Evaluation of analgesic potential of Boerhavia diffusa roots in albino mice

 

Kadiri Sunil Kumar¹*, V. Saideepthi², Shaik Shahajeb², M. Nandini², R.Suthakaran³

¹Associate Professor, Department of Pharmacology, Vijaya College of Pharmacy, Munaganoor – 501511, Hyderabad, Telangana, India

²B-Pharmacy Final year II Sem, Vijaya College of Pharmacy, Munaganoor – 501511,

Hyderabad,  Telangana, India

³Principal and Professor, Department of Pharmaceutical Chemistry, Vijaya College of Pharmacy,

Munaganoor – 501511, Hyderabad, Telangana, India

 

ABSTRACT:

The present research was designed to evaluate the analgesic potential of Boerhavia diffusa root powder 150 mg/kg, p.o in albino mice. Boerhavia diffusa is a popular herb in ayurveda which is also known as Punarnava. Boerhavia diffusa root powder was procured from a local ayurvedic store. The analgesic effect of the root powder was evaluated by using acetic acid induced writhings test in mice. It was found that Boerhavia diffusa root powder at a dose of 150 mg/kg treated for 15 days has significantly inhibited pain induced by acetic acid by decreasing the number of writhing responses such as extension of hind limbs (22±0.77), abdominal constrictions (13±0.08) and trunk twisting (20±0.55) when compared to control treated mice i.e extension of hind limbs (38±0.17), abdominal constrictions (26±1.23) and trunk twisting (24±0.55) in control mice. The results were compared with standard diclofenac sodium (25 mg/kg, p.o) treated mice. Hence it is evident from the above results that boerhavia diffusa at 150 mg/kg possesses significant analgesic potential.

 

 

 

INTRODUCTION:

Allopathic medicines play a predominant role in the modern society [1]. There are several potent pain killers available in the modern medicines like NSAIDS and opioids which can quickly minimize the intensity of pain but may simultaneously lead to severe adverse effects like GI disorders, hepatotoxicity, nephrotoxicity, tolerance, dependence and addiction [2], [3]. The search for alternatives to mitigate pain in allopathy has remained crucial [4].

 

Treatment of pain with herbal medicines could be a better option to get rid of the side effects caused by allopathic pain killers [5]. Boerhavia diffusa is a flowering plant that is commonly known as punarnava which means rejuvenating or renewing the body [6]. The plant has very rich medicinal properties. Punarnava cures digestive problems, alleviates fluid retention, useful in treatment of heart diseases, beneficial in anaemia, hernia, respiratory problems, and liver disorders and manages to maintain the cholesterol and lipid levels in normal healthy limits [7]. The plant is rich in sitosterol, esters of sitosterol, alkaloids, arachidic acid, boerhavic acid, flavonoids etc [8]. Literature survey reveals that flavonoids have significant analgesic activity [9]. Phytochemical screening of punarnava roots indicates the presence of significant amounts of flavonoids. Hence the aim of our research is to evaluate analgesic potential of punarnava root powder by acetic acid induced writhings test in albino mice. Till today no one has reported the analgesic activity of Boerhavia diffusa roots. Therefore the current research is essential and justifiable.

 

MATERIALS AND METHODS:

Fresh root powder of Boerhavia diffusa was procured from balu herbals, champapet, Hyderabad. The root powder of Boerhavia diffusa was authenicated by Dr. N. Sivaraj, eco-botanist, national bureau of plant genetic resources, rajendranagar, hyderabad. Appropriate concentration of stock solution of root powder and standard diclofenac sodium was prepared by using 2% acacia suspension and used for analgesic activity. Acetic acid 1% was prepared by using distilled water.

 

Preliminary Phytochemical Screening [10]:

Preliminary phytochemical tests were performed for the root powder of Boerhavia diffusa to detect the presence of phytoconstituents by following the standard methods described in the practical pharmacognosy of kokate and khandelwal. The results have been tabulated in table I.

 

Experimental Animals:

Albino mice (20-25g) were used for the present analgesic activity. They were procured from sainath agencies, musheerabad (282/99/CPCSEA). After randomization into various groups and before initiation of experiment, the rats were acclimatized for a period of 10 days. Animals were housed in polypropylene cages and maintained under standard environmental conditions such as temperature (26 ± 2ºc), relative humidity (45-55%) and 12hr dark/light cycle. The animals were fed with rodent pellet diet (Golden Mohur Lipton India Ltd.) and water ad libitum. The study protocol was approved from the institutional animal ethics committee (IAEC) before commencement of experiment IAEC (1292/ac/09/CPCSEA).

