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Inhibition of histamine mediated responses by Mirabilis jalapa: Confirming traditional claims made about antiallergic and antiasthmatic activity


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The roots of Mirabilis jalapa are used traditionally in allergic skin disorders and asthma. The effect of an ethanol:acetone (1:1) extract of the roots of M. jalapa was studied for antihistaminic activity using a guinea pig tracheal chain preparation and clonidine-induced mast cell granulation in mice. Its antiallergic activity was evaluated using milk-induced eosinophilia and albumin-induced paw oedema in mice. The extract (0.5 mL of 100 mg mL(-1)) inhibited histamine-induced guinea pig tracheal chain contractions non-competitively. The extract (100 or 200 mg kg(-1) i.p.) inhibited milk-induced eosinophilia, albumin-induced paw oedema and protected mast cells against clonidine-induced granulation. The study justified the folkloric use of M. jalapa in the treatment of allergic diseases and asthma.
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Inhibition of histamine mediated responses by
Mirabilis jalapa
: confirming
traditional claims made about antiallergic and antiasthmatic activity
Andrea Maxiaa; Cinzia Sannaa; Balu Salveb; Ameya Kasturec; Sanjay Kastured
a Department of Botanical Sciences, University of Cagliari, Cagliari, Italy b Sinhgad College of
Pharmacy, Pune, India c AISSMS College of Pharmacy, Pune, India d MGV's Pharmacy College, Nashik,
Online publication date: 27 October 2010
To cite this Article Maxia, Andrea , Sanna, Cinzia , Salve, Balu , Kasture, Ameya and Kasture, Sanjay(2010) 'Inhibition of
histamine mediated responses by
Mirabilis jalapa
: confirming traditional claims made about antiallergic and
antiasthmatic activity', Natural Product Research, 24: 18, 1681 — 1686
To link to this Article: DOI: 10.1080/14786410802632804
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Natural Product Research
Vol. 24, No. 18, 10 November 2010, 1681–1686
Inhibition of histamine mediated responses by Mirabilis jalapa:
confirming traditional claims made about antiallergic and
antiasthmatic activity
Andrea Maxia
, Cinzia Sanna
, Balu Salve
, Ameya Kasture
and Sanjay Kasture
Department of Botanical Sciences, University of Cagliari, Cagliari, Italy;
Sinhgad College of
Pharmacy, Pune, India;
AISSMS College of Pharmacy, Pune, India;
MGV’s Pharmacy College,
Nashik, India
(Received 1 November 2008; final version received 17 November 2008)
The roots of Mirabilis jalapa are used traditionally in allergic skin disorders and
asthma. The effect of an ethanol : acetone (1 : 1) extract of the roots of M. jalapa
was studied for antihistaminic activity using a guinea pig tracheal chain
preparation and clonidine-induced mast cell granulation in mice. Its antiallergic
activity was evaluated using milk-induced eosinophilia and albumin-induced paw
oedema in mice. The extract (0.5 mL of 100 mg mL
) inhibited histamine-
induced guinea pig tracheal chain contractions non-competitively. The extract
(100 or 200 mg kg
i.p.) inhibited milk-induced eosinophilia, albumin-induced
paw oedema and protected mast cells against clonidine-induced granulation. The
study justified the folkloric use of M. jalapa in the treatment of allergic diseases
and asthma.
Keywords: albumin; mast cells; paw oedema
1. Introduction
Allergic inflammatory responses are the cause of the symptoms of many types of diseases,
including atopic dermatitis and asthma. It is well documented that the eosinophils are of
prime importance in allergic responses (Gleich, 1990). Eosinophils are thought to
contribute to the inflammation associated with several diseases, including bronchial
asthma, allergic rhinitis and hypereosinophilic syndrome. Blood and tissue eosinophilia
are also characteristic of atopic dermatitis, drug-induced skin eruptions, eosinophilic
cellulitis and eosinophilic pustular folliculitis. Mirabilis jalapa Linn (Family:
Nyctaginaceae) is distributed as an ornamental plant throughout the tropics of the
world. Mainly it is found in Mexico, Chile, India, South America, France, Brazil and Italy.
