|Year : 2012 | Volume
| Issue : 2 | Page : 100-104
Anti-edematogenic and analgesic activities of Ficus benghalensis
Manoj S Mahajan, Vishal S Gulecha, Rakesh A Khandare, Aman B Upaganlawar, Hemant H Gangurde, Chandrashekhar D Upasani
Department of Pharmacology, S. S. D. J. College of Pharmacy, Neminagar, Chandwad, Maharashtra, India
|Date of Submission||21-Mar-2011|
|Date of Acceptance||14-Jun-2011|
|Date of Web Publication||9-May-2012|
Manoj S Mahajan
Department of Pharmacology, SNJB'S SSDJ College of Pharmacy, Neminagar, Chandwad, Dist. Nashik 423 101
Source of Support: None, Conflict of Interest: None
| Abstract|| |
This work was done for the investigations of anti-inflammatory and analgesic activities of methanolic extract of dried leaves of Ficus benghalensis Linn. by oral administration at doses of 10, 20 and 100 mg/kg of body weight to healthy animals. The extract was studied for anti-inflammatory activity by using carrageenan-induced hind paw edema in rats. The mean increases in paw volume and % inhibition in the edema were measured plethysmometrically at different time intervals after carrageenan (1% w/v) injection. The extract was also evaluated for analgesic activity using acetic acid-induced writhing and Eddy's hot plate method in albino mice. The methanolic extract of F. benghalensis showed significant (P<0.05) reduction in carrageenan-induced paw edema and analgesic activity evidenced from decreased acetic acid-induced writhings and increased reaction time in Eddy's hot plate method. Results of present studies suggest that methanolic extract of F. benghalensis possesses significant (P<0.05) anti-inflammatory and analgesic activities.
Keywords: Anti-inflammatory, analgesic, carrageenan-induced paw edema, Ficus benghalensis
|How to cite this article:|
Mahajan MS, Gulecha VS, Khandare RA, Upaganlawar AB, Gangurde HH, Upasani CD. Anti-edematogenic and analgesic activities of Ficus benghalensis. Int J Nutr Pharmacol Neurol Dis 2012;2:100-4
|How to cite this URL:|
Mahajan MS, Gulecha VS, Khandare RA, Upaganlawar AB, Gangurde HH, Upasani CD. Anti-edematogenic and analgesic activities of Ficus benghalensis. Int J Nutr Pharmacol Neurol Dis [serial online] 2012 [cited 2020 Aug 13];2:100-4. Available from: http://www.ijnpnd.com/text.asp?2012/2/2/100/95936
| Introduction|| |
Inflammation is considered as a primary physiologic defence mechanism that helps body to protect itself against infection, burn, toxic chemicals, allergens, or other noxious stimuli. An uncontrolled and persistent inflammation may act as an etiologic factor for many chronic illnesses. Although it is a defence mechanism, the complex events and mediators involved the inflammatory reaction can induce, maintain, or aggravate many diseases.  Inflammation has become the focus of global scientific research because of its implication in virtually all human and animal ailments. As a result of adverse effects such as gastric lesions caused by nonsteroidal anti-inflammatory drugs (NSAIDs), tolerance, and dependence induced by opiates, the use of these drugs as anti-inflammatory and analgesic agents has not been successful in all cases.  Therefore, new anti-inflammatory and analgesic drugs lacking these side effects are being researched as alternatives to NSAIDs and opiates. Attention is being focused on the investigation of the efficacy of plant-based drugs used in the traditional medicine because they are cheap and have little side effects. According to World Health Organization, about 80% of the world population use some form of herbal remedies. In India, the traditional Ayurvedic medicine uses over 1200 different herbs; herbal medicine is regularly used by about 65% of the population. Moreover, increasing interest has forced the researchers to scientifically screen various traditional medicines as these traditional claims require scientific support. This study is therefore carried out to fathom the potential of one of the potential plant Ficus benghalensis. ,
