorad
et al.
, 2006).
Another suggested mechanism responsible for the scavenging activity of P. major is that
highly active flavonoids present in the plant possess a 3',4'-dihydroxy occupied B ring and/or 3-OH group which
can embrace oxygen radicals and reduce them into neutralized substances like water (Amicet al., 2003).
Anticancer Effect
In one study, methanolic extracts from seven
Plantago
speciesused in traditional medicine among
them
P. major
, were evaluated for cytotoxicactivity against three human cancer cell lines:
the
human renal
adenocarcinoma(TK-10), the human breast adenocarcinoma (MCF-7) and the human melanoma (UACC-
62) cell lines) using the sulphorhodamine B (SRB) assay
in vitro
.
P. major
and the other six
Plantago
species showed cytotoxic activity on the breast adenocarcinoma (MCF-7) and melanoma (UACC-62)
tumoral cell lines in a concentration-dependent manner at the recommended NCI (USA) doses. None of the
extracts, with the exception of
Plantagobellardii
(GI50 = 86

g/ml), showed cytotoxic activity against renal
adenocarcinoma (TK-10) cells. It is thought that the cytotoxic activity depends basically on flavonoids,
flavone and luteolin present in the extract (Gálvez
et al.
, 2003).
Another
in vitro
study was carried out on
Plantago
ethanolic, hot and cold water extracts of leaves and
seeds separately. A dose dependent inhibition was observed for all testedextracts. The ethanolic extract of
P. major
L. leaves had the greatest effect on tumor cell growth (Dead 74% ±0.35) follow by its hot water
extract of the leaves (Dead 54.6% ± 1.21)(Mohamed
et al.
, 2011)
.Luteolin-7-O- β-glucoside, the major
flavonoid found in all species of Plantago, is known to be the responsible agent for the anti-cancer activity of the
plant.The precise mechanism responsible for the cytotoxic activity of luteolin-7-O- β-glucosideis not thoroughly
understood, however it is thought that topoisomerase-mediated DNA damage is the involved mechanism.
Luteolin-7-O-β-glucoside acts as a potent DNA topoisomerase I poison as well as its aglyconluteolin. (Gálvezet
al., 2003)
Cytotoxic Activity
The cytotoxic activity of P. major methanol extract on human transformed cells: HCT-15
(colon carcinoma), SQC-UISO (cervical carcinoma), OVCAR (ovary carcinoma) and KB
(nasopharynx carcinoma) cultured in RPMI-1640 medium has been also evaluated, in vitro. The
extract (1µg/mL) was cytotoxic against the UISO and OVCAR cell lines but stimulated the
proliferation of KB cells. (Velasco- Lezamaet al., 2006).
Ina screening of anticancer effect of forty-
five Russian plants, used in folk medicine, a parallel
in vitro
study was carried out using Mouse leukemia
cells (L1210).Methanolic extract of
P.major
had 80-100% cytotoxic effect (Goun
et al.
, 2002). Similar
work was done in Vietnameseandseventy-seven medicinal plants tested for their antiproliferative activities
against human HT-1080 fibro sarcoma cells.
P. major
was not among the most active plants (Ueda, 2002).
Studies on the efficacy of hot water extract of
P. major
leaves on Ehrlich ascites tumors in male mice were
also undertaken.
The extract was most effective at a dose of
25µg/mlagainst the tumor cells. The results
show that
P. major
couldbe proposed as an effective agent in cancer prevention (Ozaslan
et al.
, 2009).
Hematopoietic Effects
Aqueous and methanolic extracts of the aerial partsof P. major were added to bone marrow and spleen cell
medium to investigate their hematopoietic potential. The results were as following:
Bone marrow cultures: The aqueous and methanolic extracts stimulated cell proliferationin similar manner
using a dose of 0.4and 0.2 gr/mL.Maximum hematopoietic activity was observed at0.1 and 0.05 g/mL doses of
the methanolicextract(Velasco-Lezama et al., 2005).
