Evaluation of Anti-asthmatic activity of Achyranthes aspera Linn root Extract
Shinde Ganesh S1, Rahul Kunkulol2, Sandeep Narwane2, Ravindra Jadhav3
1Department of Pharmaceutical Chemistry, Pravara Rural College of Pharmacy, Pravaranagar, India.
2Department of Pharmacology, Pravara Institute of Medical Science, Loni. India.
3Department of Pharmacognosy, Pravara Rural College of Pharmacy, Pravaranagar, India.
*Corresponding Author E-mail:
ABSTRACT:
Asthma in chronic condition is very tedious to cure and which is very common disease. The aim of study was to evaluate anti-asthmatic activity root part of. A. aspera. Collection of root part of is done from Rahata District Ahmednagar (Maharashtra). For authentication of of Achyranthes aspera A. aspera Herbarium of plant was made and sent to Biological Survey of India, Pune. The collected material was proceeded for air dry at 35-40º C and then it was pulverized in electric grinder. Extraction of obtained powder was completed in ethanol,Water-ethanol and Petroleum ether by using Soxhlet Apparatus.. The presence of Steroids, flavonides, phenolic compound, glycosides, tannins, saponins, alkaloids and carbohydrates were revealed by phytochemical screening. The extract obtained were of aerial part of screened for anti-asthmatic activity. The Petroleum ether extract of root part was result the right side shift of dose Achyranthes aspera response curve in isolated goat chain and isolated guinea pig so it is indicating antiasthmatic action of drug extract.
KEYWORDS: Asthma, Phytochemical screening, Flavonoid, Achyranthes aspera.
INTRODUCTION:
Asthma in chronic condition is very tedious to cure and which is very common disease. 17-18 million people in United States are being affected by Asthma and in the last 20 years it is found to be increased about 75%. As per the current status of asthma patients about 1 out of 13 children and 1 out of 20 adults are suffering from Asthma.1 It has been found that since 1980 number of cases of Asthma was increased for the children under the age of 5 years this alarming fact cannot be ignored. Children in school age 75% has captured by Asthma. 15-20 million asthmatic patients are estimated only in India. Data of death because of Asthma from developed countries reveals that the rate varies from 0.2-0.8 per 100,000 persons aged 6-352. Symptomatic relief is pointing requirement for curing the attack of asthma by ayurveda, unani and traditional system. One of the most elite plants is mentioned in Ayurveda and Unani system for the treatment of Asthma3.
Even in Chinese system of medicine Achyranthes aspera most essential plant is pungent, bitter, laxative, carminative, stomachic and also beneficial in the treatment of bronchitis, vomiting, piles, heart disease, ascites, itching abdominal pains, dysentery, blood diseases, dyspepsia etc5,6. So the objective of present study of A. aspera evaluate for Antiasthmatic activity
MATERIALS AND METHODS:
Plant Material:
Collection of root part of Achyranthes aspera is done from Rahata, District Ahmednagar (Maharashtra). For authentication of of Achyranthes aspera A. aspera Herbarium of plant was made and sent to Biological Survey of India, Pune. The plant botanical identification was confirmed by Dr. Priyanka Ingle with no. BSI/WRC/100-1/Tech/2019/46 Dated 22/10/2019.
Extraction:
Dried and coarsely powdered root part of A.aspera was subjected to successive solvent extraction in Soxhlet extractor using petroleum ether, Water ethanol and ethanol as solvent and the marc left was refluxed with water. All the extracts were vacuum dried to produce PEE(6.28%), AQEE(1.88%), ETE(2.94%) respectively.7
Animals:
Wistar Rat (Both male and female) weighing 150-200 gm, guinea pig (Both male and female) weighing 400-600gm were housed under standard laboratory conditions. The animals were purchased from National Institute of bioscience, Pune and were housed under 12 hrs day and night conditions for acclimatization up to one week. The animals had free access to rat food pellet (purchased from Prashant Enterprises, Pune.) The ethical committee of the institute approved the protocol of the study.