Traditionally, in India, M. jalapa is used as a diuretic, carminative, stomachic, vermifuge,
cathartic, uterine stimulant and antispasmodic (Nadkarni, 1954). The roots of M. jalapa
also possess antiviral activity (Vivanco, Savary, & Flores, 1999). In folklore medicine it is
used in the treatment of allergic skin disorders and asthma. Therefore, we investigated
the antiallergic activity of an ethanol : acetone (1 : 1) extract of the roots of M. jalapa using
*Corresponding author. Email:
ISSN 1478–6419 print/ISSN 1029–2349 online
ß2010 Taylor & Francis
DOI: 10.1080/14786410802632804
a histamine-induced contraction of guinea pig ileum, milk-induced eosinophilia, clonidine-
induced mast cell degranulation and egg albumin-induced active paw anaphylaxis in mice.
2. Results
2.1. Evaluation of antihistaminic activity using isolated guinea pig tracheal preparation
Histamine produced a dose-dependent contraction of the isolated guinea pig tracheal
chain preparation. Mirabilis jalapa extract (MJE) significantly ( p50.05) inhibited the
histamine-induced contractions of the tracheal chain preparation. In the presence of the
extract, the agonist could not produce a maximal effect and indicated non-competitive
antagonism. MJE per se did not produce any effect on the tracheal chain. The observations
are given in Figure 1.
2.2. Milk-induced eosinophilia
In mice treated with vehicle, the difference in eosinophil counts before and 24 h after
subcutaneous injection of milk was 8.33 4.81. Subcutaneous injection of milk
(4 mL kg
) increased the number of eosinophils by 120.00 10.20. In the animals
pretreated with MJE (100 and 200 mg kg
), the difference in the eosinophil count before
and after milk was 75.0 8.35, 47.5 5.6, respectively ( p50.05), whereas the difference
in the eosinophil count in the diphenhydramine-treated group was 14.2 2.0 ( p50.001).
The observations are given in Table 1.
2.3. Clonidine-induced mast cell degranulation
The clonidine-induced mast cell degranulation was inhibited by mast cell stabiliser DSCG.
The percentage protection offered by DSCG was 85.22 6.89%. Pretreatment with MJE
0 1 2 3 4 5
Vehicle + Histamine
MJE + Histamine
(Log M)
% Contraction
(mean ± SEM)
Figure 1. Effect of MJE on histamine-induced contraction of isolated guinea pig tracheal chain
preparation. A dose response curve (DRC) of histamine in variant molar concentrations in the
absence and presence of aqueous solutions of MJE (100 mg mL
, 0.5 mL) were recorded. 1 ¼7.26,
2¼6.96, 3 ¼6.66, 4 ¼6.36; n¼5, **p50.0001, *p50.001, compared to vehicle þhistamine-treated
group by Student’s t-test (unpaired). The values are expressed as mean SEM.
1682 A. Maxia et al.
(100 and 200 mg kg
) offered significant mast cell protection to 48.75 7.68 and
66.75 10.45%.
2.4. Active paw anaphylaxis
Dexamethasone (0.5 mg kg
) and MJE (100 and 200 mg kg
) significantly ( p50.01)
decreased egg albumin-induced paw thickness from 0.82 0.02 (vehicle-treated group) to
0.28 0.03, 0.40 0.03, and 0.42 0.03 mm, respectively. Observations are given in
Table 1.
3. Discussion
The histamine released from degranulated mast cells regulates airway smooth muscle
(ASM) function in asthma (Bradding, Walls, & Holgate, 2006). In asthma patients there is
an exaggerated bronchoconstrictor response of the ASM to direct and indirect stimuli,
such as histamine and exercise. Thus targeting histamine, either by prevention of its release
from mast cells or the use of histamine receptor antagonists, becomes part of
antihistaminic therapy in asthma. The inhibition of histamine’s contractile response by
MJE indicated its ability to block the postsynaptic histamine H
receptors. The presence of
peripheral blood eosinophilia and activated eosinophils in the chronic inflammatory
infiltrate of the airways is a characteristic of both allergic and non-allergic asthma. Mast
cell-derived cytokines are known to regulate both IgE synthesis and the development
of eosinophilic inflammation (Bradding & Holgate, 1999). Eosinophilic inflammation of
the airways is correlated with the severity of asthma. These cells are likely to play a part in
the epithelial damage seen in this disease (Bousquet, Jeffery, Busse, Johnson, & Vignola,
2000). The inhibition of milk-induced eosinophilia suggests the usefulness of MJE in
treatment of eosinophilia associated with different clinical conditions.
Mast cells play a major role in the pathophysiology of asthma due to their ability to
release a host of pleiotropic autacoid mediators, proteases and cytokines in response to
activation by both immunoglobulin IgE-dependent and diverse non-immunological stimuli.