Spleen Cultures: Doses of 0.4 and 0.2 g/mL of the aqueous extract increased thecell population by 3.30- and
4.40-fold, respectively. The same concentrations of the methanolic extract increased the population by 6.25- and
4.28-fold, respectively. The increase was significantly higher in spleen cultures than in bone marrow
cultures(Velasco-Lezamaet al., 2005).This effect of P. majoron spleen as a hematopoietic organ is thought to be
the second mechanism through which the plant exerts hematopoietic effects.
Wound Healing Effects
Use of
P. major
in wound healing has a very long history. Greekphysicians described its wound
healing activity in the first century and the
leaves were used as a remedy for dog bites (Samuelsen, 2000;
Roca-Garcia, 1972).It is also well known for its wound healing property in Scandinavia
. The common
Norwegian and Swedish name for
P. major
is
groblad
meaning "healing leaves"
(Samuelsen, 2000). Hence,

Mikaili et al., 2013
the plant`s Water (fresh and dried leaves) and ethanol-based leaves extracts were studied using scratch assay
with the oral epithelial cellsin vitro to validate
it’s
ancient traditional use. Concentrations of 0.1, 1 and 10mg/ml
of the plant extracts added to cell culture media to observe cell proliferation/migration. Apart from the highest
concentration of 10mg/mL, ethanol-based extracts had the most beneficial effect, followed by water extracts of
fresh leaves, ethanol plus water extracts of dried leaves and, finally, water extracts of dried leaves. Maybe
polyphenols are the responsible compounds for wound healing. Phytochemical analysis showed that high levels
of plantamajoside and other polyphenols exist in ethanol-based extract compare with other tested extracts. Other
involved substances are polysaccharides (Zubairet al., 2012).In a whole, a mixture of antioxidants are said to be
effective the wound healing process of the plant (Yokozawaet al., 1997).
Anti-inflammatory Effects
Inflammation is a complex event linked to tissue damage whether by bacteria, physical trauma,
chemical, heat or any other phenomenon and inflammatory response is the critical protective reaction to
these kinds of injures remarked by redness, fever, oedema (swelling) and pain of involved tissue (Morais
Lima
et al.
, 2011; Levine and Reichling, 1999). Methanol extract of
P. major
L. seeds was assayed on
carrageenan-induced rat paw oedema to evaluate the anti-inflammatory activity.
P. major
showed anti-
inflammatory effect in a dose dependent fashion, but it was not more effective than indomethacin
(reference drug).Median effective dose (ED
50
)was determined to be 7.507 mg/kg (
Türelet al., 2009).
It could
be thought that inhibition of COX-2-catalyzed prostaglandin biosynthesis may be the involved mechanism
for the anti-inflammatory action (Ringbom
et al.
, 1998). Furthermore, flavonoid derivatives which are high
in
P. major
are other responsible constituents present in the plant (Middleton
et al.
, 2000;Havsteen, 2002).
Anti-fatigue
Effects
Fatigue is a condition which is marked by the feeling of
exhaustion
due to heavy physical activity and
generally can cause muscular pain. Ethanol extract of
P. major
seeds were studied on forty eight male mice
to determine its effect on physical strength. Forced swimming test and biochemical assays of blood were
carried out and marker factors were registered. According to the results, the extract increased swimming
time by increasing tissue glycogen (as energy source)and decreasing serum urea nitrogen and blood lactate
(as fatigue agents).Therefore, it is suggested that the extract possesses anti-fatigue effects and can improve
endurance exercise capacity (Mao-yeand and Li-guo, 2011).There is little evidence about this effect of
P.
major
and the precise mechanisms responsible, therefore more studies are required to be conducted in this
regard.