Drugs and Chemicals:
The following drugs and chemicals were used Petroleum ether AR (60-80°C) (PCL, India), ethanol AR (PCL, India), and tween 80 AR (PCL, India)
Pharmacological Studies:
1. Degranulation of rat mesenteric mast cells:
Adult male albino wistar rats were sacrificed and pieces of mesentery with connecting lobes of fat and blood vessels were rapidly dissected out and placed in Ringer Locke solution. Six petri dishes were incubated for 30 minutes. Then mesenteries were transferred to other Petri dishes containing 0.1ml of 1% w/v solution of egg albumin for 20 minutes separately. Then all these Mesenteries were transferred in 4% formaldehyde containing 0.1% toluidine blue dye and kept a side for 20 minutes. After staining and fixation of mast cells, mesenteric pieces were transferred through acetone and xylene two times and mounted on slides. Six pieces of each mesentery were used for each concentration of drug. Each piece was observed under high power light microscope (Vogel HG, 2008; Norton S, 1954). Protection of mast cell from total of at least 100 mast cells was counted.
Parameter:
Percent protection of mast cell from total of at least 100 mast cells was counted.
Percent protection of mast cells = total mast cells – degranulated mast cells.8
Table 1: Effect of root extract of Achyranthus aspera on degranulation of rat mesenteric mast cells
|
Sr. no. |
Group (n=6) |
Treatment |
% of Disrupted mast cells |
|
1 |
Control |
Aerosol of 0.5% histamine |
75.83 ± 1.16 |
|
2 |
Standard |
Received Ketotifen (20μg/ml) |
34.66 ± 0.88** |
|
3 |
Treatment -1(AAEA) |
A. aspera ethanol root extract (100μg/ml) |
54.16± 1.04* |
|
4 |
Treatment-2 (AAEAA) |
A. aspera Water-ethanol root extract (100μg/ml) |
50.00 ± 0.89* |
|
5 |
Treatment-3 (AAPEE) |
A. aspera Petroleum ether root extract (100μg/ml) |
42.16 ± 1.01** |
* P< 0.05, **P<0.01 Values are Mean ± SEM, n=6, when compared with Control by using one way ANOVA followed by Dunnette’s multiple comparison test.
2. Histamine induced bronchospasm in guinea pig:
Guinea pigs were divided into different groups (n=6) and kept in a closed chamber and exposed to an aerosol of 0.5% histamine using nebulizer. The time required to develop preconvulsive dyspnoea (PCD) was recorded.As soon as PCD commenced, animals were removed from the chamber and placed in fresh air to recover. Control group was orally treated with, 1% tween-80, at dose 10ml/kg, body weight. Test groups received oral treatment of A.Aspera root extract at doses 50, 100 and 200mg/kg body weight and salbutamol at a dose 2 mg/kg, body weight orally was administered to standard group. All the groups were given a single dose treatment for seven days. The time for the onset of PCD was recorded on day 0 before treatment and on day 7th day.9
Table 2: Effect of root extract of Achyranthes aspera on histamine induced bronchospasm in conscious guinea pigs.
|
Sr. No. |
Group (n=6) |
Treatment |
% Increase in PCT (sec) |
|
1 |
Control |
Aerosol of 0.5% histamine |
4.83 ±0.60 |
|
2 |
Standard |
Received Salbutamol (2mg/kg, p.o)+ aerosol of 0.5% histamine |
55.33±0.88** |
|
3 |
Treatment - 1(AAEA) |
A.aspera ethanol root extract (50 mg/kg) |
39.50 ±0.76* |
|
aspera ethanol root extract (100 mg/kg) |
38.40 ±0.76* |
||
|
aspera ethanol root extract (200 mg/kg) |
39.20 ±0.76* |
||
|
4 |
Treatment-2 (AAEAA) |
A.aspera water- ethanol root extract (50 mg/kg) |
43.60 ±0.76* |
|
A.aspera water- ethanol root extract (100 mg/kg) |
42.70 ±0.76* |
||
|
A.aspera water- ethanol root extract (200 mg/kg) |
43.55 ±0.76* |
||
|
5 |
Treatment-3 (AAPEE) |
A.aspera Petroleum ether root extract (50 mg/kg) |
51.86±0.94** |
|
A.aspera Petroleum ether root extract (100 mg/kg) |
51.56±0.94** |
||
|
A.aspera Petroleum ether root extract (200 mg/kg) |
52.56±0.94** |
ns- Nonsignifcant, * P< 0.05, **P<0.01 Values are Mean ± SEM, n=6, when compared with Control by using one way ANOVA followed by Dunnette’s multiple comparison test 2h after the last dose.