F. benghalensis 0(Moraceae, Mulberry family) is commonly known as Banyan tree or Vata or Vada in Ayurveda. This tree is considered to be sacred in many places in India. Earlier, glucoside, 20-tetratriaconthene-2-one, 6-heptatriaconthene-10-one, pentatriaconthene-5-one, β sitosterol, α-D-glucose, and meso-inositol have been isolated from the bark of the F. benghalensis.  The fruit extract exhibited antitumor activity in the potato disc bioassay. The leaves contain 9.63% crude protein, 26.84% crude fibres, 2.53% Cao, and 0.4% phosphorous. It yields latex containing Caoytchoue, resin, albumin, cerrin, sugar, and malic acid. It is used in Ayurveda for the treatment of diarrhea, dysentery, and piles and has hypoglycemic activity. ,, The extracts of F. benghalensis were also reported to inhibit insulinase activity from liver and kidney. It was also found to inhibit lipid peroxidation. Various extracts of F. benghalensis were screened for its anti-allergic and anti-stress potential in asthma by milk-induced leucocytosis and milk-induced eosinophilia. ,, A decoction of roots of F. bengalensis is used by Ayurveda to boost immune system and to fight number of diseases. Traditionally, the bark and latex are used to treat anemia and gastrointestinal disorders. Latex is aphrodisiac, tonic, maturent, lessen inflammation, useful in piles, nose diseases, gonorrhea, syphilis, billiousenes, dysentery, and inflammation of liver.  The bark extract of F. benghalensis and species of Ficus viz. Ficus racemosa, Ficus religiosa, Ficus inspida, Ficus elastica, Ficus indica, and Ficus carica were found to have anti-inflammatory activity.  However, very little pharmacological investigations of the leaves of F. bengalensis have been conducted. Based on this, an attempt has been made to evaluate inflammatory and analgesic potency of F. benghalensis.
| Materials and Methods|| |
The leaves of the plant F. benghalensis were collected in Chandwad, Nashik, Maharashtra, in October 2008. The plant material was identified and authenticated by the Department of Pharmacognosy, S. S. D. J. College of Pharmacy, Chandwad, Nasik.
Preparation of extract
The leaves were cleaned, dried under shade, and powdered by a mechanical grinder; 400 g of the powder was extracted with solvents in order of their increasing polarity using soxhlet apparatus. The yield of methanolic extract was found to be 5.64%. The methanolic extract of F. benghalensis leaves was administered as a suspension in 2% gum acacia to the animals. From the previous studies in our laboratory, lower doses of the test extract (10, 20, and 100 mg/kg of body weight p.o.) were selected because the effect produced by lower doses were well comparable with the standard drugs.
Freshly prepared F. benghalensis extract was subjected to phytochemical screening tests for the detection of various constituents using conventional protocol. ,
Albino rats of Wistar strain (150-200 g) and Swiss albino mice (25-30 g) of either sex were procured from National Toxicology Centre, Pune. They were housed in standard polypropylene cages and kept under controlled room temperature (24 ± 2°C; relative humidity 60-70%) in a 12-h light-dark cycle. The rats were given a standard laboratory diet and water ad libitum. Food was withdrawn 12 h before and during the experimental hours. All experimental protocols were approved by the Institutional Animal Ethics Committee.
The following chemicals and drugs were used: Carrageenan (Sigma-Aldrich), acetic acid (Hexon Laboratories Ltd., Mumbai), methanol AR (Qualigens, Mumbai), petroleum ether (60-80°C, Qualigens, Mumbai), and ibuprofen (Vikash Pharma, Mumbai) were used during experimental protocol.
Carrageenan-induced hind paw edema
The effect of oral administration of 10, 20, and 100 mg/kg of the methanolic extract of F. benghalensis (MEFB), 20 mg/kg aspirin, or vehicle (saline, 10 ml/kg) on the hind-paw oedema induced by subplantar injection of 0.1 ml carrageenan (1% w/v) was evaluated according to the method of Winter et al.  In short, 0.1 ml of 1% w/v carrageenan was injected into the subplantar tissue of left hind paw of each rat. Swelling of carrageenan-injected foot was measured at 0, 1, 2, and 3 h using Plethysmometer (UGO Basile, Italy). Animals were treated with test extract 1 h before the carrageenan injection. Measurement was carried out immediately before and 3 h following carrageenan injection. Percent inhibition of test drugs was calculated in comparison with vehicle control (100%).