Pest Organism Managing Properties
An ethno medicinal survey was applied in the city of Pelotas, Brazil, with professionals and patients
in the Unified Health System (SUS), showed that the most frequent symptom reported for herbal drug
usage was infection (55.3%), and tanc¸ agem (
P. major
) was the plant mentioned to be the most often used
to treat this problem (37.3%) (Dias Oliveira
et al.
, 2012).
In order to evaluate efficacy of mayan (
a large family of American Indian)
traditional potions in
treating of infectious bowel disease
P. major
L. and thirty-eight other plants were studied
in vitro
using
bacteria including
Escherichia coli
,
Klebsiella pneumonia
,
Shigella flexneri
,
Salmonella typhi
and protozoa
such as
Entamoeba histolytica
and
Giardia lamblia
. Mixture of
Melochia nodiflora
,
Euphorbia
schlechtendali
and
P. major
was active against
Giardia lamblia
(IC
50
= 21.78

g/ml). The efficacy of this
formulation for both protozoa was comparable with positive control (metronidazole). Moreover, another
formula composed of four herbs,
Trema micrantha
,
Euphorbia schlechtendali
,
Diphysa carthagenensis
and
P. major
showed good activity against
Giardia lamblia
with an of IC
50
= 12.71

g/ml. Both formulae
of
P. major
were so close to the efficacy index of metronidazole against
Giardia lamblia
and
Entamoeba
histolytica
. These two formulae showed no considerable effect against bacteria (Vera-Ku
et al.
, 2010). In
another study, extracts of 13 Brazilian medicinal plants were screened for their antimicrobial activity
against bacteria and yeasts. Leaves extract of
P. major
presented some degrees of antibacterial activity and
was not among the most active plants (Holetz
et al.
, 2002).
Similar works were carried out by SaltanÇitoğlu and Altanlar (2003), but the results were adverse.
According to this study, when compared with the standard antibiotics,
P. major
extract was found to have
good activities against
E. coli
and
S. aureus
. However,
P. major
did not show any activity against
B.
subtilis
. Further details are available in table 3.

J. Basic. Appl. Sci. Res., 3(9)xxxx-xxx, 2013
Table3. The inhibition zones diameters of free and aqueous ethanolic extracts of P. major (mm) (Çitoglu and
Altanlar, 2003).
N.T.: not tested. (-): no inhibition zone, Ampicillin and Fluconazole are positive control.
Antibacterial effects of acetone and ethyl alcohol extracts of
P. major
L. leaves were studied, using
macro dilution liquid (tube) method. Both extracts were tested for nine bacteria species (
Bacillus cereus,
Bacillus subtilis, Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Klebsiella
pneumonia, Pseudomonas aeruginosa, Proteus mirabilis, and Salmonella enteritidis
). The ethyl alcohol
extract was only effective against
E. coli
and
B. cereus
, but acetone extract was effective on all selected
bacteria species at different concentrations (Metiner
et al.
, 2012).
Previously, parallel studies were carried out by Sharifa
et al.
(2009). The whole plant methanolic,
ethanolic and aqueous extracts of
P. major
were tested on S
taphylococcus aureus, Bacillus subtilis,
Escherichia coli, Candida albicans
and
Candida tropicalis
. The methanolic and ethanolic extracts at
concentrations of 100-200 mg/ml showed bactericidal activity against both Gram positive and Gram
negative bacteria tested. Electron microscopic observation demonstrated collapse in Gram positive bacteria
cell wall and blebs formations on Gram negative bacteria. No activity was observed against yeast by any of
the extracts.
In one study, the aqueous, methanol, chloroform and hexane extracts of the aerial parts (leaves and seeds)
of P. major were added to Escherichia coli, Bacillus subtilis and Candida albicans cultures and antibacterial
activities were observed in different ranges (Velasco-Lezama et al., 2005).