3. Isolated Goat tracheal chain Preperation.
Freshly isolated goat tracheal chain preparation is collected from slaughter house. The trachea was rapidly dissected free of surrounding tissues and placed in petri dishes containing oxygenated Kreb’s solution (NaCl - 114.0mM; CaCl2 - 2.5mM; KCl - 4.7mM; glucose - 11.7mM; NaHCO3 - 25mM; MgCl2 - 1.2mM; KH2PO4 - 1.2mM). Trachea was sectioned into 12 rings of about the same width and connected by means of short loops of silk thread. Tracheal chains were suspended in organ tubes filled with 20ml of kreb’s solution and equilibrated under a uniform tension of 500mg. The bathing solution was bubbled with 95% O2 and 5% CO2 at 37±100C. The tissues were equilibrated for a period of 30minutes. The PSS in organ bath was changed every 10 minutes. The responses to acetylcholine were recorded using Sherrington’s recording assembly. The effect of Chlorpheniramine and roots extracts of achyranthes aspera treatment and its interaction with contractile response was recorded.
Evaluation:
The Inhibition of contraction produced by spasmogens i.e., Acetylcholine was inhibited by Petroleum ether extract of achyranthus aspera10
4.Isolated Guinea Pig Ileum Preparation:
Guinea pigs were sacrificed using cervical dislocation method and ileum was quickly dissected out and cut into pieces. Ileum piece was suspended in organ bath containing 20ml of Tyrode’s solution and was maintained at 37±100C under basal tension of 500mg. The tissues were equilibrated for a period of 30 minutes.The responses to histamine were recorded using Sherrington’s recording assembly. The effect of Aminophyline and roots extracts of Achyranthes aspera treatment and its interaction with contractile response was recorded.
Evaluation:
The Inhibition of contraction produced by spasmogens i.e., Histamine was inhibited by Petroleum ether extract of Achyranthes aspera11
Table 3: The effect of extract of root part of A. aspera on histamine induced contractions on the isolated goat chain
|
Conc (ug/ml) |
Percentage response |
||||
|
Acetylcholine |
Standard (Chlorphenarmine) |
A. apera Ethanol extract |
A. apera Water-Ethanol extract |
A. apera Petroleum ether extract |
|
|
10 |
24.00 |
22.66 |
25.33 |
24.33 |
26.33 |
|
20 |
42.66 |
32.00 |
34.66 |
36.66 |
40.66 |
|
40 |
57.33 |
52.00 |
48.00 |
48.00 |
49.00 |
|
80 |
77.33 |
64.00 |
57.33 |
59.33 |
65.33 |
|
160 |
90.66 |
73.33 |
61.00 |
63.00 |
73.00 |
|
320 |
100 |
78.66 |
62.33 |
68.33 |
76.33 |
Table 4: The effect of extract of root part of A. aspera on histamine induced contractions on the isolated Guinea Pig Ileum Preparation
|
Conc (ug/ml) |
Percentage response |
||||
|
Histamine |
Standard (Aminophylline) |
A. apera Ethanol extract |
A. apera Water-Ethanol extract |
A. apera Petroleum ether extract |
|
|
10 |
20.31 |
18.75 |
25.33 |
24.33 |
26.33 |
|
20 |
35.93 |
32.81 |
34.66 |
36.66 |
40.66 |
|
40 |
53.12 |
48.43 |
48.00 |
48.00 |
49.00 |
|
80 |
67.18 |
65.62 |
57.33 |
59.33 |
65.33 |
|
160 |
85.93 |
75.00 |
61.00 |
63.00 |
73.00 |
|
320 |
100 |
87.50 |
62.33 |
68.33 |
76.33 |
RESULT AND DISCUSSION:
In this present study, the anti-asthmatic effects of the ethanol, water-ethanol and Petroleum ether root extract of Achyranthes aspera were evaluated. In Mast cell Degranulation model the control group showed degranulation of mast cell while groups treated with Petroleum ether extract (100μg/ml) and Ketotifen (20μg/ml) significantly (P<0.01) protected degranulation of mast cells. Histamine is one of the simple inflammatory mediator which causes bronchoconstriction and inflammation in airway pathway that also responsible for the hyperbronchial responsiveness even in immediate phase of Asthma. There was a prominent involvement of H1 receptor in comparison to H2 receptor in Asthma which was estimated by the experiment in guinea pig using respiratory smooth muscle. For brochorelaxant study at 50mg/kg, 100mg/kg and 200mg/kg of dose of root extract was given and 52.56% of protection was observed which is the greatest protection percentage in comparison with t hat of 55.33% of standard salbutamol maleate (Table 2). 200mg/kg was found to be effective (p<0.01) dose because this dose had statistical significance in post treated exposition and mean exposition time. Decease in activity was found as the amount of dose increased. Bronchial asthma is one of the chronic inflammatory diseases which cause bronchoconstriction and inflammation in airway pathway that also responsible for the hyper bronchial responsiveness to most of the stimuli likes mast cell, Tlymphocytes and eosinophils. For the contractile responses various agonists like histamine, acetylcholine, bradykinin and 5- hydroxyltrytamine are responsible The petroleum ether extract of root part of A. aspera was result the right side shift of dose response curve in isolated goat chain and isolated guinea pig so it is indicating Antiasthmatic action of drug extract.
