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ORIGINAL ARTICLE
Year : 2013  |  Volume : 3  |  Issue : 3  |  Page : 303-308

Anti-inflammatory and analgesic activities of Dashanga Ghana: An Ayurvedic compound formulation


1 Department of Rasa Shastra and Bhaishajya Kalpana, Institute for Post Graduate Teaching and Research in Ayurveda, Gujarat Ayurved University, Jamnagar, Gujarat, India
2 Drug Discovery Laboratory, Himalaya Health Care, Makali, Bangalore, India
3 R & D Unit, SDM Centre for Research in Ayurveda and Allied Sciences, Kuthpady, Udupi, Karnataka, India

Date of Submission28-Jun-2012
Date of Acceptance24-Jul-2012
Date of Web Publication10-Jul-2013

Correspondence Address:
Galib Ruknuddin
Department of Rasa shastra and Bhaishajya kalpana, Institute for Post Graduate Teaching and Research in Ayurveda, Gujarat Ayurved University, Jamnagar, Gujarat
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/2231-0738.114877

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   Abstract 

Background and Objectives : Inflammation is the self-protective reaction of tissues towards infection, irritants, or foreign substances. Though it is a part of host defence mechanism, when it becomes severe, it turns out to be a hopeless condition which causes damage of tissues; hence control of inflammation becomes essential. In Ayurvedic texts, a number of medicinal preparations to combat inflammation have been found; among them, Dashanga Yoga is one, which is said to be significant in combating inflammation. To revalidate this claim, the present experimental study was undertaken. Materials and Methods : Wistar strain albino rats weighing 200 ± 20 g and Swiss albino mice (26 ± 2 g) of either sex were used in the study. Pharmacologically validated models were used to evaluate anti-inflammatory and analgesic effects. Dashanga Yoga was administered at dose of 45 mg/kg and 65 mg/kg for rat and mouse, respectively. Results : Dashanga Yoga significantly inhibited carrageenan-induced paw edema (P < 0.01) at both three and six hours; however, it failed to suppress formalin-induced paw edema. It decreased the formation of granulation tissue non-significantly in the chronic inflamatory model. In analgesic activity, pretreatment with Dashanga Yoga failed to inhibit the early phase of pain, whereas moderate inhibition occurred in the late phase of pain in the formalin-induced paw-licking model. In the tail fl ick model, Dashanga Yoga significantly increased tail flick latency at 120, 180, and 240 minutes in comparison to the control group. Conclusion : The results revealed that Dashanga Yoga has anti-inflammatory and analgesic activity. Hence, it can be used in the management of pain and inflammatory conditions.

Keywords: Analgesic, anti-inflammatory, Ayurveda, Dashanga Yoga


How to cite this article:
Ruknuddin G, Biswajyoti P, Kumar PP, Krishnaiah AB, Basavaiah R. Anti-inflammatory and analgesic activities of Dashanga Ghana: An Ayurvedic compound formulation. Int J Nutr Pharmacol Neurol Dis 2013;3:303-8

How to cite this URL:
Ruknuddin G, Biswajyoti P, Kumar PP, Krishnaiah AB, Basavaiah R. Anti-inflammatory and analgesic activities of Dashanga Ghana: An Ayurvedic compound formulation. Int J Nutr Pharmacol Neurol Dis [serial online] 2013 [cited 2018 Oct 22];3:303-8. Available from: http://www.ijnpnd.com/text.asp?2013/3/3/303/114877


   Introduction Top


Inflammation has always been a problem to all clinicians and even patients. It is the self-protective reaction of the tissues towards infection, irritants, or foreign substances. Though it is a part of host defence mechanism, when it becomes severe, it turns out to be a hopeless condition which causes tissue damage. Similarly, exaggerated inflammatory reactions also harm the body. Inflammation is painful and generally causes other local manifestations. It involves a complex array of enzyme activation, mediator release, extravasations of fluid, cell migration, tissue breakdown, and repair. [1] Sometimes, it may also evoke systemic signs and symptoms such as fever, malaise, loss of appetite, and so on. Therefore control of inflammation becomes essential.

In the recent scenario, contemporary medicines have found cures for many troubling health problems including some infectious diseases that cause sickness and premature death on a large scale. These contemporary drugs offer potent treatment for several common ailments, but they also carry a high risk of adverse reactions, which in some cases become distressing. Amid all these deficits of treatment, it becomes mandatory that one should search for medicaments which can overcome all these vagaries. In this direction, a search has been made through the field of Ayurveda for medicines that can fulfill the requirements. A number of preparations to combat inflammation have been found in classical Ayurvedic texts. One such compound polyherbal formulation is Dashanga Yoga, which is said to be significant in combating inflammation. Considering this, the study is planned to revalidate the anti-inflammatory and analgesic activities of the compound in suitable experimental animals.


