ANTI NOCICEPTIVE ACTIVITY OF ARGENTUM NITRICUM, STAPHYSAGRIA, IGNATIA AMARA IN MICE IN COMPARISON WITH ACETYL SALICYLIC ACID (ASPIRIN)
HTML Full TextANTI NOCICEPTIVE ACTIVITY OF ARGENTUM NITRICUM, STAPHYSAGRIA, IGNATIA AMARA IN MICE IN COMPARISON WITH ACETYL SALICYLIC ACID (ASPIRIN)
Humera Anser *1 and Rahila Najam 2
Department of Pharmacology 1,Jinnah University for Women, Karachi, Pakistan.
Department of Pharmacology 2, University of Karachi, Karachi, Pakistan.
ABSTRACT: Pain is a condition that is experienced by everybody in its life irrespective of humans or animals, reason could be due to any intense stimuli. It is a common phenomenon in vertebral animals similar to that felt by humans. This study was designed to determine theanalgesic activity of Argentum Nitricum, Staphysagria and Ignatia Amara in comparison with Acetyl salicylic acid. For this purpose the method of Eddy and Leimbach using heated plate analgesic apparatus was used. Administration of water to the control, standard drug and test homeopathic remedies to animals by oral route and the reaction time of animals were noted at 30, 60, 90, 120, 150 and 180 min interval on the hot plate. In the conclusion the study proved that the argentum nitricum possess highly significant (p≤ 0.001) analgesic activity among all test remedies and is more potent than the reference drug i.e. acetyl salicylic acid.
Keywords:
|
Argentum Nitricum, Staphysagria, Ignatia Amara, Nociception
INTRODUCTION: Everybody in his/her life experiences pain, reason could be due to any intense stimuli. It is a common phenomenon in vertebral animals similar to that felt by humans. It is an alarming signal of many disease states1. Pain is an unpleasant feeling that results due to stimulation of nociceptors. The sensory receptors transmit the sensory information to the dorsal horn of spinal cord via afferent fibers of different diameters for example Aδ fiber and C fibers. Pain senses are sent to thalamus via Aδ fiber and beyond the spinothalamic tract i.e. cerebral cortex. Pain could be physiological, neuropathic, dysfunctional or due to inflammatory response.
Ideally the treatment should be targeted at specific mechanisms rather than suppressing the pain symptoms. Alarge number of therapeutic agents are available that relieves the pain 2. Among them opioidal analgesics and NSAIDs are of importance but have various adverse effects. They are used as tools in determination of analgesia and inflammation.
The aim of this study was to discover the analgesic effects of drugs of alternative medicines with less severe adverse effects as compare to NSAIDs / opioidal drugs. Homeopathy is a system of alternative medicine which is considered as a pseudoscience 3. This system includes treatments that targets the underlie causes of diseases. This system uses minimal concentration of substances that is still capable of producing effect with no adverse effect 4.Homeopathic medicines stimulate the internal environment of body cells to cure disease itself 5.
Staphysagriais remedy that is extracted from plant source Delphinium Staphysagria, which is the specie of delphinium (larkspur) of Renunculaceae family. This plant was also known as Lice –Bane or Staversacre. Maud Grieve (1931) 6 refers this as being vermifuge, a violent emetic, and cathartic. Main constituents are alkaloidal in nature and include Delphinine (an irritant poison), Delphisine, Delphinoidine, staphysagroine, Staphysagrine. Delphinine used both internally and externally for neuralgia. It also has strychnine induced convulsion reversal ability by decreasing the spinal cord excitation. It is capable of causing spinal cord paralysis as well as death due to asphyxia Staphysagrine paralyses the motor nerves like curare 6.
PLANT OF DELPHINIUM STAPHYSAGRIA. PLANT OF STRYCHNOS IGNATII.