(Bradding et al., 2006; Cruse et al., 2005). Secretion of autacoid mediators such as
histamine, prostaglandin D
and leukotriene C
induces bronchoconstriction, mucus
secretion and mucosal oedema, thereby contributing to acute symptoms. Gupta et al. (1994)
Table 1. Effect of M.jalapa root extract on milk-induced eosinophilia, clonidine-induced mast cell
degranulation and egg albumin-induced paw oedema.
Treatment (mg kg
% Protection from mast
cell degranulation
Volume of paw
oedema (mL)
Vehicle 8.33 4.81 0.82 0.02
Milk 120.0 10.2
MJE (100) 75.0 8.35* 48.5 7.68** 0.40 0.03*
MJE (200) 47.5 5.6* 66.75 10.45** 0.42 0.02*
Diphen (1) 14.2 2.0**
DSCG (1 mg) 85.22 6.89**
Dexa (0.5) 0.28 0.03**
Notes: Values are mean SEM of five observations. *p50.01, **p50.001 compared to respective
control. Diphen ¼Diphenhydramine; DSCG ¼disodium cromoglycate, Dexa ¼dexamethasone.
Natural Product Research 1683
showed that clonidine releases histamine from mast cells in a similar manner to selective
liberators like compound 48/80. The MJE stabilised the mast cells in a dose-dependent way,
revealing its mast cell stabilising activity. Wu, Chun, Ebizuka and Sankawa (1991) have
reported inhibition of ovalbumin-induced paw anaphylaxis by Centipeda minima, which is
suggestive of antiallergic activity. M. jalapa also exhibited egg albumin-induced paw
oedema. Thus, the study concludes that M. jalapa has potential antihistaminic activity,
which justifies its folkloric use in the treatment of asthma and allergic skin diseases.
4. Experimental
4.1. Plant and preparation of extract
Fresh roots of M. jalapa were harvested from the Botanical Garden, Cagliari, Sardinia,
Italy in August. The plant material was authenticated at the Herbarium of the Department
of Botanical Sciences, University of Cagliari by comparison with a voucher specimen
(1423). The fresh roots (500 g) were macerated for eight days with ethanol : acetone (1 : 1).
The extract was concentrated under reduced pressure (yield: 24 g).
4.2. Animals
Albino male Swiss mice (18–25 g) and guinea pigs (275–300 g) were housed under standard
laboratory conditions (light period of 12 h per day, temperature 25 2C and humidity
55 5%) with free access to food (standard pellet chow, Lipton, India) and water
ad libitum. Food but not water was withdrawn overnight and during the experiment. The
experimental procedures were carried out in strict compliance with Institutional Animal
Ethics Committee regulations, constituted under the provisions of the Prevention of
Cruelty to Animals Act.
4.3. Drugs
Clonidine (Unichem Laboratories Pvt. Ltd, India), dexamethasone (Zydus Laboratories,
India), diazepam (Ranbaxy Laboratories, India), diphenylhydramine (Glenmark
Pharmaceuticals, India), disodium chromoglycate (DSCG) (Cipla Pharmaceuticals,
India), and histamine (Sigma, USA), were used in this study.
4.4. Reagents and chemicals
Eosin solution and white blood cell dilution fluid (Qualigens, India), toluidine blue
(Research Lab, India), RPMI buffer medium 1640 (Hi-Media, India) and other chemicals
of analytical grade were purchased from Modern Scientifics, Nashik, India.
4.5. Evaluation of antihistaminic activity using isolated guinea pig tracheal preparation
Guinea pig trachea was cut into individual rings and tied together in a series to form
a chain, as described by Goyal (1998). This was suspended in an organ bath containing
Kreb’s solution (concentration in mM L
: NaCl, 118; KCl, 4.7; CaCl
, 2.5; MgSO
, 1.2;
, 25.0; KH
, 1.2; Glucose, 11.1), maintained at 37 1C while a stream of 5%
in oxygen was bubbled through the organ tube. One end was tied to aerator tube and
the other attached to an isotonic frontal writing lever to a smoked drum. The tissue was
1684 A. Maxia et al.
allowed to equilibrate for 45 min under a load of 400 mg. A DRC for histamine
(0.5963 10
M) was taken in variant molar concentrations. A time cycle of 45 min was
followed. DRCs of histamine in the absence and presence of MJE dissolved in distilled
water were recorded. Five animals were used for the experiment.