The F. benghalensis leaf extracts (MEFB) were evaluated for analgesic activity in mice using acetic acid-induced writhing and hot plate tests described below.
Acetic acid-induced writhing
The response to intraperitoneal injection of 0.6% acetic acid, i.e., contraction of the abdominal muscle and elongation of the hind limbs, was induced in albino mice of either sex by the method of Koster et al.  and Williamson et al.  In short, animals were pretreated orally with the methanolic extract (10, 20, and 100 mg/kg) and aspirin (20 mg/kg, p.o.). The extract and aspirin were administered 60 min before intraperitoneal injection of 0.6% acetic acid (0.1 ml/10 g), and 10 min later, mice were placed individually in transparent cages, and the number of abdominal writhes was counted over a period of 20 min. The analgesic activity was evaluated in terms of percentage inhibition of writhes.
Hot plate test
The methods of Williamson et al.  and Eddy and Leimback  were used. Mice were placed individually on a thermostatically controlled hot plate, (Orchid scientific, Model-EH-01) maintained at 55°C. The pain threshold is considered to be reached when the animals lift and lick their paws or attempt to jump out of the beaker. The reaction time (jumps, licking one of the hind legs or one of the forelegs, tapping) was measured 0, 15, 30, 60 and 90 min after oral administration of the extract. The animals were first tested for the paw-lick or jump response and only those that reacted after 4 s were used for the experiment. A cutoff time of 60 s was used to avoid harm to the animals.
Results of all the above estimations have been indicated in terms of mean ± SEM. Difference between the groups was statistically determined by analysis of variance (ANOVA) with Dunnett's test multiple comparisons test using GraphPad InStat version 5.00 (GraphPad Software, CA, USA). The level of significance was set at P<0.05.
| Results|| |
Preliminary phytochemical analysis of the extract revealed the presence of alkaloids, flavonoids, glycosides, alkaloids, tannins, and saponins.
Carrageenan-induced hind paw edema
The MEFB (10, 20, and 100 mg/kg, p.o.) significantly (P<0.05) inhibited carrageenan-induced rat paw edema. Maximum inhibition of paw edema was observed in MEFB (100 mg/kg) at 3 h when compared with the control group. Aspirin inhibited carrageenan-induced rat paw edema by 35.21% [Figure 1].
|Figure 1: Effect of varying doses of MEFB on carrageenan-induced rat paw edema. N=5, treatment, mg/kg, data were analyzed using ANOVA and expressed as mean ± SEM followed by Dunnett's test, and differences between means were regarded significant at *P<0.05|
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Acetic acid-induced writhing
The MEFB (10, 20, and 100 mg/kg, p.o.) significantly (P<0.05) reduced the number of abdominal constrictions induced by acetic acid compared with the control group. Maximum inhibition of writhing response by MEFB (100 mg/kg) was 43% comparable to aspirin (20 mg/kg) [Figure 2].
|Figure 2: Effect of varying doses of MEFB on acetic acid induced writhes in mice. N=5, treatment, mg/kg, data were analyzed using ANOVA and expressed as mean ± SEM followed by Dunnett's test, and differences between means were regarded significant at *P<0.05|
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Hot plate test
The MEFB (10 mg/kg) did not show significant (P<0.05) increase in the mean basal reaction time in hot plate method compared with control. The MEFB (10 and 20 mg/kg) showed significant (P<0.05) increase in the mean basal reaction time. The highest nociception inhibition of stimulus exhibited by MEFB (100 mg/kg) was observed at 15 min [Figure 3].
|Figure 3: Effect of varying doses of MEFB on basal reaction time in hot plate method in mice. N=5, treatment, mg/kg, data were analyzed using ANOVA and expressed as mean ± SEM followed by Dunnett's test, and differences between means were regarded significant at *P<0.05|
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| Discussion|| |
The potential of the MEFB for its analgesic and anti-inflammatory effect was investigated. The test extract (MEFB) at the dose of 10, 20, and 100 mg/kg showed significant analgesic and anti-inflammatory activity.