Another remedy Ignatia amara made from plant, Botanical name is Strychnos Ignatii. It belongs to the family N.O. Loganiaceae. Faba Ignatic and Ignatia amara (Linn) are its synonym. Its habitat is in Philippine island. The main constituents contain strychnine in high content as compare to Nux vomica, Brucine, a fixed oil, a gum resin and bassorin 6. It is used as a tonic and stimulant. This plant is a source of homeopathic remedy Ignatia amara. This remedy is used in the treatment of anxiety related problem.
Another remedy Argentum Nitricum, the source is mineral i.e. silver nitrate (AgNO3). It is indicated in conditions like vertigo with trembling, apprehensions, neuralgic pain that ends up in vomiting. The type of pain could be prosoplagia (trigeminal pain), gastralgia (abdominal pain) or chest paindue to angina pectoris. It is also used in the treatment of epilepsy. It is the best remedy in hemicranias7.
Research methodology:
For analgesic activity adult albino mice of either sexes, weighing 25-30 g were used. The animals were housed two animals in one cage in a room maintained at 25 ± 2 oC, relative humidity 55-65%and were maintained under a 12 hours light / dark cycle i.e. light on from 08.00 am to 08.00 pm in the animal house of the Department of Pharmacology, University of Karachi. The animals were given standard diet and had free access to clean and purified drinking water but the animals were deprived of food overnight, before the experiment. 10 animals were used in each group.
Animals were handled as per specification provided in Helsinki Resolution 1964 and the study was approved by our Board of Advanced Studies and Research vide Resolution No 10(50).dated: 25-10-2011, 11-11-2011 and 25-11-2011.
Analgesic activity:
Heat is used as a source of pain. Animals were placed individually on the hot plate maintain at constant temperature (52±2°C) 8 and the animal response, such as paw licking or jump response was taken as the end response. Analgesic drugs/compounds increases the reaction time. The method was first described by Eddy & Leimbach9 (A cut off period of 15 sec is observed to avoid damage to the paw) 9, 10. The hot plate test has been found to be suitable for evaluation of centrally acting analgesics 11. The thermal model of the tail flick test is considered to be spinal reflex, but could also involve higher neural structures as this method identifies mainly central analgesic 12.
Administration of the control, standard drug and test drugs to animals was through oral route and the reaction time of animals were noted at 30, 60, 90, 120, 150 and 180 min interval on the hot plate after drug administration. The method of Eddy and Leimbach using heated plate analgesic apparatus was used. Acetyl salicylic acid (300mg) was used as standard reference drug for comparison.
The dose of acetyl salicylic acid was calculated on the basis of body weight of the animals, 300 mg was an adult dose. Animals (mice) were maintained under environmentally controlled conditions in five groups (n=10) of each animals. Group one served as control(distilled water only), second group was given standard reference drug (Acetyl salicylic acid)(positive control),while third, fourth and fifth group were given Argentum nitricum, Staphysagria and Ignatia amara 0.1 ml of diluted 30 C potency respectively.
Dose calculation:
2 drops of 30C potency in 30 ml then take 1tsp out of this dilution, is an adult dose.
For 25gm mice the dose will be =30/70 x 0.025=0.01ml or 10 micro liter.
Determination of analgesic activity using Eddy’s Hot plate apparatus:
Pain reflexes were measured, the apparatus used was Hot plate analgesimeter UGO Basile instrument no 7280. The pain response time were measured after 30 minutes of drug administration. For analgesic activity the cut off time was 20 second to avoid any damage 13. The pain response can be in the form of licking of hind paw, flipping of hind paw or jumping of animal out from that enclosed arena 14.
Remedies:
We selected Argentum Nitricum, Staphysagria, Ignatia Ammara of 30 C potency. The potency selected upon literature survey which has proved that these remedies range from 30-200 C potency had shown effects in treatment of purulent ophthalmia, in acute conjunctivitis, and in ulcerated cornea, gastric ulcer 15,16 and as well as in neuralgic pain, and in hemicranias 7.
Statistical analysis:
All the data is presented as Mean ± S.D. (n=10). Statistical analysis was performed using One Way Analysis Of Variance( ANOVA) followed by post hoc test Multiple comparisons. All the analysis were performed using Social Sciences version 20 (SPSS, Inc., Chicago, IL, USA).