4.6. Milk-induced eosinophilia
Mice were divided into four groups of five animals each. Blood samples were collected
from retro-orbital plexus under light ether anaesthesia. The eosinophile count was done
in each group before drug administration and 24 h after buffalo milk injection
(4 mL kg
, s.c.), as described by Brekhman and Dardymov (1969). One group received
distilled water (0.5 mL kg
i.p.) and milk. The other groups received MJE (100 and
200 mg kg
, i.p.) and diphenhydramine (1 mg kg
, i.p.). After 30 min of drug treatment
each animal was injected with milk. The difference in eosinophile count before and 24 h
after drug administration was calculated.
4.7. Clonidine-induced mast cell degranulation
Mice were divided in five groups of five animals each. A three-day drug treatment schedule
was followed. Group I received vehicle (0.1 mL, i.p.). Group II was treated with a standard
drug, i.e. DSCG (10 mgkg
i.p.). Groups III–V received MJE (100 and 200 mg kg
, i.p.
each). On the fourth day each mouse received 4 mL kg
of normal saline into the
peritoneal cavity. Peritoneal fluid was collected after 5 min of gentle massage and
transferred into siliconised test tubes containing 10 mL RPMI-1640 buffer medium (pH
7.2–7.4). This solution was then centrifuged at 400–500 rev min
for 15 min. Pellets of
mast cells were washed with the same buffer medium twice by centrifugation, discarding
the supernatant. These cells were challenged with clonidine (50 mg) and incubated at 37C
in a water bath for 10 min. This was followed by staining with 1% toluidine blue and
observed under microscope (10). A total of 100 cells were counted from different visual
areas. Percent protection against degranulation was calculated (Gupta, Srimal, Verma, &
Tandon, 1994).
4.8. Active paw anaphylaxis
Mice of either sex were sensitised by injecting 0.25 mg of egg albumin adsorbed on 6 mg of
aluminium hydroxide gel subcutaneously. On day 12 of sensitisation, the animals were
challenged with 10 mg of egg albumin in saline in the sub plantar region of the hind paw.
The contralateral paw received an equal volume of saline. The paw thickness was
measured using a micrometer 15 min after challenge. The difference in paw thickness
reflected the oedema due to antigen antibody reaction (Nair, Tamhankar, & Saraf, 1994).
Mice received vehicle, MJE (100 and 200 mg kg
, i.p.) and dexamethasone
(0.5 mg kg
, s.c.) 30 min prior to antigen challenge.
4.9. Statistical analysis
All observations are presented as mean SEM. The data was analysed by Student’s t-test
or one-way ANOVA followed by Dunnett’s test. p50.05 was considered significant.
Natural Product Research 1685
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1686 A. Maxia et al.
... This has been demonstrated in mice using clonidineinduced mast cell granulation and guinea pig tracheal chain preparation. According to the study findings, this plant's folklore use in the treatment of allergic disease and asthma is justified [108]. • An in vivo study of the immune modulatory activity of Mirabilis jalapa ethanolic tuber extracts has been included in mice. ...
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... The extract (0.5 ml of 100 mg/ml) inhibited histamine-induced Guinea pig tracheal chain contractions non-competitively. The extract (100 or 200 mg/kg, ip) also inhibited milk-induced eosinophilia, albumin-induced paw oedema and protected mast cells against clonidineinduced granulation [75]. ...
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Background: The seeds of Dolichos biflorus (DB) have been traditionally used in the treatment of cough and asthma. Aim: Based on the traditional claim, the present study was planned to evaluate the anti-allergic and anti-anaphylactic activities of DB. Materials and Methods: The ethanolic extract of the seeds of DB was prepared by cold maceration process. DB was subjected to phytochemical screening, acute toxicity studies, mast cell-stabilizing activity using compound 48/80 and anti-allergic activity using milk-induced leukocytosis and eosinophilia and passive paw anaphylaxis. Statistical Analysis: Statistical analysis was done by using one-way analysis of variance followed by Dunnett′s test. Results: The phytochemical investigation showed presence of proteins, flavonoids and glycosides. DB extract inhibited milk-induced leukocytosis and eosinophilia and also the compound 48/80 induced mast cell degranulation. DB extract significantly reduced passive paw anaphylaxis in a dose-dependent manner. Conclusion : The results demonstrated that DB extract possesses anti-allergic and anti-anaphylactic potentials that might be useful in the management of asthma.