   Materials and Methods Top


Test formulation

The formulation composition (Dashanga Yoga) is a combination of ten herbal ingredients of equal parts [Table 1]. Drugs like Yashtimadhu, Tagara, Sukshmaila, Jatamamsi, Haridra, Daruharidra, Shirisha, and Kushta were procured from the Pharmacy, Gujarat Ayurved University, Jamnagar, India. Rakta Chandana was procured from Moodbidri, Karnataka and Hribera from Palakkad, Kerala, India. Random samples of the collected drugs were subjected to pharmacognostical studies with an intention to check their identity and genuineness. After establishing proper identity, individual ingredients were converted to coarse powder (sieve #10) after drying in the shade. All the powders were mixed together thoroughly in specified proportions to prepare a homogenous blend, which was shifted to a stainless steel container, added with specified amounts (16 parts) of water and kept aside undisturbed throughout the night. On the following day, the contents were subjected to mild heat maintaining temperature between 95 and 100°C. The contents were stirred constantly, to avoid the possibility of settling down and charring of the contents. When the volume was reduced to one-quarter (1/4), the contents were filtered through a clean cloth into a stainless steel container to obtain decoction. The decoction was further subjected to the process of reboiling with continuous stirring over a mild flame till the contents become semisolid. The semisolid contents were shifted to a tray and subjected to drying, maintaining the temperature around 50°C. The dried aqueous extract (Dashanga Ghana) was collected carefully with the help of a scraper and used in the study.
Table 1: The formulation composition of Dashanga Yoga

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Animals

Wistar strain albino rats weighing 200 ± 20 g and Swiss albino mice (26 ± 2 g) of either sex were obtained from the animal house attached to the pharmacology laboratory, Institute for Post Graduate Teaching and Research in Ayurveda (IGPT and RA), Gujarat Ayurved University, Jamnagar, Gujarat, India. The animals were maintained on 'Amrut' brand animal pellet feed of Pranav Agro Industries and tap water was given ad libitum. The temperature and humidity were kept at optimum and the animals were exposed to natural day-night cycles. The experiments were carried out in conformity with the guidelines of the Institutional Animal Ethics Committee after obtaining its permission (approval number IAEC03/08-11/PhD/02) and care was taken as per the guidelines of the Committee for the Purpose of Control and Supervision Of Experiments on Animals (CPCSEA).

Dose fixation

Dose of the test formulation was fixed by extrapolating the human dose to laboratory animals on body surface area ratio as per the table of Paget and Barnes. [2] The adult human dose (500 mg per day) was converted to animal dose. On this basis, the rat dose was found to be 45 mg/kg and 65 mg/kg for mouse. The test drug was suspended in distilled water by making a uniform suspension with 0.5% carboxymethyl cellulose (CMC) with suitable concentration depending upon the body weight of the animals and administered orally with the help of a gastric catheter sleeved to a syringe. The drug was administered to overnight-fasted animals.


   Experimental protocols Top


Anti-inflammatory activity

Carrageenan-induced paw edema in rats

The Wistar strain albino rats of either sex were weighed and randomly, divided into three groups of six each. The first group received distilled water and served as the control group. The second group received the test drug, Dashanga Ghana. The third group was administered the standard anti-inflammatory drug phenylbutazone (Wilson Laboratories, Mumbai) at a dose of 100 mg/kg. The vehicle and test drugs were administered to the respective groups for five consecutive days, whereas the standard drug was given only once, that is, one hour before the carrageenan injection. Initially, the left hind paw volumes upto the tibio-tarsal articulation were recorded prior to the carrageenan injection by using a plethysmograph. [3],[4] On the fifth day, one hour after drug administration, edema was produced by injecting 0.1 ml freshly prepared 1% carrageenan in sterile saline solution to the sub-plantar aponeurosis of the left hind limb. The rats were administered tap water at a dose of 2 ml per 100 g body weight to ensure uniform hydration and hence to minimize variations in edema formation. Paw volume was recorded three hours after carrageenan injection. Results were expressed as an increase in paw volume in comparison to the initial paw volumes and also in comparison with the control group.