REFERENCES:
1. P. Maheshwari, I. Somasundaram. Health Related Quality of Life Measurement in Asthma and Chronic Obstructive Pulmonary Disease. Research J. Pharm. and Tech. 2016; 9(5): 518-520. doi: 10.5958/0974-360X.2016.00097.4
2. Kim H. Mazza, J. Asthma.All Asth Clin Immun.2011;7(suppl1):1-9.https://doi.org/10.1186/1710-1492-7-S1-S2.
3. Srivastava PK. Achyranthes aspera: A potent immunostimulating plant for traditional medicine. Int J Pharm Sci Res. 2014; 5(5):1601-1611.1601-11.doi: 10.13040/IJPSR.0975-8232.5 (5).
4. Ganesh SS, Rao PS, Nandal DH and Kunkulol R.A review on pharmacological and phytochemical constituent of Achyranthes aspera Linn.IntJPharmacognosy.2021;8(2):58-64. doi link: http://dx.doi.org/10.13040/IJPSR.0975-8232.IJP.8(2).
5. Prashant D Warke, M.K. Kale.Phytopharmacognostic Study of Leaves of Achyranthes aspera Linn. Research J. Pharm. and Tech. 8(2): 2015;212-215 doi: 10.5958/0974-360X.2015.00039.6
6. Priyanka K. Shinde, Dattaprasad N. Vikhe, Ravindra S. Jadhav, Ganesh S. Shinde. Pharmacological Potential of Natural Herbs against Covid-19: A Review. Research Journal of Science and Technology. 2021; 13(4):269-4. doi: 10.52711/2349-2988.2021.00043.
7. Gokhale SB, Kokate SK and Purohit AP. A text book of pharmacognosy. Nirali Prakashan, Pune. 2006.
8. 8Sagar R.Sahoo HB. Evaluation of antiasthmatic activity of ethanolic extract of Elephantopus scaber L. leaves. Indian J Pharmacol. 2012;44(3):398-401.doi: 10.4103/0253-7613.96347.
9. Bhong Prabha N.Naikawade Nilofar.In-vitro and in-vivo Evaluation of Anti-asthmatic Activity of Eugenia jambolana bark.Research Journal of Pharmacy and Technology. 2021; 14(6):3337-2. doi: 10.52711/0974-360X.2021.00580.
10. Chilivari Alekhya, V. Alagarsamy. Anti-inflammatory and Anti-asthmatic activity of Ethanolic extract of Syzygium cumini plant bark. Research J. Pharm. and Tech. 2020; 13(11):5210-5214. doi: 10.5958/0974-360X.2020.00911.7.
11. Bhosale UA.Yegnanarayan R.Pophale P, Somani R. Effect of aqueous extracts of Achyranthes aspera Linn.on experimental animal model for inflammation.Anc Sci Life. 2012;31(4):202-6.doi: 10.4103/0257-7941.107362.
Received on 12.12.2019 Modified on 10.03.2021
Accepted on 01.09.2022 © RJPT All right reserved
Research J. Pharm. and Tech 2023; 16(5):2287-2290.
DOI: 10.52711/0974-360X.2023.00375