Research articles
 

By Mr. Virendra Yadav , Dr. Soma Jayalakshmi , Mr. Rajeev K Singla , Dr. Arjun Patra , Dr. Salim Khan
Corresponding Author Mr. Rajeev K Singla
Sadbhavna College of Management & Technology, Raikot - India 124001
Submitting Author Mr. Rajeev K Singla
Other Authors Mr. Virendra Yadav
College of Pharmacy, Institute of Foreign Trade & Management, Lodhipur rajput, Moradabad, Uttarprade, - India

Dr. Soma Jayalakshmi
College of Pharmacy, Institute of Foreign Trade & Management, Lodhipur rajput, Moradabad, Uttarprade, - India

Dr. Arjun Patra
College of Pharmacy, Institute of Foreign Trade & Management, Lodhipur rajput, Moradabad, Uttarprade, - India

Dr. Salim Khan
Department of Botany & Microbiology, College of Science, King Saud University, Riyadh-11451, - Saudi Arabia

PHARMACOLOGY

Callicarpa macrophylla; Analgesic; Anti-inflammatory; Roots.

Yadav V, Jayalakshmi S, Singla RK, Patra A, Khan S. Assessment of Anti-Inflammatory and Analgesic Activities of Callicarpa Macrophylla Vahl. Roots Extracts. WebmedCentral PHARMACOLOGY 2012;3(5):WMC003366
doi: 10.9754/journal.wmc.2012.003366

This is an open-access article distributed under the terms of the Creative Commons Attribution License(CC-BY), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
No
Submitted on: 13 May 2012 04:41:18 PM GMT
Published on: 14 May 2012 12:20:35 PM GMT

Abstract


Callicarpa macrophylla, an indigenous plant of India, had been the plant of study for the current research work. Aqueous as well as ethanolic extracts of its roots(at two concentrations 200 & 400 mg/kg) were evaluated for its analgesic and anti-inflammatory potentials using tail immersion test and carrageenan paw edema method in albino rats respectively. Aqueous extract of roots are having better analgesic activity than that of its ethanolic extract. Whereas ethanolic root extract have superior anti-inflammatory spectrum than aqueous one. Results are highly promising and ascertain that roots of C. macrophylla have analgesic and anti-inflammatory potential, comparable to that of standards.

Introduction


The Process of drug discoveries/inventions is elaborate, requiring, on an average 8 to 10 years  and costing millions to reach a new drug to the market[1]. At present, approximately 25% of drugs in modern pharmacopoeia were derived from plants(phytomedicines) and many others were synthetic analogues built on the prototype compounds isolated from plants. Indian folk medicine comprises of numerous prescriptions for therapeutic purposes such a healing of wounds, inflammation, skin infections, leprosy, diarrhea, scabies, venereal diseases, ulcers, snake bite etc[2-4].

Callicarpa macrophylla Vahl. (fam-Verbenaceae) is an erect shrub which is globally distributed across India, Nepal, Bhutan, Myanmar, South East Asia, and China[5]. Medicinal Plants are capable of synthesizing an overwhelming variety of low – molecular weight organic compounds called secondary metabolites, usually with unique  and complex structures[6]. Interest in the roots of this plant C. macrophylla has been heightened by reports of its traditional uses as anti-pyretic, analgesic, anti-ulcer, gastric stimulants etc[7-9].

Keeping this views in mind, aim of current research work was to evaluate analgesic and anti-inflammatory potential of C. macrophylla roots.

Materials and Methods


Collection & Authentification of Plant Material

The drugs were collected from Banaras Hindu University campus, Varanasi and authenticated by Dr. V.K. Joshi, Dean of Faculty of Ayurveda, Institute of Medical Science, B.H.U., Varanasi and also through National Botanical Research Institute (NBRI), Lucknow. A Voucher specimen of all the plants has been preserved in the Department of Pharmacognosy, College of Pharmacy, IFTM, Moradabad, for further references. The collected leaves were shade dried 15 days and size reduced by laboratory grinder in to coarse powder. The air dried coarse powder is used for preparation of extract.

Preparation of Extracts

The ethanolic and aqueous extracts were prepared according to the standard procedure[10]. The filtered, extracts were dried in a vacuum evaporator and aqueous & alcoholic extracts were kept in desiccators until further use.

Animals

Male / female albino rats weighing between 120 to150 grams, from Animal House, College of Pharmacy, IFTM, Moradabad, were divided in ten groups of six animals each. The animals were kept in polypropylene cages, under standard condition of 12:12 light and dark cycle.

