|Year : 2015 | Volume
| Issue : 1 | Page : 44-49
Evaluation of bronchodilatory properties of fruits of Opuntia elatior Mill
Sanjay P Chauhan M. Pharm, PhD 1, Navin R Sheth2, BN Suhagia1
1 Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Dharmsinh Desai University, Nadiad, India
2 Department of Pharmaceutical Sciences, Saurashtra University, Rajkot, Gujarat, India
|Date of Submission||28-Mar-2014|
|Date of Acceptance||03-Aug-2014|
|Date of Web Publication||8-Apr-2015|
Sanjay P Chauhan
Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Dharmsinh Desai University, College road, Nadiad, 387..001, Gujarat
Source of Support: None, Conflict of Interest: None
Traditionally, fruits of Opuntia elatior Mill. were used for their immunomodulatory and antiasthmatic action. The presence of potentially active nutrients and their multifunctional properties make prickly pear a perfect candidate for the production of phytopharmaceutical products.
The objective of the present study was to evaluate the bronchodilatory properties of fruits of O. elatior Mill.
Materials and methods
The bronchodilatory properties were evaluated using bronchospasm induced by acetylcholine and histamine, anticholinergic action on isolated rat ileum, and antihistaminic action on isolated guinea pig ileum.
Result and discussion
Bronchodilating effect of fruit juice was dose dependent against spasm induced by acetylcholine and histamine. O. elatior Mill. fruits possess a significant inhibitory effect on rat and guinea pig ileum contraction through antihistaminic and antimuscarinic action.
This study suggested that fruits of O. elatior Mill. possess a significant inhibitory effect on rat and guinea pig ileum, and betacyanin, an active principle compound in prickly pear, may be responsible for the action.
Keywords: Antiasthmatic, bronchodilator, Opuntia, prickly pear
|How to cite this article:|
Chauhan SP, Sheth NR, Suhagia B N. Evaluation of bronchodilatory properties of fruits of Opuntia elatior Mill. Egypt Pharmaceut J 2015;14:44-9
|How to cite this URL:|
Chauhan SP, Sheth NR, Suhagia B N. Evaluation of bronchodilatory properties of fruits of Opuntia elatior Mill. Egypt Pharmaceut J [serial online] 2015 [cited 2020 Nov 30];14:44-9. Available from: http://www.epj.eg.net/text.asp?2015/14/1/44/154717
| Introduction|| |
Airway inflammation produces airflow limitation through acute bronchoconstriction, chronic mucus plug formation, and airway wall swelling or remodeling. These symptoms may be relieved either spontaneously or after treatment. Asthma can occur at any age . Genetic susceptibility and personal/family history of atopy along with environmental exposures produce the clinical symptomatology of asthma. These signs and symptoms are highly variable in severity and duration .
Asthma has become more common in both children and adults around the world in recent decades. The increase in the prevalence of asthma has been associated with an increase in atopic sensitization and is paralleled by similar increases in other allergic disorders such as eczema and rhinitis. The rate of asthma increases as communities adopt western lifestyles and become urbanized. With the projected increase in the proportion of the world's population, that is urban from 45 to 59% in 2025, there is likely to be a marked increase in the number of asthmatics worldwide over the next two decades. It is estimated that there may be an additional 100 million persons with asthma by 2025 . Asthma is thought to affect about 3% of the population in most countries.
The cactus Opuntia spp. (subfamily: Opuntioideae, family: Cactaceae) is a xerophytic plant producing about 200-300 species. In local parlance, cactus is called Prickly pear, Slipper thorn, Tuna (English) and has different vernacular names in India, such as Hathlo thor, Chorhthlo (Gujarati), Haththathoira, Nagphana, Nagphani (Hindi), Snuhi, Vajrakantaka, Bahushala (Sanskrit), Nagadali, Nagakkali (Tamil), Nagamulla, and Nagajemudu (Telugu). It was found that all cacti in India did not belong to one species as Opuntia dillenii, but three to four species are distributed over different regions in India. O. dillenii Haw. is found mainly in the southern parts of India, whereas Opuntia vulgaris Mill. (Syn Opuntia monacantha Haw.) is distributed mainly in the northern parts; Opuntia elatior Mill. is found in western India ,. The presence of potentially active nutrients and their multifunctional properties make Opuntia spp. fruits and cladodes perfect candidates for the production of phytopharmaceutical products. Although traditionally appreciated for its pharmacological properties by the native Americans, cactus pear is still hardly recognized because of insufficient scientific information . The Opuntia spp. were used for their analgesic and anti-inflammatory, anticancer, antidiabetic, antihyperlipidemic and antihypercholesterolemic, antioxidant, antiulcer, antiviral, diuretics, immunomodulatory, improve platelet function, neuroprotective, wound healing, monoamino-oxidase inhibitor, and other nutritional importance ,.