The antibacterial effect of a soluble pectin polysaccharide (PMII), isolated from the leaves of P. major
was examined against systemic Streptococcus pneumoniae serotype 6B using animal model of mice. It was
observed that PMII can have prophylactically protective effects (Hetland et al., 2000)
Antiviral activity of aqueous extract and pure compounds of P. major was assayed using herpes viruses
(HSV-1, HSV-2) and adenovirus species (ADV-3, ADV-8, ADV-11).The aqueous extract possessed only a
slight anti-herpes activity. In contrast, the pure compounds were active against selected viruses. Among them
caffeic acid exhibited the strongest activity against HSV-1 (EC
50
=15.3 g/ml, SI=671), HSV-2 (EC
50
=87.3 g/ml,
SI=118) and ADV-3 (EC
50
=14.2 g/ml, SI=727), whereas chlorogenic acid possessed the strongest anti-ADV-11
(EC
50
=13.3 g/ml, SI=301) activity. The potency relates to the presence of hydroxyl group in the chemical
structure. Compounds that contained hydroxyl groups at the R
1
and R
2
positions (caffeic acid, chlorogenic acid)
were more potent than the compounds containing one hydroxyl group at the R
1
position (ferulicacid, p-coumaric
acid). Results suggest the use of these compounds as a remedy for infections caused by these two viruses
(Chiang et al., 2002). Results are in contrast with those reported by McCutcheon et al. (1995).
According to a pilot study by Ali et al. (2004), interview with 492 informants from 13 villages indicated
that macerated, dried leaves of P. major can be used as a remedy for malaria. Screening study must be carried
out to determine antimalarial activity of P. major and also to isolate and identify the active compounds, which
may be regarded as future promising phytotherapeutics in the treatment of malaria.
Antigenotoxic Effects
Anti genotoxic property of the sap from greater plantain (P. major L.) was investigated using two
bacterial test systems (SOS chromo test and Rec assay). P. major showed sizable anti genotoxic effect in none
of the test systems. This means that P. major extract cannot be used as a dis mutagen (prevent DNA damage) or
bio anti mutagen (repair of damaged DNA) agent (Karamova et al., 2010).
External Poison Detoxification
Heavy metals such as lead are toxic for humans and animals and can cause various diseases. P. major L.
was grown hydroponically in a water medium supplemented with concentrations of lead ion under different
duration times and temperature regimes to evaluate the efficacy of lead detoxification by different parts of the
plant (roots, stems, leaves and whole plant). Roots of the plant showed the highest removal rate of lead than
other parts (Akram et al., 2007). As a concern in public health, the use of agents which can purify water and
environment from heavy metals is necessary. Therefore, P. major can act as a bio filter for the removal of
Pbcations from water.
Diameters of the inhibition zones (mm)
E.coli P.aeruginosa B. subtilis S. aureus C.albicans C.galabrata C. krusei
Plantago major
11
10
-
13
7
12
12
Ampicillin (25 fig) 12 N.T 13 15 N.T N.T N.T
Fluconazole(25 ng) N.T N.T N.T N.T 18 20 20

Mikaili et al., 2013
Toxicity Evaluation
In recent years synthetic drugs have been widely replaced with herbal medicines in both developed and
non-developed countries (Verma and Singh, 2010). Some possible reasons are: the development of new diseases
with severe complications for which there is still no appropriate treatment and the belief that herbal medicine
have less or no side effects. In addition to the belief that herbal medicine is naturally superior to synthetic drugs,
economically plants are cheaper sources of remedies (Capasso et al., 2000). Apart from advantages of herbal
drugs, their safety has not been confirmed scientifically and there are still some risks in their usage. Presence of
toxic constituents (pyrrolizidine alkaloids, saponins, cyanogenetic glycosides, etc.) is the main risk in this
regard. Evaluation of toxicity of chemical and natural products isolated from 20 plants was carried out using
Artemia salina L. (Artemiidae) as in vitro test and Swiss albino mice as in vivo test. LC
50
and LD
50
for P. major
were determined as 4.74 (µg/mL) and 182.54 (mg/kg), respectively. P. major was not among the most toxic
plants (Parra et al., 2001). Analysis of the anti-nutritional and toxic components showed low content of oxalic
acid (6736 mg) and erucic (3.45%) in P. major extract (Guilet al., 1997). Laboratory studies have reported
uterine stimulatory activity of P. major; therefore it should be only used under medical supervision during
pregnancy (Shipochliev, 1981). In conclusion, P. major l is a safe plant, with low content of toxic factors,
however some adverse reactions such as: nausea, vomiting, diarrhea, anorexia, bloating, hyper-sensitivity and
dermatitis may arise after treatment with the plant. Life threatening anaphylaxis may occur in more serious
cases, which is observed in high dose usage.