RESULTS:
A one-way ANOVA was conducted to determine the analgesic activity and compare these homeopathic medicines with control. Argentum Nitricum, Staphysagria, Ignatia amara were compared with Control after zero min, 30 min, 60 min, 90 min, 120 min, 150 min and after 180 minutes respectively which showed significant results as compared to controls.
Argentum Nitricum at 30 min, 60 min, 150 and 180 min showed highly significant results(maximum latency to lick the paw was 29.024±1.585 at 52º C) when compared with controls(maximum latency to lick the paw was 18.075±6.09 at 52º C) (p<0.001). Argentum Nitricum at 120 min showed moderately significant results (maximum latency to lick the paw was 22.23±5.47 at 52º C) as compared to control (maximum latency to lick the paw was 14.19±6.30 at 52º C) (p<0.01). The effect of Argentum Nitricum was insignificant at 90 min (maximum latency to lick the paw was 17.9135±8.20 at 52º C) as compared to control (maximum latency to lick the paw was 14.19±6.30 at 52º C).
Staphysagria at 60 min, 120 min showed moderately significant results (maximum latency to lick the paw was 22.198±7.716 at 52º C) when compared with controls (maximum latency to lick the paw was 14.19±6.30 at 52º C). While at 150 and 180 min showed significant results (maximum latency to lick the paw was 17.586±6.062 at 52º C) when compared with control (maximum latency to lick the paw was 11.467±4.237 at 52º C). Staphysagria showed insignificant results at 30 and 90 min (maximum latency to lick the paw was 20.771±2.753 at 52º C) when compared with control (maximum latency to lick the paw was 17.9135±8.20 at 52º C).
TABLE 1. ANALGESIC ACTIVITY OF ARGENTUM NITRICUM, STAPHYSAGRIA, IGNATIA AMARA AND ACETYL SALICYLIC ACID IN COMPARISON WITH CONTROL
Drugs | Dose
|
Pre drug
0 min |
After drugs administration Mean increase in latency(sec)
|
|||||
Control
|
0.1ml (NS) |
9.355±3.462 |
30 min
7.258±1.116 |
60min
7.1125±1.392 |
90min
17.9135±8.204 |
120min
14.1957±6.306 |
150min
18.075±6.09 |
180min
11.467±4.237 |
Argentum Nitricum | 30C
(Diluted in water) 0.1 ml |
10.059±1.69
|
10.915±1.393
(p=0.001)*** |
20.693±2.518
(p≤0.001)*** |
21.335±3.728
(IS) |
22.23±5.47
( p≤0.01)** |
29.024±1.585
(p≤0.001)*** |
21.853±5.565
(p≤0.001)*** |
Staphysagria |
30C (Diluted in water) 0.1 ml |
9.162±1.508 |
9.339±1.132 (IS) |
13.355±5.113 (p≤0.01)** |
20.771±2.753 (IS) |
22.198±7.716 (p≤0.01)** |
14.331±2.588 (p≤0.05)* |
17.586±6.062 (p≤0.05)* |
Ignatia Ammara |
30C (Diluted in water) 0.1 ml |
8.726±3.947 |
8.548±3.989 (IS)
|
15.506±4.835 (p≤0.001)*** |
13.583±6.237 (IS) |
16.488±9.006 (IS)
|
11.942±3.986 (p≤0.001)*** |
13.643±10.528 (p≤0.001)*** |
Aspirin
|
300 mg |
10.648±3.991
|
14.94±2.964 (p<0.001)*** |
20.394±8.466 (p≤0.001)*** |
16.156±3.948 (IS) |
14.576±3.254 (IS) |
14.261±2.713 (p≤0.001)***
|
13.331±1.902 (p≤0.05)* |
n=10, df=4
Values are mentioned in terms of mean ± Standard Deviation using one way ANOVA. p values are calculated from the Comparison of control versus test and standard drugs ( Argentum Nitricum, Staphysagria, Ignatia Ammara, Acetyl Salicylic acid)
*p≤0.05 significant, **p≤0.01 moderately significant, ***p≤0.001 highly significant, IS = Insignificant.
