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 Table of Contents  
ORIGINAL ARTICLE
Year : 2014  |  Volume : 42  |  Issue : 2  |  Page : 74-78

Antipyretic effect of ethanolic extract of Moringa oleifera leaves on albino rats


1 Department of Pharmacology, IMS and SUM Hospital, Bhubaneswar, Odisha, India
2 Department of Anatomy, IMS and SUM Hospital, Bhubaneswar, Odisha, India
3 Department of Pharmacology, SPS, Siksha O Anusandhan (SOA) University, Bhubaneswar, Odisha, India

Date of Submission11-Apr-2013
Date of Acceptance18-May-2014
Date of Web Publication31-Jul-2014

Correspondence Address:
Ayon Bhattacharya
Department of Pharmacology, IMS & SUM Hospital, Kalinga Nagar, B.O. Ghatikia, Bhubaneswar 751003, Odisha
India
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DOI: 10.4103/1110-1415.137810

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  Abstract 

Objective
The aim of the study was to evaluate the antipyretic activity of ethanolic leaf extract of Moringa oleifera using Brewer's yeast-induced pyrexia model.
Materials and methods
It was a randomized controlled experimental study. A total of 60 rats were taken, dividing them in six groups, each containing 10 rats. Ethanolic extract of M. oleifera (EMO) was administered at 50, 100, 200, and 400 mg/kg doses orally to the respective four groups. The control group was fed with normal saline at 2 ml/kg. A 20% suspension of Brewer's yeast in normal saline was injected subcutaneously at a dose of 10 ml/kg body weight below the nape of neck of rats in all groups. Pyrexia developed after 10 h of Brewer's yeast injection and the temperature was recorded. Animals which showed a rise in body temperature to at least 39°C were included in the study, allowing a minimal of six rats in each group, total of 36 rats. Drugs were given after development of pyrexia and temperatures were recorded. Paracetamol at 100 ml/kg orally was taken as the standard drug.
Results
The ethanolic leaf extract of M. oleifera showed significant (P < 0.05) antipyretic activity at 100, 200, and 400 mg/kg. Paracetamol showed significant antipyretic activity from 15 min of drug administration to 12 h. EMO at a dose of 50 mg/kg did not show antipyretic effect. The onset of action of EMO 100 mg/kg was found to be 2 h and that of 200 and 400 mg/kg was found to be 30 min. For all the doses, the antipyretic effect lasted up to 12 h.
Conclusion
The ethanolic leaf extract of M. oleifera exhibited significant (P < 0.05) antipyretic activity at 100, 200, and 400 mg/kg.

Keywords: antipyretic, Brewer′s yeast, Moringa oleifera


How to cite this article:
Bhattacharya A, Behera R, Agrawal D, Sahu PK, Kumar S, Mishra SS. Antipyretic effect of ethanolic extract of Moringa oleifera leaves on albino rats. Tanta Med J 2014;42:74-8

How to cite this URL:
Bhattacharya A, Behera R, Agrawal D, Sahu PK, Kumar S, Mishra SS. Antipyretic effect of ethanolic extract of Moringa oleifera leaves on albino rats. Tanta Med J [serial online] 2014 [cited 2017 Aug 23];42:74-8. Available from: http://www.tdj.eg.net/text.asp?2014/42/2/74/137810


  Introduction Top


Herbal medicines are assuming great importance in the primary healthcare of individual and society. Over 75% of the world population relies on plant extracts for healthcare, and more than 30% of the entire plant species at one time or the other are used for medicinal purposes [1]. The WHO encourages the use of herbal remedies that have proved efficacious in primary healthcare [2].

Pyrexia or fever is defined as the elevation of body temperature. It is a natural defense mechanism or response to tissue damage, inflammation, malignancy, or graft rejection. Because of poor hygiene practices and malnutrition, children from developing countries suffer from various infections, which present as fever. These fevers are accompanied by personal discomforts such as pain (myalgia), which undoubtedly leads to morbidity and mortality [3].