Conclusion and Further Scope
This review presents up to date findings about P. major, based on the most recent pharmacological
studies that support its traditional uses. The leaf extract is reliably nontoxic with strong hepato-protective and
wound healing activities, however data about the responsible constituents is little and further research is
required. Anti-fatigue effect of the plant is also one of the newly investigated effects of P. major that needs to be
further investigated.
Acknowledgment
We are grateful to thank Mr. H. Jafari Namin for his abundant support. Special thanks to Mr. Bagheri for
helping us in the preparation of table 1.
REFERENCES
Ali AA, AL-Homaidan AA. Removal of Lead Ions from Polluted Water Using Plantago major L. and Phaseolus
vulgaris L. Australian Journal of Basic and Applied Sciences. 2007.
Ali AAN, Al-rahwi K, Lindequist U. Some medicinal plants used in yemeni herbal medicine to treat malaria.
Afr. J. of Traditional, Complementary and Alternative Medicine. 2004; 1: 72-76.
Amic D, Davidovic-Amic D, Beslo D, Trinajstic N. Structure-Radical Scavenging Activity Relationships of
Flavonoids. CROATICA CHEMICA ACTA. 2003; 76:1 55-61.
Atta AH and Abo EL-Sooud K. The anti-nociceptive effect of some Egyptian medicinal plant extracts. Journal
of Ethnopharmacology. 2004; 95: 235-238.
Atta AH and Mouneir SM. Evaluation of Some medicinal Plant extracts for Anti diarrhoeal activity. Phytother
Res. 2005; 19: 481-485.
Atta AH, Naser SM, Mouneir SM. An ntiulcerogenic effect of some plants extracts. Natural products radiance.
2005; 4(4): 258-263.
Atta AH, Naser SM, Mouneir SM. Potential protective effect of some plant extracts against carbon tetrachloride
– induced hepatotoxicity. Afr. J. of Traditional,Complementary and Alternative Medicines. 2006; 3(3): 1-
9.
Blamey M and Grey-Wilson C. Flora of Britain and Northern Europe. 1989.
Boulogne I, Germosén-Robineau L, Ozier-Lafontaine H, Fleury M, Loranger- Merciris G. TRAMIL
ethnopharmalogical survey in Les Saintes (Guadeloupe, French West Indies): A comparative study.
Journal of Ethnopharmacology. 2011; 133(3): 1039-1050.
Chiang LC, Chiang W, Chang MY, Ng LT, Lin CC. Antiviral Activity of Plantago major Extracts and Related
Compounds in vitro. Antiviral Res. 2002; 55: 53-62.
CogoL L, Monteiro CLB, Miguel MD, Miguel OG, Cunico MM, Ribeiro ML, de Camargo ER, Kussen GMB,
Nogueira KS, Costa LMD. Anti-Helicobacter pylori activity of plant extracts traditionally used for the
treatment of gastrointestinal disorders. Journal of Microbiology. 2010; 41: 304–309.
Cook NC, Samman S. Flavonoids-chemistry, metabolism, cardio protective effects and dietary sources.
Nutritional Biochemistry. 1996; 7: 66-76.