TABLE 2. ANALGESIC ACTIVITY OF ARGENTUM NITRICUM, STAPHYSAGRIA, IGNATIA AMARA IN COMPARISON WITH STANDARD (ACETYL SALICYLIC ACID
Drugs | Dose
|
Pre drug
0 min |
After drugs administration Mean increase in latency(sec)
|
|||||
Control |
0.1ml (NS) |
9.35±3.46 |
30 min
7.25±1.11 |
60min
7.11±1.39 |
90min
17.91±8.20 |
120min
14.19±6.30 |
150min
18.07±6.09 |
180min
11.46±4.23 |
Argentum Nitricum |
30C
(Diluted in water) 0.1 ml |
10.06±1.69
|
10.91±1.39 (p=0.001) ### |
20.69±2.51 (IS) |
21.33±3.72 (p<0.05) # |
22.23±5.47 ( p≤0.01) ## |
29.02±1.58 (p≤0.001) ### |
21.85±5.56 (p≤0.01) ## |
Staphysagria |
30C (Diluted in water) 0.1 ml |
9.16±1.50 |
9.34±1.132 (IS) |
13.35±5.11 (p≤0.01) ## |
20.77±2.75 (IS) |
22.19±7.71 (p≤0.01) ## |
14.33±2.58 (IS) |
17.58±6.06 (IS) |
Ignatia Ammara | 30C
(Diluted in water) 0.1 ml |
8.72±3.94 | 8.54±3.98
(p<0.001) ###
|
15.50±4.83
(p≤0.05) # |
13.58±6.23
(IS) |
16.48±9.00
(IS)
|
11.94±3.98
(IS) |
13.64±10.52
(IS) |
Aspirin
|
300 mg | 10.648±3.991
|
14.94±2.964
(p<0.001)### |
20.394±8.466
(p≤0.001)### |
16.156±3.948
(IS) |
14.576±3.254
(IS) |
14.261±2.713
(p≤0.05)#
|
13.331±1.902
(IS) |
n=10 df=4
Values are mentioned in terms of mean ± Standard Deviation using one way ANOVA .p values are calculated from the comparison of control versus test and standard drugs (Argentum nitricum, Staphysagria, Ignatia Ammara, Acetyl Salicylic acid)
#p≤0.05 significant, ##p≤0.01 moderately significant, ###p≤0.001 highly significant, IS = Insignificant
GRAPH 1.
Ignatia Amara at 60 min, 150 min and 180 min showed highly significant results (p<0.001) (maximum latency to lick the paw was 15.506±4.835 at 52º C) as compared to control(maximum latency to lick the paw was 7.1125±1.392 at 52º C). It showed insignificant results at 30, 90 and120 min (maximum latency to lick the paw was 16.488±9.006 at 52º C) as compared to control (maximum latency to lick the paw was 14.195±6.09 at 52º C).
Post Hoc Analysis by multiple comparison test showed that reaction time was increased significantly by Argentum Nitricum after 30 min and 150 min, reaction time was moderately increased after 120 min and 180 min, while after 90 min the reaction time was increased significantly as compared to Acetyl salicylic acid.
Post Hoc Analysis by multiple comparison test showed that reaction time was increased moderately significantly by Staphysagria after 60 min and reaction time was moderately increased after 120 min as compared to the standard drug Acetyl salicylic acid.
Post Hoc Analysis by multiple comparison test showed that reaction time was increased highly significantly by Ignatia Amara after 30 min and significantly increased after 60 min whereas after 90 min, 120, 150, 180 min the results were insignificant as compared to the standard drug Acetyl salicylic acid.
DISCUSSION: Pain is a debilitating and unpleasant sensory feeling that is associated with tissue damage. As per definition pain is an unwanted subjective, physical, psychological experience which results due to stimulation of sensory afferent nerve fibers either Aδ fiber and C fibers that are related to pain pathways to brain through spinal cord 17.