Evaluation of Analgesic Activity using Tail Immersion Model

     Rats (six per group) were used. Rats was administered orally with vehicle (3ml/kg), pentazocine (30mg/kg), ethanolic, aqueous extract (200 and 400 mg/kg) of roots. The distal part of tails (3c.m) of the animals was immersed in hot water at a temperature of 55±0.5°C. The time taken to withdraw the tail was noted as reaction time with a stopwatch. A cut off time of 10 sec was maintained at55 ±0.5°C ° to prevent tissue damage. The reaction time was measured at 0, 15, 30, 45 and 60 min after treatment, respectively.

Evaluation of Anti-inflammatory Activity Using Carrageenan Paw Edema Method

 Experimentally inflammation was produced by carrageenan paw edema method in albino rats[5,11]. A volume of 0.01 ml of 1% (w/v) carrageenan suspension in (0.9 % w/v sodium chloride) was injected through a 26-gauge needle into the plantar side of the left hind paw. The prepared drug samples 200 mg/kg, 400 mg/kg of the ethanolic and aqueous extract orally were administered one hour before the carrageenan injection. The standard drug Diclofenac sodium was given orally in dose of 20mg/kg. Tween 80 (1% v/v) was used as suspending agent. The volume of the paw was measured at 60, 120, 180 and 240 minutes after injection. The ankle joint of the rats was marked with permanent marker and the paw was dipped in the mercury. The volume of hind paw of the rats up to the ankle joint was measured plethysmographically by the mercury displace method. The percent inhibition was calculated by following formula-                 

% Inhibition =   (1-Vt / Vc) x 100

Where, Vt and Vc are the mean change in paw volume of treated and control rats respectively

Data analysis and statistics

The values were expressed as mean ± standard error mean (SEM). Statistical analysis of the data was carried out by two way ANOVA followed by bonferroni test to determine the significant between two groups p<0.05 was considered significant.

Results and Discussion


A significant reduction of the painful sensation due to tail immersion in warm water was observed followed oral administration of the ethanolic and aqueous extract at dose of 200, 400mg/kg of leaves and roots of C. macrophylla Vahl. The effect was found to the dose dependent. In this model, higher dose of the aqueous extract (400mg/kg) at an interval of 60 min has exhibited better analgesic activity than the standard drug. Representation of result of analgesic activity was shown in Table 1 & Figure 1.

Several flavonoids isolated from medicinal plant have been discovered to posses significant analgesic effects (Gulnur et al., 2004 ;). The analgesic activity of ethanolic, aqueous extract of roots of C. macrophylla Vahl. may be due to the presence of flavonoids compound.

Table 1 Effect of ethanolic, aqueous extract of C. macrophylla Vahl. roots on pain using tail immersion test in rats

 

Treatment

 

Dose

mg/kg

 

Average tail withdrawing time (s)

0 min

15 min

30 min

45 min

60 min

Control

      -

4.11 ± 0.20

4.25 ± 0.19

4.20 ± 0.22

4.32 ± 0.09

4.28 ± 0.18

Pentazocine

30

4.52 ± 0.20

6.66 ± 0.28*

8.63 ± 0.16*

8.72 ± 0.48*

9.48 ± 0.19*

REE

200

4.20 ± 0.39

4.32 ± 0.30

5.85 ± 0.41*

6.02 ± 0.09*

6.96 ± 0.35*

400

4.17 ± 0.31

4.69 ± 0.18

6.79 ± 0.26*

7.60 ± 0.19*

8.72 ± 0.15*

RAE

200

4.02 ± 0.26

4.36 ± 0.31

5.96 ± 0.21*

8.20 ± 0.18*

8.86 ± 0.22*

400

3.99 ± 0.20

5.62 ± 0.32*

6.79 ± 0.19*

9.50 ± 0.35*

9.96 ± 0.21*

 

 

 

Treatment

Dose

(mg/kg)

Increase in paw volume (in ml) after different times

     %Reduction

 (after 4 hr)

30 min

1hr

2hr

3hr

4hr

Control

-

 0.28  ± 0.02

 0.36 ± 0.01

 0.69 ± 0.03

 0.73 ± 0.01

 0.72 ± 0.01

 