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Two novel type I ribosome-inactivating proteins (RIPs) were found in the storage roots of Mirabilis expansa, an underutilized Andean root crop. The two RIPs, named ME1 and ME2, were purified to homogeneity by ammonium sulfate precipitation, cation-exchange perfusion chromatography, and C4 reverse-phase chromatography. The two proteins were found to be similar in size (27 and 27.5 kD) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and their isoelectric points were determined to be greater than pH 10.0. Amino acid N-terminal sequencing revealed that both ME1 and ME2 had conserved residues characteristic of RIPs. Amino acid composition and western-blot analysis further suggested a structural similarity between ME1 and ME2. ME2 showed high similarity to the Mirabilis jalapa antiviral protein, a type I RIP. Depurination of yeast 26S rRNA by ME1 and ME2 demonstrated their ribosome-inactivating activity. Because these two proteins were isolated from roots, their antimicrobial activity was tested against root-rot microorganisms, among others. ME1 and ME2 were active against several fungi, including Pythium irregulare, Fusarium oxysporum solani, Alternaria solani, Trichoderma reesei, and Trichoderma harzianum, and an additive antifungal effect of ME1 and ME2 was observed. Antibacterial activity of both ME1 and ME2 was observed against Pseudomonas syringae, Agrobacterium tumefaciens, Agrobacterium radiobacter, and others.
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The mechanism of chronic mast cell activation in asthma is unclear. Monomeric immunoglobulin (Ig)E in the absence of allergen induces mediator release from rodent mast cells, indicating a possible role for IgE in the continued activation of mast cells within the asthmatic bronchial mucosa. In this study it was investigated whether monomeric IgE induces Ca2+ influx and mediator release from human lung mast cells (HLMC). Purified HLMC were cultured for 4 weeks and then exposed to monomeric human myeloma IgE. Ratiometric Ca2+ imaging was performed on single fura-2-loaded cells. Histamine release was measured by radioenzymatic assay; leukotriene C4 (LTC4) and interleukin (IL)-8 were measured by ELISA. At concentrations experienced in vivo, monomeric IgE induced dose-dependent histamine release, LTC4 production and IL-8 synthesis. This was associated with a rise in cytosolic free Ca2+. Enhanced histamine release was still evident 1 week after initial exposure to IgE suggesting that continued exposure maintains enhanced secretion. Monomeric immunoglobulin E alone activates cultured human lung mast cells initiating Ca2+ influx, degranulation, arachidonic acid metabolism and cytokine synthesis. These findings support the hypothesis that immunoglobulin E loading of mast cells within the asthmatic airway contributes to the disordered airway physiology of this disease.
The influence of the ethanolic extract of the leaves of Vitex negundo Linn on degranulation of rat peritoneal mast cells induced by Compound 48/80 and Egg albumin was examined. The inhibitory effect of the extract was stronger in immunologically induced degranulation of mast cells. The extract was found to inhibit edema during active paw anaphylaxis in mice. It was however found to be weakly active in preventing edema induced by compound 48/80 in mice.
Coleonol, a diterpene isolated from Coleus forskohlii which directly activates the adenylate cyclase and raises intracellular cyclic AMP levels in a variety of tissues, was studied for its effect on passive cutaneous anaphylaxis (PCA) and mast cell stabilizing activity. Coleonol has shown significant anti-PCA and mast cell stabilizing activity. However it was observed that coleonol is poorly absorbed when given orally. One of its derivative, coleonol hemisuccinate has been shown to be absorbed well when given orally.
Ether, methanol and aqueous extracts of Centipeda minima (Compositae) herbs were found to have significant anti-allergy activities in passive cutaneous anaphylaxis (PCA) test. Three flavonoids, two sesquiterpene lactones and an amide were isolated from this plant material as inhibitors to induced histamine release from mast cells. The sesquiterpenes were identified as isobutyroylplenolin and senecioylplenolin by spectral investigations. The flavonoids and sesquiterpenes exhibited significant anti-allergy activity in PCA test with p.o. administration.
The mast cell is a virtual pharmacopoeia of biological substances. It used to be believed that mast cell activation was all-or-nothing, with IgE cross-linking inducing symptoms of allergy and anaphylaxis. However, the activity of mast cells in health and disease is clearly much more complicated than this. The discovery that human mast cells secrete many pleiotropic cytokines suggested there may be many novel mast cell functions, and many of these are now being realised. The ubiquitous distribution of mast cells throughout connective tissues, along epithelial surfaces, and in close proximity to blood vessels, makes their products available to a large variety of cell types including fibroblasts, glandular epithelial cells, nerves, vascular endothelial cells, smooth muscle cells, and other cells of the immune system. This tissue distribution, and the vast array of lipid mediators, proteases, proteoglycans and cytokines identified as potential products of human mast cells, explains how this interesting cell has the potential to contribute to so many diverse biological events.