J. Basic. Appl. Sci. Res., 3(9)xxxx-xxx, 2013
Dias Oliveira SG, Moura FRR, Demarco FF, Nascente PS, Del Pino FAB, Lund RG. An ethno medicinal survey
on phytotherapy with professionals and patients from Basic Care Units in the Brazilian Unified Health
System. Journal of Ethnopharmacology. 2012; 140: 428- 437.
Galvez M, Martin-Cordero C, Lopez-Lazaro M, Cortes F, Ayuso MJ. Cytotoxic effect of Plantago spp. on
cancer cell lines. Journal of Ethnopharmacology. 2003; 88: 125-130.
Gomez-Flores R, Calderon CL, Scheibel LW, Tamez-Guerra P, Rodriguez-Padilla C, Tamez-Guerra R, Weber
RJ. Immune enhancing Properties of Plantago major Leaf Extract. Phytother Res. 2000; 14: 617-622.
Goun EA, Petrichenko VM, Solodnikov SU, Suhinina TV, Kline MA, Cunningham G, Nguyen C, Miles H.
Anticancer and antithrombin activity of Russian plants. Journal of Ethnopharmacology 2002; 81: 337-
342.
Guil-Guerrero JL and Rodríguez-García I. Lipids classes, fatty acids and carotenes of the leaves of six
ediblewild plants. Eur Food Res Technol. 1999; 209: 313-316.
Guil JL, Rodr´ıguez-garc´ıa I, Torija E. Nutritional and toxic factors in selected wild edible plants. Plant Foods
for Human Nutrition. 1997; 51: 99–107.
Havsteen BH. The biochemistry and medical significance of the flavonoids. Pharmacology therapeutics. 2002;
96: 67-202.
Hetland G, Samuelsen AB, L∅vik M, Paulsen BS, Aaberge IS, Groeng EC, Michaelsen TE. Protective Effect of
Plantago major L. Pectin Polysaccharide against Systemic Streptococcus pneumoniae Infection in Mice.
Scandinavian Journal of Immunology. 2000; 52: 348-355.
Hibbs JB Jr, Taintor RR, Vavrin Z, Rachlin EM. Nitric oxide: a cytotoxic activated macrophage effector
molecule. Biochem Biophys Res Commun 1988; 157: 87-94.
Holetz FB, Pessini GL, Sanches NR, Cortez DAG, Nakamura CV, Filho BPD. Screening of Some Plants Used
in the Brazilian Folk Medicine forthe Treatment of Infectious Diseases. MemInstOswaldo Cruz, Rio de
Janeiro. 2002; 97(7): 1027-1031.
Idolo M,Motti R, Mazzoleni S. Ethnobotanical and phytomedicinal knowledge in a long-history protected area
the Abruzzo, Lazio and Molise National Park (Italian Apennines). Journal of Ethnopharmacology. 2010;
127: 379-395.
Jamilah J,Sharifa AA, Sharifah NRSA. GC-MS Analysis of Various Extracts from Leaf of Plantago major used
as Traditional Medicine. World Applied Sciences Journal (Towards the Traceability of Halal and
Thoyyiban Application). 2012; 17: 67-70.
Jarald E, JOSHI SB, JAIN DC. Diabetes and Herbal Medicines. Iranian journal of pharmacology and
therapeutics. 2008; 7:97-106.
Jonsson, S. Blomsterboken. Markens Urter, Lyng og Trær. Teknologisk Forlag, Oslo. 1938
Karamova NS,Fatykhova DG, Abdrakhimova Y.R, Il’inskaya ON. An Investigation of Antigenotoxic Properties
of Plant Extracts of Chelidoniummajus L., Plantago major L. and Tussilagofarfara L.
EkologicheskayaGenetika. 2010; 8(2): 56-65.
Kumpulainen JT, Salonen JT. Natural antioxidants and anticarcinogens in nutrition, health and disease. The
Royal Society of Chemistry. 1999; 178- 187.