The results of preliminary phytochemical screening of the methanol extract revealed the presence of alkaloids, flavonoids, saponins, and tannins. Analgesic and anti-inflammatory effects of flavonoids, steroids, and tannins have been reported,  hence the analgesic and anti-inflammatory effects produced by the extract may be attributed individually or collectively to the flavonoids and tannins.
The anti-inflammatory and analgesic tests used in this study were chosen in order to test the different nociceptive stimuli, namely cutaneous, chemical visceral stimuli, and thermic. 
The extract caused marked inhibition of carrageenan-induced oedema in rats. Carrageenan-induced inflammation is believed to be biphasic; the early phase (1-2 h) is mainly mediated by histamine, serotonin, and increased synthesis of prostaglandins in the damaged tissue surroundings, and the late phase is sustained by prostaglandins released and mediated by bradykinin, leukotrienes, polymorphonuclear cells, and prostaglandins produced by tissue macrophages.  The inhibitory effect of the extract 100 mg/kg on carageenan-induced inflammation over period of 3 h is similar to the effect of most NSAIDs. This suggests that it acts in later phase probably involving arachidonic acid metabolites that produce edema dependent on neutrophils mobilization. 
Acetic acid-induced abdominal constriction test is used for the evaluation of pheripheral analgesic activity.  The extract showed analgesic activity in acetic acid-induced writhing test in mice. This indicates that the extract possessed peripheral-mediated analgesic activity. The abdominal constriction response is thought to involve in part local peritoneal receptors,  so the extract may have interfered with these peritoneal receptors to bring about analgesia. Acetic acid-induced writhing test has been associated with increase in the levels of prostaglandins E 2 and F 2α in peritoneal fluid  and lipooxygenases,  so the mechanism of activity of the extract may be linked to cyclooxygenases and/or lipooxygenases.
Tail flick and hot plate tests are the most common tests of nociception that are based on a phasic stimulus of high intensity.  Pain induced by thermal stimulus of the hot plate is specific for centrally mediated nociception.  The ability of the extract to prolong the reaction latency to pain thermally induced in mice by the hot plate further suggests central analgesic activity.
Flavonoids isolated from some medicinal plants have been proven to posses antinociceptive and or anti-inflammatory effects,  and it has been shown by Meli et al.  and Dicarlo et al.  that flavonoids also inhibit gastric motility in a dose-dependent, manner. It is therefore possible that the inhibitory effects on anti-nociceptive and anti-inflammatory effects observed in the extract may be attributed in part to its flavonoid content. Flavonoids also inhibit the phosphodiesterases involved in cell activation. Much of this effect is on the biosynthesis of protein cytokines that mediates adhesion of circulating leukocytes to sites of injury. Flavonoids inhibit biosynthesis of prostaglandins, which are involved in various immunologic responses and are the end products of the cyclooxygenase and lipoxygenase parthways.  Protein kinases are another class of regulatory enzymes affected by flavonoids. Inhibition of these enzymes provides the mechanism by which flavonoids inhibit inflammatory processes. 
From the results obtained, F. benghalensis extract does posses significant anti-inflammatory and analgesic activities. The association of both anti-inflammatory and analgesic effects is well documented for various NSAIDS.  The results tend to corroborate the traditional use of F. benghalensis in the treatment of pain and inflammation.
| Conclusion|| |
Taken together, our results suggest the potential of F. benghalensis in treatment of edema and pain and further in-depth research is required for finding the phytocostituent responsible for biological activity of this traditional plant along with its correct mode of action.
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[Figure 1], [Figure 2], [Figure 3]