Formaldehyde-induced paw edema in rats

The test conditions and groupings were similar to carrageenan-induced paw edema as mentioned above, except the standard anti-inflammatory drug used (diclofenac sodium: 5 mg/kg, Novartis India Limited). Pedal inflammation was induced by injecting 0.1 ml of 3% formaldehyde solution below the plantar aponeurosis of the right hind paw of the rats. [5] The paw volume was recorded immediately prior to compound administration (0 h) and then at 24 and 48 hours after formaldehyde injection. Results were expressed as an increase in paw volume in comparison to the initial paw volumes and also in comparison with the control group.

Cotton pellet-induced granuloma formation

The method used by Meir (1950) was adopted. [6] The selected rats were anesthetized with ether. The dorsum was shaved and swabbed with 70% (v/v) alcohol. A midline incision of 1cm was made in the intrascapular region. A small tunnel was made on either side of the incision with the help of small blunt forceps. A sterile cotton pellet weighing 100 mg was inserted per tunnel and the incision was closed with interrupted sutures after expelling the air from the tunnel. Thus, cotton pellet-inserted rats were randomly divided into three groups of six rats each. Group 1 was treated with distilled water and considered as the control group. Group 2 was administered with the test formulation, Dashanga Ghana for seven consecutive days starting from the day of implantation. The third group was taken as standard and administered the standard drug, dexamethasone (0.1 mg/kg orally, Cadila Healthcare Limited) daily for seven consecutive days. On the eighth day, the animals were anesthetized again; the cotton pellets were removed surgically and freed of extraneous tissues. The pellets were incubated at 37°C for 24 hours and dried at 60°C to constant weight. The increment in the dry weight of the pellets was regarded as a measure of granuloma formation.

Analgesic activity

Formaldehyde-induced paw licking in rats

The Wistar strain albino rats of either sex were weighed and randomly divided into three groups of six each. The first group received distilled water and served as the control group. The second group received the test drug at a dose of 45 mg/kg. Indomethacin (10 g/kg orally, Cipla Healthcare Limited) was used as the standard analgesic drug. Pain was induced by injecting 0.1 ml of 3% formalin in distilled water in the subplantar region of the right hind paw and the duration of paw licking as an index of nociception was counted in periods of 0 to 10 and 20 to 30 minutes. [7]

Tail flick test

The mice were placed on the tail flick unit so that constant heat intensity was applied to the lower third of the tail of the animal. When the animal flicked its tail in response to the noxious stimulus, both the heat source and timer were stopped. A cut-off time of 10 seconds was set to avoid damage to the tail. Thus basal reaction time of each mouse to radiant heat was recorded and those having tail flick latency (TFL) less than 10 seconds were selected. The selected mice were randomly divided into three groups of six each.The first group received a similar volume of the vehicle as the test drug and served as the normal control. The mice in group two were treated with 65 mg/kg dose of the test drug. To the third group, the standard analgesic drug, pentazocine (20 g/kg i.p., Ranbaxy Laboratories) was administered. The vehicle, test drug, and reference standards were administered to the respective groups one hour prior to the experiment. The TFL was recorded at 30, 60, 120, 180, and 240 minutes. [8]

Statistical analysis

The obtained data have been presented as mean ± SEM; the difference between the groups was statistically determined by the student's t test for unpaired data for the treated group with the level of significance set at P < 0.05.


   Results Top


The result shows [Table 2] that Ghana showed a significant reduction in the carrageenan-induced paw edema (P < 0.01) at both three and six hours in comparison to the control group. The standard anti-inflammatory drug treated group also showed a significant reduction in paw edema at both time intervals in comparison to the control group.
Table 2: Effect on carrageenan-induced paw edema

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Ghana failed to suppress the paw edema at both 24 and 48 hours after formalin injection in comparison to the control group. The standard anti-inflammatory drug-treated group showed significant inhibition in paw edema at both time intervals in comparison to the control group [Table 3].

An apparent and statistically nonsignificant decrease in granulation tissue was observed in the test formulation administered group in comparison to the control group. In the dexomethasone treated group, a significant decrease in the weight of granulation tissue was observed in comparison to the control group [Table 4].
Table 3: Effect on formaldehyde-induced paw edema

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Table 4: Effect on weight of granulation tissue

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Pretreatment with Ghana failed to inhibit the early phase of pain, whereas moderate inhibition occurred in the late phase of pain. Decrease in paw licking was also observed in the indomethacin (reference standard) treated group at both the time intervals; however, these changes were found to be significant only at the late phase of nociception [Table 5].
Table 5: Effect on formalin-induced paw-licking response

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Mice pretreated with pentazocine showed a significantly increased TFL (P < 0.05) after 30 minutes and nonsignificantly 60 minutes onwards. A significant increase in TFL was observed in the Ghana treated group at 120, 180, and 240 minutes in comparison to the control group [Table 6].
Table 6: Effect on tail flick latency