Levine JD, Reichling DB. Peripheral mechanisms of inflammatory pain. Textbook of pain, fourth edition
London:Churchilllivinstone. 1999; 59-84.
Liu J. Pharmacology of oleanic acid and ursolic acid. Journal of Ethnopharmacology. 1995; 49: 57-68.
Mao-ye W and Li-guo A. Effects of Plantago major L. seeds extract on endurance exercise capacity in mice.
Journal of Medicinal Plants Research. 2011; 5(9): 1659-1663.
McCutcheon AR, Roberts TE, Gibbons E, Ellis SM,Babiuk LA, Hancock REW, Tower GHN. Antiviral
screening of British Colombian medicinal plants. Journal of Ethnopharmacoly. 1995; 49: 101–110.
Metiner K, ÖZKANO, Seyyal AK. Antibacterial Effects of Ethanol and Acetone Extract of Plantago major L.
on Gram Positive and Gram Negative Bacteria. Kafkas Univ Vet Fak Derg. 2012; 18(3): 503-505.
Middleton EJR, Kandaswami C, Theoharides TC. The effects of plant flavonoids on mammalian cells:
implication for inflammation, heart disease, and cancer. Pharmacology review. 2000; 52: 673-751.
Mir-heidariH. Herbal encyclopedia. Office of Islamic publication Iran. 1994; 1(6): 336-339.
Mohamed, Kobeasy I, Osama, Abdel-Fatah M, Samiha M, El-Salam A, Mohamed Z. Biochemical studies on
Plantago major L. and Cyamopsistetragonoloba L. International Journal of Biodiversity and
Conservation. 2011; 3(3): 83-91.
Morais Lima GR, Albuquerque Montenegro C, Almeida CLF, Athayde-Filho PF, Barbosa-Filho JM, Batista
LM. Database Survey of Anti-Inflammatory Plants in South America: A Review. International Journal of
Molecular Sciences. 2011; 12: 2692-2749.
Nathan CF, Hibbs JB Jr. Role of nitric oxide synthesis in macrophage antimicrobial activity. Curr Op Immunol
1991; 3: 655-670.

Mikaili et al., 2013
Neves JM, Matosa C, Moutinhoa C, Queirozd G, Gomes LR. 2009. Ethnopharmacological notes about ancient
uses of medicinal plants in Trás-os-Montes (northern of Portugal). Journal of Ethnopharmacology. 2009;
124: 270–283.
Ozaslan M, Karagoz ID, Kiliç IH, Cengiz B, Kalender ME, Güldür ME, Karagoz A, Zümrütdal ME. Effect of
Plantago major sap on Ehrlich ascites tumoursin mice. African Journal of Biotechnology. 2009; 8(6):
955-959.
Parra AL, Yhebra RS, Sardiñas IG, Buela LI. Comparative study of the assay of Artemiasalina L. and the
estimate of the medium lethal dose (LD50 value) in mice, to determine oral acute toxicity of plant
extracts. Phytomedicine, Urban& Fischer Verlag. 2001; 8(5): 395–400.
Pierre G, McKinnon, Carole M. Effect of wheat variety, farming site, and bread-baking on total phenolics.
International Journal of Food Science and Technology. 2006; 41 (3): 329.
Pourmorad F, Hosseinimehr SJ, Shahabimajd N. Antioxidant activity, phenol and flavonoid contents of some
selected Iranian medicinal plants. African Journal of Biotechnology 2006; 5(11): 1142-1145.
Rahimi R, Shams-Ardekani MR, Abdollahi M. A review of the efficacy of traditional Iranian medicine for
inflammatory bowel disease. World J Gastroenterol 2010; 16(36): 4504-4514.