Little is known regarding the medicinal properties of betalains and other phytoconstituents from Opuntia spp.; however, essentially nothing is known regarding their potential immunomodulatory properties. Of the numerous Opuntia spp., bioactive compounds have been isolated and characterized primarily from Opuntia ficus-indica, Opuntia polycantha, Opuntia stricta, and O. dillenii for various medicinal properties . Although mast cells play an important role in allergy and asthma, the potential immunomodulatory effects of betalains from Opuntia spp. were not studied and reported. On the basis of previous studies and traditional use showing that prickly pear enhances immune function, we suggest that at least part of the effects of fruits of O. elatior Mill. on mast cell is through inhibition of degradation function. In the present study, we examined the effects of the fruits of O. elatior Mill. on mast cell-mediated anaphylaxis.
| Materials and methods|| |
Collection, authentication, and preparation of fruit juice (Opuntia fruit juice)
The fruits of O. elatior Mill. were collected from roadside weed near Atkot (Jasdan, Rajkot, Gujarat, India) at a latitude of 22°1′48”N, longitude of 71°12′0”E, and elevation of 193 m (633 ft) and were authenticated by Raw Materials Herbarium and Museum, National Institute of Science and Communication and Information Resources, New Delhi (NISCAIR). The herbarium (specimen voucher no. rbpmpc/museum/herbarium/07-08/01) was preserved in the museum of Department of Pharmacognosy, Smt R.B. Patel Mahila Pharmacy College, Atkot. Mature fruits of O. elatior Mill. were collected and immediately taken to the laboratory. Spines and glochides were removed from fruits by just heating on the wire gauge burner and then washed with water. The peel of the fruits was removed manually, and pulp was subjected to homogenization for 5 min using boss portable blender (Boss Appliances, Daman, India). After homogenization, fruits juice was filtered through glass filter G 4 (Borosil Glass Works Ltd, Mumbai, India), and filtered Opuntia fruit juice (OFJ) was used for mast cell degranulation test.
Identification of betalains by spectrophotometric, high-performance liquid chromatography, and liquid chromatography-mass spectroscopy analysis was performed and recently published by Chauhan et al. .
Albino Wistar rats (180-250 g body weight) and guinea pigs (350-400 g) of either sex were used for this study. They were housed at ambient temperature (22 ± 1°C), relative humidity (55 ± 5%), and 12/12-h light-dark cycle. Animals had free access to Amrut brand rat pellet diet supplied by Pranav Agro Industry (Baroda, India), and water was given ad libitum. The protocol of the experiment was approved by the Institutional Animal Ethical Committee (IAEC) as per the guidance of the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Ministry of Social Justice and Empowerment, Government of India (vide certificate no. IAEC/RBPMPC/09-10/01 dated 18/07/2009).
Bronchospasm induced by acetylcholine and histamine in guinea pigs
To screen the sensitivity of guinea pigs toward spasmogens, guinea pigs of both sex (350-400 g) were placed in a plexiglass chamber and sprayed with 0.25% histamine and 0.5% acetylcholine chloride under the average pressure of 45 ± 5 mmHg for 15 s. The time to onset of respiratory distress (preconvulsive time) during challenge with these agents was measured; the guinea pigs with preconvulsive time of more than 120 s were considered to be insensitive and discarded .
The eligible guinea pigs were randomly divided into six groups, each containing six animals. The OFJ and standard drugs were administered orally. The single dose treatments were given 1½ h before the study. The following schedule of treatment was administered.
The method of histamine challenge was same as those of screening the sensitive guinea pigs, and time for preconvulsion state was noted for each animal. After about 15 days of washout period, the same animals were given the above treatments and time for preconvulsion state was noted for 0.5% acetylcholine chloride aerosol spray .