 

Determination of Acute Toxicity:

The safer therapeutic dose of Boerhavia diffusa i.e 150 mg/kg in mice was selected from literature by referring to various research articles of boerhavia diffusa. [11]

 

Effect of Boerhavia diffusa root powder on acetic acid induced pain in albino mice:

The experiment was performed on albino mice (20-25gms) of either sex procured from sainath agencies, musheerabad. The animals were housed in colony cages at an ambient temperature of 26±2⁰C and relative humidity (45-55%) with a 12h/12h light dark cycle and access to food and water ad libitum. Food was restricted during the experiment. Stock solutions of Boerhavia diffusa (150 mg/kg) and diclofenac sodium (25 mg/kg) were prepared in 2% acacia suspension. Acetic acid (1%) was prepared in distilled water. Divide the animals in to three groups control(C), test (T), and standard(S) each group consisting of six mice. The control, test and standard group mice were treated with distilled water, p.o, Boerhavia diffusa (150 mg/kg, p.o) root powder, and standard diclofenac sodium (25 mg/kg, p.o) for 15 days. On 15^th day the analgesic activity was determined by inducing pain with intraperitoneal injection of 1% acetic acid one hour after administration of distilled water, Boerhavia diffusa (150 mg/kg), root powder and standard diclofenac sodium to control, test and standard groups and the writhing responses to pain such as number of abdominal contractions, trunk twisting and extension of hind limbs were recorded for a period of 10 minutes. The mean writhing scores in control, Boerhavia diffusa and diclofenac sodium and treated groups were calculated. The inhibition of pain response by Boerhavia diffusa was noted and compared with the control and standard treated mice.

 

Figure I: Extension of hind limbs

 

Figure II: Trunk twisting

 

Figure III: Abdominal Constriction

 

Figure I, II and III: Writhing responses seen in mice after intraperitoneal injection of 1% acetic acid

 

 

STATISTICAL ANALYSIS:

The values are represented as mean ± S.E.M and statistical significance between treated and control groups was analyzed using One way ANOVA, followed by Dunnett’s test where ** P<0.01 and *P<0.05 was considered statistically significant.

 

RESULTS AND DISCUSSION:

Boerhavia diffusa (Punarnava) is a well known herb in ayurvedic community due to its wide variety of applications in the treatment of various disorders. Phytochemical analysis of root powder of Boerhavia diffusa reveals the presence of sitosterol, esters of sitosterol, alkaloids, arachidic acid, boerhavic acid and flavonoids. It is evident from the literature that flavonoids have potent analgesic activity. In acetic acid induced writhing test in mice it was found that after treatment with Boerhavia diffusa for 15 days the number of writhing responses has significantly decreased in test group mice when compared to control group. The results are tabulated in table no.II. As the writhing responses are the index of pain produced in mice, reduction in writhing responses with Boerhavia diffusa root powder at 150 mg/kg indicates that there is a pain inhibiting potential with Boerhavia diffusa.

 

Table I: Preliminary Phytochemical screening of chloroform extract of Boerhavia diffusa leaves.

Phytoconstituents	Boerhavia diffusa root powder
Carbohydrates Steroids Glycosides Flavonoids Alkaloids Tannins proteins Triterpeniods Resins Phytosterols Gum and mucilages	-   +   +   +   +   -   -   +   -   +   -

-  Absent   +   Present

 

Table II: Analgesic effect of Boerhavia diffusa root powder in acetic acid induced writhing responses in mice

Sl.No	Group	Writhing Responses
		EHL	AC	TT
1	Control (Distilled water)	38 ± 0.17	26 ± 1.23	24± 0.55
2	Test (Boerhavia diffusa root powder, 150 mg/kg, p.o)	22 ± 0.77*	13 ± 0.08*	20± 1.79
3	Standard (diclofenac sodium, 25 mg/kg, p.o)	19 ± 0.18**	12 ± 0.08*	13± 0.16*

P<0.01** and P<0.05* was considered statistically significant.

EHL= Extension of hind limbs, AC= abdominal constriction, TT= trunk twisting

 

Figure IV: Graphical representation of analgesic effect of Boerhavia diffusa root powder in albino mice by acetic acid induced writhing method.

 

 

CONCLUSION:

The present research shows that Boerhavia diffusa at a dose of 150 mg/kg significantly inhibited pain induced by acetic acid in mice which can be seen by reduction in writhing responses in test group. Hence we conclude that the roots of Boerhavia diffusia possess significant analgesic potential. However further studies has to be done to find out the exact active constituent responsible for analgesic activity with its mechanism involved in pain reduction.

 

ACKNOWLEDGEMENTS:

The authors are grateful to the management of Vijaya College of Pharmacy, Hyderabad for providing the facilities for our Research.

 

REFERENCES:

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Received on 05.04.2017          Modified on 28.04.2017

Accepted on 09.05.2017      ©AandV Publications All right reserved