Moringa oleifera, native of the western and sub-Himalayan tracts, India, Pakistan, Asia, Africa, and Arabia, is now cultivated in Philippines, Cambodia, Central America, North and South America, and Caribbean islands [4]. It is a small, fast-growing evergreen or deciduous tree that grows up to 10-12 m in height, with open crown of drooping fragile branches, feathery foliage of trip innate leaves, and thick corky, whitish bark [5]. This plant has been known since the ancient Egyptian and Roman times for the multitude of medicinal and other uses. In the Charaka Samhita, it has been stated to be used for various infections of ear, nose, and throat. M. oleifera is known by various names such as horseradish tree, drumstick tree, ben oil tree, miracle tree, and 'Mother's best friend' [6]. The leaves are of high nutritive value and contain a rare combination of ascorbic acid, carotenes, flavonoids, isoquercetin, and glycosides such as niazirin, 4-hydroxymellein, β-sitosterol, vanillin, caffeoylquinic, kaempferol, and zeatin [7,8]. As a result of the anti-inflammatory action of these bioactive constituents, the antipyretic action can be hypothesized. Antipyretic potential of the ethanolic seed extract of M. oleifera has been studied [9] but not that of leaves. Hence, the present study was undertaken to study the antipyretic potential of leaf extract of M. oleifera.


  Materials and methods Top


The study protocol was approved by Institutional Animal Ethical Committee (IAEC), Siksha O Anusandhan (SOA) University. Experiments were performed in Department of Pharmacology, IMS & SUM Hospital, Bhubaneswar. It was a randomized controlled study.

Collection of plant material

The leaves were collected from the local areas of Syampur (Bhubaneswar, Odisha), and its identity was confirmed taxonomically by Dr P.C. Panda, scientist of Regional Plant Research Centre (RPRC), Bhubaneswar.

Extract preparation

The leaves (500 g) of M. oleifera were obtained locally and shade dried and powdered. It was extracted with 90% ethanol in a Soxhlet apparatus for 8 h. Extract was filtered (Whitman filter paper no. 1), concentrated in a rotary evaporator to yield a semisolid mass of 42 g (8.4% w/w), stored at 4°C, and used for oral administration.

Animals

The animals were procured from central animal house IMS & SUM Hospital. Wistar albino rats of either sex (100-200 g) were used. Food and water were given ad libitum. Animals were acclimatized to laboratory conditions for 7 days before the experiments. The study was approved by the IAEC of SOA University, Bhubaneswar, under the approval number 22/12/IAEC/SPS/SOA. All experiments and animal care were according to the CPCSEA and good laboratory practice guidelines. No animals were killed at the end of the study.

Chemicals

Paracetamol (Dr Reddy's Laboratory, Hyderabad, India), yeast extract powder (HiMedia Laboratories Pvt Ltd, Mumbai, India), and other solvents were of analytical grade.

Brewer's yeast-induced pyrexia

The animals were randomly divided into six groups, each group consisting of 10 rats; a total of 60 rats were used in the study by randomized sampling technique: group I (control, normal saline given orally at 2 ml/kg body weight); group II (standard, paracetamol 100 mg/kg); groups III, IV, V, and VI {ethanolic extract of M. oleifera (EMO) 50, 100, 200, and 400 mg/kg, respectively}. This is regarded as a classical method for antipyresis testing. Wistar strain of albino rats of either sex weighing 100-200 g were used for the study. The animals were fasted for 18 h before commencement of the experiment, but water was provided ad libitum. An initial rectal temperature was recorded using a rectal thermometer to a depth of 1.5 cm in the rectum of rats. Animals with a body temperature between 36 and 38°C were included in the study. A 20% Brewer's yeast in 0.9% w/v saline was injected subcutaneously below the nape of neck at a dose of 10 ml/kg thereafter. The injection site was massaged to ensure the spread of suspension below the skin. Room temperature was maintained at 22-24°C. After the yeast injection, food was immediately withdrawn. After 10 h postchallenge, the rise in rectal temperature was recorded. Animals which showed a rise in body temperature to at least 39°C were included in the study, allowing a minimal of six rats in each group, total of 36 rats. The animals received the standard (paracetamol 100 mg/kg) or the test compound (EMO 50, 100, 200, 400 mg/kg) by oral administration, and the rectal temperature was recorded at 0, 0.25, 0.50, 1, 2, 3, 4, 5, 6, 12, and 24 h after dosing. The maximum reduction in average rectal temperature in comparison with the control hyperpyrexia group was calculated and compared [10].