   Diclofenac    

  Sodium

20

 0.18  ± 0.01*

 0.21 ± 0.01*

 0.23 ± 0.02*

 0.21 ± 0.01*

 0.16 ± 0.03*

       77.78

   REE

200

 0.22 ± 0.03*

 0.24 ± 0.02*

 0.37 ± 0.01*

 0.32 ± 0.04*

 0.30 ± 0.01*

        58.33

400

 0.20 ± 0.01*

 0.23 ± 0.01*

 0.32 ± 0.04*

 0.28 ± 0.01*

 0.18 ± 0.04*

        75.00

    RAE

     200

 0.23 ± 0.02

 0.24 ± 0.01*

 0.38 ± 0.01*

 0.34 ± 0.03*

 0.32 ± 0.01*

        55.56

     400

 0.21 ± 0.01*

 0.24 ± 0.02*

 0.33 ± 0.02*

 0.28 ± 0.01*

 0.22 ± 0.02*

         69.44

Table 2 Effect of ethanolic, aqueous extract of C. macrophylla Vahl. leaves and roots on Carrageenan induced paw oedema in rats

 

Carrageenan has been widely used as a noxious agent able to induce experimental inflammation for the screening of compounds possessing anti-inflammatory activity. This phlogistic agent, when injected locally into the rat paw, produced a severe inflammatory reaction, which was discernible within 30 min. The development of edema induced by carrageenan corresponds to the events in the acute phase of inflammation, mediated by histamine, bradykinin and prostaglandins produced under an effect of cyclooxygenase. (Borgi et al., 2007).

 The ethanolic and aqueous extracts (200 mg/kg, 400 mg /kg) of root of Callicarpa macrophylla showed significant (p< 0.05)  anti-inflammatory effect in the acute phase of the inflammation process as compared with standard drug, Diclofenac sodium (20 mg/kg) body wt. as shown in Table 2 & Figure 2. Further, the ethanolic and aqueous extracts were found to contain carbohydrates, steroids, flavonoids and tannins, through preliminary photochemical screening. The anti-inflammatory activity may be due to one/more group of above Phytoconstituents which may cause inhibition of histamine, serotonin or prostaglandin synthesis.

 

Acknowledgment

Authors would like to express their gratitude towards the management of IFTM & Head, College of Pharmacy for providing financial support to execute this research plan.

Declaration of Funding and Competing Interests

This research was solely funded by Management, IFTM, U.P, India. Authors declare no competing interests.

References


1. Manthan D Janodia et al. Patents, Health Policy and Access to Medicines. Indo Global Journal of Pharmaceutical Sciences. 2011; 1(1): 33-38.
2. C C Barua et al. Impact of Achyranthes aspera L. on Protein Profile  in Impaired Wound Models. Indo Global Journal of Pharmaceutical Sciences. 2011; 1(1): 13-24.  
3. Sofowora, A., Medicinal Plants and Traditional Medicine. In: Africa, John Willey & Sons Ltd. New York, 1982, pp. 6.
4. Biswas, T. K., Maity, L. N.  and  Mukherjee, B., Wound healing potential of Pterocarpus santalinus Linn: a pharmacological evaluation. Int. J. Low Ext. Wounds., 2004, 3, 143–150.
5. Virendra Yadav, Soma Jayalakshmi, Rajeev K Singla, Arjun Patra. Preliminary Assessment of Anti-inflammatory Activity of Callicarpa macrophylla Vahl. Leaves Extracts. Indo Global Journal of Pharmaceutical Sciences. 2011; 1(3):219-222.
6. Salim Khan, Rajeev K Singla, Malik Zainul Abdin. Assessment of Phytochemical Diversity in Phyllanthus amarus using HPTLC Fingerprints. Indo Global Journal of Pharmaceutical Sciences. 2011; 1(1): 1-12.
7. Shome, Usha. Rawat, A.K.S and Mehrotra, Shanta. Comparative Pharmacognosy of two Callicarpa Spp, and Commercial Sample of Priyangu. Ethnobotany. 1997; 9: 24-34.
8. Jain, S.P.  Puri, H.S. Ethanomedicinal Plants of Jounsar Bawar Hills, Uttar Pradesh India. Journal of Ethnopharmacology. 1984; 12: 213-222.
9. Anonymous. The Useful plant of India, National Institute of Science Communication, CSIR, New Delhi. 2000, pp. 96.
10.  Girish R Bankar et al. Vaosrelaxant & Antihypertensive Effect of Cocos nucifera Linn. Endocarp on isolated rat thoracic aorta and DOCA salt induced hypertensive rats. Journal of Ethnopharmacology. 2011; 134: 50-54.
11. Vogel, H. Gerhard. Drug Discovery & Evaluation, Pharmacological Assay, 2002, 2nd Edition, pp. 696, 697, 759-760, 772.

Source(s) of Funding


None

Competing Interests


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