Anticholinergic action on isolated rat ileum
Albino Wistar rats of either sex were killed by a blow to the head followed by exsanguination. A portion of ileum was removed and placed in oxygenated tyrode solution at room temperature. The connective tissue was carefully trimmed from the ileum tissue, suspended in tyrode solution at 37°C, and bubbled with 95% oxygen and 5% carbon dioxide. Intestinal segments of about 20 mm length were prepared from the terminal ileum of the rat. They were cleaned and suspended in organ bath containing tyrode solution and aerated with 95% oxygen and 5% carbon dioxide. Before the measurements, the tissue was allowed to stabilize for 30 min under a resting tension of 500 mg. The longitudinal contraction was measured according to the method of Sheth et al. . The concentration-response curve of acetylcholine was recorded using graded dose of acetylcholine until the maximum response is obtained with contact time of 30 s. Thereafter, second concentration-response curves were obtained, in the presence of the test substance in the organ bath. A volume of 1 ml of different concentrations of OFJ test solutions (10, 50, 100 μl/ml) was added directly to the organ bath 3 min before the addition of acetylcholine.
Antihistaminic action of isolated guinea pig ileum
Guinea pigs were fasted for 24 h and later were killed, and a piece of ileum was isolated. The tissues were quickly transferred to Petri dish More Detailses containing tyrode solution. The ileum was mounted in an organ bath maintained at 37°C and containing tyrode solution. A basal tension of 500 mg was applied to both the tissues throughout the experiment. After stabilization for 45 min, the tissues were exposed to graded doses of histamine, and contractions were recorded. The responses to these standard drugs were re-elicited after exposing the tissue to different concentrations of OFJ test solutions (10, 50, 100 μl/ml) for 3 min. After eliciting the response, the tissues were washed out thoroughly before proceeding for next response .
All the values are expressed as mean ± SEM. The data were analyzed by one-way analysis of variance followed by Tukey's multiple comparison tests. A level of P value less than 0.05 was considered statistically significant. A level of significance was noted and interpreted accordingly.
| Results|| |
Effect on bronchospasm induced by acetylcholine and histamine in guinea pigs
Pretreatment with OFJ (5, 10, and 15 ml/kg, orally) demonstrated significant increase (P < 0.001) and dose-dependent delayed onset of convulsion in guinea pigs due to acute bronchospasm induced by 0.25% histamine and 0.5% acetylcholine aerosol [Table 1]. The percentage increase in preconvulsion time of OFJ (15 ml/kg)-treated animal was comparable with both ketotifen (1 mg/kg) and atropine (2 mg/kg) [Table 2].
|Table 1: Effect of OFJ on preconvulsion time of guinea pig after histamine and acetylcholine aerosol exposure|
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|Table 2: Effect of OFJ on histamine and acetylcholine-induced bronchospasm in guinea pigs|
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Anticholinergic action on isolated rat ileum
Rat ileum suspended in tyrode solution with 1 g of tension stabilized for 15 min. Acetylcholine (1.83 × 10 -7 to 1.46 × 10 -6 mol/l) produced a concentration-dependent contraction of tissue, reaching its maximum within 30 s of tissue contact time. The OFJ at a dose of 50 and 100 μl/ml significantly (P < 0.001) inhibited acetylcholine-induced contraction response and caused 35 and 43% reductions to the response of 1.47 μmol/l acetylcholine, respectively. OFJ at a concentration of 10 μl/ml significantly (P < 0.05) inhibited the response of 1.1 μmol/l acetylcholine [Table 3] and [Figure 1].
|Figure 1: Effect of Opuntia fruit juice on tension development in isolated rat ileum. Values are mean ± SEM (n = 6), analyzed by one-way analysis of variance followed by Tukey's multiple comparison test (aP < 0.001; *P < 0.05, for change difference versus acetylcholi ne response).|
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|Table 3: Effect of OFJ on acetylcholine-induced percentage maximum contraction of isolated rat ileum|
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Antihistaminic action of isolated guinea pig ileum
Using the isolated guinea pig ileum as a model, the effects of OFJ (10, 50, and 100 μl/ml) on histamine-induced contractions were studied. As shown in [Table 4] and [Figure 2], histamine (1.085 × 10 -7 to 1.085 × 10 -6 mol/l) produced dose-dependent contractions of guinea pig ileum. Pretreatment with OFJ inhibited the contractions of histamine. OFJ at concentrations of 50 and 100 μl/ml had significant (P < 0.01 and P < 0.001) inhibitory effect on the histamine concentration-response curve, reducing the maximum induced contraction, and caused 16 and 55% reductions to the response of 1.085 × 10 -6 mol/l histamine.