  Results Top


Subcutaneous injection of the pyrogenic dose of yeast produced elevated changes in rectal temperature, which is shown in [Table 1]. The EMO showed a significant (P < 0.05) decrease in rectal temperature at doses 100, 200, and 400 mg/kg when compared with the control. However, no significant decrease in mean temperature was noted by EMO 50 mg/kg when compared with control throughout the study period ([Table 1] and [Figure 1]. EMO at doses of 50, 100, 200, and 400 mg/kg showed a progressive decline in mean temperature pattern with the increase in the dose [Figure 2]. Paracetamol showed significant (P < 0.05) decrease in rectal temperature from 0.25 to 12 h. The onset of action of paracetamol was 15 min and the body temperature became normal after 12 h. EMO at doses 100 mg/kg showed its onset of action from 2 h and EMO 200 and 400 mg/kg from 30 min [Figure 2]. The line graph [Figure 3] shows that paracetamol registered a phenomenal decrease in mean temperature between 15 min and 2 h, and thereafter maintaining a steady mean temperature. [Figure 4] illustrates that the most significant (P < 0.05) decrease in round-the-clock mean temperature in this study was shown by paracetamol followed by EMO at 400 mg/kg. Statistical analysis was performed using one-way analysis of variance followed by post-hoc Dunnette's test. The results were recorded as mean ± SD.
Table 1 Effect of paracetamol and ethanolic extract of Moringa oleifera on yeast-induced pyrexia

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Figure 1: Line graph showing the effect of paracetamol and ethanolic extract of Moringa oleifera (EMO) on yeast-induced pyrexia.

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Figure 2: Line graph showing the effect of ethanolic extract of Moringa oleifera (EMO) 100, 200, and 400 mg/kg in comparison with control on yeastinduced pyrexia model. *P < 0.05, **P < 0.01, ***P < 0.001.

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Figure 3: Line graph showing the effect of ethanolic extract of Moringa oleifera (EMO) 400 mg/kg and paracetamol on yeast-induced pyrexia. *P < 0.05, **P < 0.01, ***P < 0.001.

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Figure 4: Radar diagram showing the effect of ethanolic extract of Moringa oleifera (EMO) and paracetamol on yeast-induced pyrexia model.

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  Discussion Top


Brewer's yeast (lipopolysaccharide, which is the cell wall component of Gram negative bacteria) is an exogenous pyrogen that binds to the immunological protein called the lipopolysaccharide binding protein [11]. This binding results in the synthesis and release of various endogenous cytokine factors such as interleukin (IL)-1, IL-6, and TNFa, which activate the arachidonic acid pathway, and ultimately result in the synthesis and release of prostaglandin E 2 (PGE 2 ) [12]. Yeast-induced pyrexia is called as pathogenic fever [13].

According to the classical view, fever is induced by inflammatory mediators (IL-1, IL-2, TNFα, others) released by the peripheral mononuclear macrophages and other immune cells [14,15]. These fever-promoting cytokines are transported from blood to the brain by specific carriers [16]. Cytokines are transported by bloodstream and enter the brain through the circumventricular organs [17]. Alternatively, the cytokines could interact with their receptors on brain endothelial cells [18] or perivascular tissue [19]. This assumed mechanism of fever induction is known as the humoral hypothesis of fever induction. These proinflammatory mediators act on the preoptic/anterior hypothalamus triggering the release of PGE 2 produced from cyclooxygenase (COX)-2, and thus elevating the body temperature [20].

An effective febrifuge such as paracetamol acts by blocking the effect of these pyrogens in the temperature-sensitive neurons in the preoptic region of the hypothalamus to COX formation of PGE 2 [21]. However, in inflammatory lesions, paracetamol is a poor inhibitor of COX due to presence of peroxides.

The anti-inflammatory activity of M. oleifera is attributed to its COX inhibitory activity [22]. Thus, the present study postulates that EMO could reduce pyrexia by reducing the concentration of PGE 2 in the hypothalamus or by interrupting the steps that connect the peripheral inflammation with the central production of PGE 2 or both [23,24].

The phytochemical ingredients in the leaf extract, such as phenolics, flavonoids, tannins, saponins, terpenoids, isoquercetin, and glycosides such as niazirin, 4-hydroxymellein, β-sitosterol, and vanillin, could be responsible for its antipyretic activity [6,25].


  Conclusion Top


Therefore, from the results of the study, we can deduce that EMO could be used as an antipyretic agent at 100, 200, and 400 mg/kg. However, further studies are necessary to examine the mechanism of action of the antipyretic activity and to isolate the active compounds responsible for this pharmacological activity.