The hot-plate test was performed, it is preferred test because of several advantages, mainly due to the sensitivity to strong analgesics and it causes partial tissue injury and due to its reliability and validity of test has been shown even in the presence of significant impairment of motor performance. The drugs may inhibit the sensitization of nociceptors and /or inhibition of central pain receptors 18, 10. Also, this test indicates involvement of opioidal receptors 19.
The hot plate method involves thermal nociceptors stimulation also, specific nociceptors involved in thermal pain perception are thermo sensitive non selective cationic channel specifically Transient receptor potential vanilloid receptor 1 (TRPV1) and Transient receptor potential vanilloid receptor protein 1 (TRPV2) 20, 21).
TRPV1are expressed in high threshold peripheral neurons and activated at temperature excess of 42 ºC. While TRPV2 receptors are activated at temperature above 50 ºC 22. It has been reported in literature by 23 that opioidal drugs have both central and peripheral pain blocking mechanism while NSAIDs block only peripheral pain mechanism. It is well established that thermal nociceptive test are more sensitive to µ opioidal agonist or central analgesics, so according to our data it is suggestive that here µ opioidal receptors are activated and producing analgesic effect24 .
The neurotransmitter involved in pain mainly is glutamate while co transmitters involved are substance P, CGRP, BDNF etc. Other neurotransmitters Nor epinephrine, serotonin, dopamine, glycine, GABA, opioidal peptides released from inhibitory neurons are involved in pain inhibition.
The pesent study indicates that test medicines may be centrally acting. Homeopathic remedies produce significant anti nociceptive effect which may be due to blockade or release of endogenous substances that stimulate pain in nerve endings similar to paracetamol and other NSAIDs25. The similar remedy, by actually increasing the disease spontaneously provokes the patient's vital force into action to discover the causative factors of the sickness and eradicate them. The purpose of the remedy appeared to stop at this spontaneous, stimulating point. The patient's own vital force takes over then and it treats and cures the patient.
The signs and symptoms apparently are not the disease; neither are they simply the manifestation of the disease but of the continuing fight between the body's defenses and the disease forces 26
Acetyl salicylic acid has been used for the treatment of moderate pain. Acetyl salicylic acid and its metabolites decreases and inhibit the postoperative hyperalgesia mainly through inhibition of COX enzyme in the spinal cord and in the periphery also 27 with a series of complex actions28 . As per our results it is shown that all of the three test medicines have promising anti nociceptive activity and have shown efficacy in pain treatment. However both central and peripheral analgesic mechanisms had been postulated. Still a thirst left for further studies to be done in order to determine mode and site of action at molecular level for better understanding of exact mechanism.
CONCLUSION: In the conclusion the data of current study showed that Argentum Nitricum posses a marked analgesic activity among all three test medicines as well as the standard reference drug i.e. Acetyl salicylic acid i.e. the reaction time to pain in Argentum Nitricum administered mice was longer than the reaction time in mice given Acetyl salicylic acid. Next to Argentum nitricum, Staphysagria also showed analgesic activity but less in efficacy than Argentum Nitricum. Ignatia Aamara stands last in the queue according to its analgesic activity.
These medicines although not indicated or used as analgesics have potential to be used as potent analgesics and this effect requires further studies in- vitro as well as in -vivo.
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How to cite this article:
Anser Hand Najam R: Anti Nociceptive Activity of Argentum Nitricum, Staphysagria, Ignatia Amara in Mice in Comparison With Acetyl Salicylic Acid (Aspirin).Int J Pharm Sci Res2015; 6(1): 147-53.doi: 10.13040/IJPSR.0975-8232.6 (1).147-53.
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Article Information
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IJPSR
Humera Anser * and Rahila Najam
Assistant Professor, Department of Pharmacology, Faculty of Pharmacy, Jinnah University For Women. Karachi, Pakistan.
humera124@gmail.com
01 May, 2014
16 July, 2014
20 September, 2014
http://dx.doi.org/10.13040/IJPSR.0975-8232.6(1).147-53
01 January, 2015