Ringbom T, Segura L, Noreen Y, Perera P, Bohlin L. Ursolic Acid from Plantago major, a Selective Inhibitor
of Cyclooxygenase-2 Catalyzed Prostaglandin Biosynthesis. Journal of Natural Products 1998; 61(10):
1212-1215.
Roca-Garcia H. Weeds: a link with the past. Arnoldia 1972; 30: 23–24.
SaltanÇitoğlu G and Altanlar N. Antimicrobial activity of some plants used in folk medicine. Ankara Ecz. Fak.
Derg 2003; 32(3): 159-163.
Samuelsen AB. The traditional uses, chemical constituents and biological activities of Plantago major L. A
review. Journal of Ethnopharmacology 2000; 71: 1-21.
Sharifa AA, Neoh YL, Iswadi MI, Khairul O, Abdul Halim M, Jamaludin M, MohamedAzman AB, Hing HL
(2008). Effects of Methanol, Ethanol and Aqueous Extract of Plantago major on GramPositive Bacteria,
Gram Negative Bacteria and Yeast. Annals of microscopy. 8: 42-44.
Shipochliev T. Uterotonic action of extracts from a group of medicinal plants. Vet Med Nauki. 1981; 18(4): 94-
98.
Tarvainen M, Suomela JP, Kallio H, Yang B. Triterpene Acids in Plantago major: Identification, Quantification
and Comparison of Different Extraction Methods. Chromatographia 2010; 3/4: 279-284.
Türel I, Özbek H, Erten R, Öner AC, Cengiz N, Yilmaz O. Hepatoprotective and anti-inflammatory activities of
Plantago major L. Indian J Pharmacol 2009; 41(3): 120–124.
Ueda J,Tezuka Y, Banskota AH, Tran QL, Tran QK, Harimaya Y, Saiki I, Kadota S. Antiproliferative Activity
of Vietnamese Medicinal Plants. BiolPharm Bull 2002; 25(6): 753-760.
Valentão P, Fernandes E, Carvalho F, Andrade PB, Seabra RM, Bastos ML. Antioxidant Activity of
CentauriumerythraeaIn fusion Evidenced by Its Superoxide Radical Scavenging and Xanthine Oxidase
Inhibitory Activity". Journal of Agricultural and Food Chemistry 2001; 49(7): 3476–9.
Vera-Ku M, Méndez-González M, Moo-Puc R, Rosado-Vallado M, Simá-Polanco P, Cedillo-Rivera R, Peraza-
Sánchez SR. Medicinal potions used against infectious bowel diseases in Mayantraditional medicine.
Journal of Ethnopharmacology 2010; 132: 303-308.
Velasco-Lezama R, Tapia-Aguilar R, Rom´an-Ramos R, Vega-Avila E, P´erez-Guti´errez MS. Effect of
Plantago major on cell proliferation in vitro. Journal of Ethnopharmacology 2006; 103: 36-42.
Verma S and Singh SP. Current and future status of herbal medicines. Veterinary World 2010; 1: 347-350.
Viegi L, Pieroni A, Guarrera PM, Vangelisti R. A review of plants used in folk veterinary medicine in Italy as
basis for a databank. Journal of Ethnopharmacology 2003; 89: 221-244.
Watkins F,Pendry B,Corcoran O, Sanchez-Medina A. Drug Discovery Today. 2011. Volume16, Numbers
23/24.
Williams R. Global challenges in liver disease. Hepatology 2006; 44: 521-526.
Yokozawa T, Dong E, LiuZW, Shimizu M. Antioxidant activity of flavonols in vitro. Phytotherapy Research
1997; 11: 446-449.
Zagari A 1992. Medicinal Plants. Iran Book, Teheran University Publications. 1819/4:969.
Zubair M, Ekholm A, Nybom H, RenvertS,Widen C, Rumpunen K. 2012. Effects of Plantago major L. leaf
extracts on oral epithelial cells in a scratch assay. Journal of Ethnopharmacology 141: 825– 830.