  Acknowledgements Top


 
  References Top

1.Joy PP, Thomas J, Mathew S, et al. Medicinal plants. Tropical horticulture (vol. 2). Calcutta: Naya Prokash; 2001. 449-632.  Back to cited text no. 1
    
2.Amos S, Kolawole E, Akah P, et al. Behavioural effects of the aqueous extract of Guiera senegalensis in mice and rats. Phytomedicine 2001; 8:356-361.  Back to cited text no. 2
    
3.Ighodara I, Raymond IO, Steve OO, et al. Antipyretic and analgesic effects of the aqueous extract of the fruit pulp of Hunteria umbellata K schum (Apocynaceae). Trop J Pharm Res 2009; 8:331-336.  Back to cited text no. 3
    
4.Durgesh KD, Jyotsna D, Anil K, et al. A multipurpose tree - Moringa oleifera. Int J Pharm Chem Sci 2013; 2:415-423.  Back to cited text no. 4
    
5.Roloff A, Weisgerber H, Lang U, et al. Moringa oleifera Lam 1785 (ISBN: 978-3-527-32141-4). Enzyklopädie der Holzgewächse, Handbuch und Atlas der Dendrologie. WILEY-VCH Verlag GmbH & Co Weinheim, 2009.  Back to cited text no. 5
    
6.Vinoth B, Manivasagaperumal R, Balamurugan S. Phytochemical analysis and antibacterial activity of Moringa oleifera lam. Int J Res Biol Sci 2012; 2:98-102.  Back to cited text no. 6
    
7.Goyal BR, Agrawal BB, Goyal RK, Mehta AA. Phyto-pharmacology of Moringa oleifera Lam.: an overview. Nat Prod Radiance 2007; 6:347-353.  Back to cited text no. 7
    
8.Saluja MP, Kapil RS, Popli SP. Studies in medicinal plants: Part VI. Chemical constituents of Moringa oleifera Lamk. (hybrid variety) and isolation of 4-hydroxymellein. Indian J Chem B 1978; 16:1044-1045.  Back to cited text no. 8
    
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11.Ukwuani AN, Abubakar MG, Warra SH, et al. Antipyretic activity of some Nigerian medicinal plants in rats. Int J Pharm Res 2012; 4:48-51.  Back to cited text no. 11
    
12.Gege-Adebayo GI, Bassi AS, Igbokwe VU, et al. Antipyretic effect of Ocimum gratissium on Brewer's yeast induced fever in Wistar rats. J Med Med Sci 2013; 4:247-251.  Back to cited text no. 12
    
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14.Zeisberger E. From humoral fever to neuroimmunological control of fever. J Therm Biol 1999; 24:287-326.  Back to cited text no. 14
    
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16.Banks WA, Plotkin SR, Kastin AJ. Permeability of blood brain barrier to soluble cytokine receptors. Neuroimmunomodulation 1995; 2:161-165.  Back to cited text no. 16
    
17.Roth J, Harre EM, Rummel C, et al. Signaling the brain in systemic inflammation: role of sensory circumventricular organs. Front Biosci 2004; 9:290-300.  Back to cited text no. 17
    
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21.Ashok BK, Ravishankar B, Prajapati PK, et al. Antipyretic activity of Guduchi Ghrita formulations in Albino Rats. Ayu 2010; 31:367-370.  Back to cited text no. 21
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23.Li S, Dou W, Tang Y, et al. Acetaminophen: antipyretic or hypothermic in mice? In either case, PGHS-1b (COX-3) is irrelevant. Prostaglandins Other Lipid Mediat 2008; 85:89-99.  Back to cited text no. 23
    
24.Eun-Jung P, Sarot C, Leng Chee C, et al. Inhibition of lipopolysaccharide-induced cyclooxygenase-2 expression and inducible nitric oxide synthase by 4-[(2¢-O-acetyl-α-l-rhamnosyloxy)benzyl]isothiocyanate from Moringa oleifera. Nut cancer 2011; 63:971-982.  Back to cited text no. 24
    
25.Golam AM, Shibly NM, Mamun AA. Phytochemical screening and antipyretic effect of Curcuma zedoaria Rosc. (Zingiberaceae) rhizome. Br J Pharm Res 2014; 4:569-575.  Back to cited text no. 25
    


    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4]
 
 
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