|Figure 2: Effect of Opuntia fruit juice on tension development in isolated guinea pig ileum. Values are mean ± SEM (n = 6), analyzed by one-way analysis of variance followed by Tukey's multiple comparison test (***P < 0.001; **P < 0.01; *P < 0.05, for change versus histamine c ontractions).|
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|Table 4: Effect of OFJ on histamine-induced percentage maximum contraction of isolated guinea pig ileum|
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| Discussion|| |
As bronchodilators, mediator-release inhibitors, and anti-inflammatory drugs are the different classes of drugs used conventionally in the treatment of bronchial asthma, various animal models and experimental protocols were used in the present study to evaluate antiasthmatic activity of fruit of O. elatior Mill.
Bronchial asthma is characterized by increased airway reactivity to spasmogens . An initial event in asthma appears to be the release of inflammatory mediators (e.g. histamine, tryptase, leukotrienes, and prostaglandins). Some of these mediators directly cause acute bronchoconstriction, airway hyper-responsiveness, and bronchial airway inflammation. Spasmolytic drugs such as β-adrenergic agonists, xanthine derivatives, and anticholinergics relax the airway smooth muscles and are used as quick relief medications in acute asthmatic attacks. β-Adrenergic agonists promote bronchodilation by direct stimulation of β-adrenergic receptors in the airway smooth muscle, which lead to relaxation of bronchial smooth muscle by rapid decrease in airway resistance in vivo. Specific β2 -agonists such as salbutamol, salmeterol, etc. are used since long for symptomatic relief in asthma.
In present study, significant increase in preconvulsion time was observed due to pretreatment with fruit juice of O. elatior Mill., when the guinea pigs were exposed to either acetylcholine or histamine aerosol. This bronchodilating effect of fruit juice at high dose was comparable with ketotifen and atropine sulfate. Spasmolytic effect of O. elatior Mill. fruit was also evaluated by observing the effect of fruit juice (10, 50, and 100 μl/ml) on acetylcholine and histamine-induced ileum contractions to seek for scientific evidence for beneficial use of fruits in spasm produced by any means. The results showed antagonistic effects of the fruit juice against the contraction induced by the standard spasmogens. The results of this study indicated a rightward shift in the log dose-response curve of acetylcholine and histamine in the presence of the fruit juice of O. elatior Mill. The maximum effects of acetylcholine and histamine-induced contractions were inhibited in the presence of the fruit juice. The nonparallel rightward shift in acetylcholine and histamine log dose-response curves obtained in the presence of the fruit juice, with lowered maximum contraction effect to acetylcholine and histamine, would indicate a noncompetitive or an irreversible antagonistic effect of O. elatior Mill. fruits at muscarinic and histamine H 1 receptors . In this case, the antagonist binds irreversibly to receptor site or to another site that inhibits response to the agonist.
Control of tension in gastrointestinal smooth muscle is dependent on the intracellular Ca 2+ concentration. In general, there are two types of excitation-contraction coupling based on the type of mechanism responsible for changes in Ca 2+ concentration. Electromechanical coupling requires changes in membrane potential, which in turn activate the voltage-dependent Ca 2+ channel to trigger an influx of Ca 2+ . Acetylcholine and histamine cause contraction through specific receptors and can produce changes in tension . Both acetylcholine and histamine have functional roles in natural contraction of the gastrointestinal tract. Acetylcholine is a neurotransmitter at postganglionic parasympathetic neurons that innervate the gut. The response to acetylcholine is mediated by activation of two types (M 2 and M 3 ) of muscarinic receptors ,. Activation of these receptors results in an increase in intracellular Ca 2+ , an effect mediated by inositol triphosphate acting on internal calcium stores ,,. Serotonin (5-HT) is also an important substance in the gastrointestinal tract and is present in both enterochromaffin cells of the mucosa and neurons of the mesenteric plexus; it affects both secretion and motor activity ,. The histamine contraction is mediated by the release of acetylcholine from the cholinergic neuron as well as activation of serotonergic receptors on the smooth muscles of ileum . This experiment showed that O. elatior Mill. fruits possess a significant inhibitory effect on rat and guinea pig ileum contraction through antihistaminic and antimuscarinic action.
| Acknowledgements|| |
Conflicts of interest
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[Figure 1], [Figure 2]
[Table 1], [Table 2], [Table 3], [Table 4]