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   Discussion Top


Carrageenan induced inflammation in rats is one of the most suitable acute models to screen anti-inflammatory agents. The development of edema in the paw of the rat after injection of carrageenan is a biphasic event. The initial phase of the edema is due to the release of histamine and serotonin and the edema is maintained during the plateau phase by a kinin like substance, and the second accelerating phase of swelling is due to the release of prostaglandin like substances. [9] In the present study, pre-treatment with Ghana dosage form significantly inhibited both the phases of carargeenan-induced paw edema. Inhibition of edema observed in this model may be due to the ability of the Ghana to inhibit these chemical mediators of inflammation.

The formalin-induced inflammation in the feet of the rats may be conveniently divided into two parts, the first involving 5-hydroxytryptamine (5-HT) as the mediator and the second involving the mediator which was unrelated to 5-HT. [10] In contrast to results obtained in the carrageenan-induced paw edema, Ghana dosage form failed to suppress the paw edema at both time intervals and this shows that the test formulation did not have 5-HT suppression activity.

The formation of granulomatous tissue is related to the chronic inflammatory process which is an indication for the proliferatory phases of inflammation. Inflammation involves the proliferation of macrophages, neutrophils, and fibroblasts, which are basic sources of granuloma formation; thus, this method is widely used to evaluate the transudative and proliferative components of the chronic inflammation. The dry weight of the pellets correlates with the amount of the granulomatous tissue. [11] The Ghana decreased the weight of granulation tissue nonsignificantly, and dexamethasone which was used as the standard anti-inflammatory agent in this model decreased the weight of the granulation tissue significantly. This may indicate the ability of the test formulation in reducing the synthesis of proteins, collagen, and infiltration of macrophages.

When formalin is injected subcutaneously into the paw, it produces a reaction of intense pain. The effect is seen in two phases. The initial phase lasts for 0-10 minutes of the formaldehyde injection, and it is supposed to be mediated through the modulation of neuropeptides. [12] The second phase, which is observed by 20-30 minutes of formaldehyde injection, is supposed to be mediated through the release of inflammatory mediators like prostaglandin, and so on. Pretreatment with Ghana failed to inhibit both the early and late phases of pain.

In the tail flick model, opioid agents exert their analgesic effects via spinal receptors, which is thermal-induced nociception indicating narcotic involvement which is sensitive to the opioid μ receptor. [13] Mice pretreated with pentazocine showed an increased TFL (P < 0.05) significantly after 30 minutes and non-significantly 60 minutes onwards. Pretreatment with the Ghana increased the latency of tail flick significantly at 120, 180, and 240 minutes in comparison to the control group. The ability of Ghana to prolong the reaction latency to thermally induced pain in mice further suggests central analgesic activity. The effect observed with pentazocine was short-lived, whereas the effect observed in the Ghana was long lasting. The mechanism through which this effect is brought about may be due to the modulation of opioid receptors or by the release of endogenous analgesic factors like endorphin, and so on.

As mentioned earlier, Dashanga Ghana contains 10 different constituents and the formulation is reputed in Ayurvedic literature for its shotha hara (anti-inflammatory) effect. Further, classical literature emphasized pharmacological activities like vedana sthapana (analgesic) shotha hara (anti-inflammatory) and so on for most of the drugs. It was also scientifically proven that almost all the ingredients of this formulation have analgesic and anti-inflammatory properties like Sirisha (Albizia lebbeck) [14] and Yastimadhu (Glycyrrhiza glabra),[15] Tagara (Valeriana wallichii), [16] Rakta Chandana (Pterocarpus santalinus), [17] Haridra (Curcuma longa), [18] and Daruharidra (Berberis aristata). [19],[20] Curcumin, an alkaloid present in Haridra is found to inhibit arachidonic acid metabolism, cyclo-oxygenase, lipoxygenase, and so on, and release certain steroidal hormones. It is also found to stabilize the lysosomal membrane and cause uncoupling of oxidative phosphorylation besides having a strong free radical scavenging activity. [21] These properties may be contributed in expressing anti-inflammatory and analgesic activity of the drug.


   Conclusions Top


Dashanga Yoga Ghana has anti-inflammatory and analgesic activity. Hence it can be used in the management of pain and inflammatory conditions. However,further studies are required to identify and characterize the exact active phytoconstituents and to elucidate the exact mechanism of action, which is responsible for the observed pharmacological profile.

 
   References Top

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    Tables

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5], [Table 6]


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