Shehu et al, Afr. J. Pharmacol. Ther. 2016.

5(2): 65-73

African Journal of Pharmacology and Therapeutics Vol. 5 No. 2 Pages 65-73, 2016
Open Access to full text available at http://journals.uonbi.ac.ke/ajpt

Research Article

Acute Toxicological, Analgesic and Anti-

Inflammatory Effects of Methanol Extract of
Laggera aurita Linn F (Compositae) in Mice and
Rats
Aishatu Shehu a,*, Temidayo O. Olurishe a, Abdulkadir U. Zezi a, and Abubakar Ahmed b
a Department of Pharmacology and Therapeutics, Ahmadu Bello University, Nigeria
b Department of Pharmacognosy and Drug Development, Ahmadu Bello University, Nigeria
_____________
* Corresponding author: Department of Pharmacology & Therapeutics, Ahmadu Bello University, PMB 1045, Zaria-
Nigeria; Tel: +234-803-6028298; Email: [email protected]

Background: The plant Laggera aurita is an annual or biannual plant belonging to the family Compositae that has
been used for management of pain related conditions locally. It reportedly has anti-oxidant as well as antimicrobial
properties.
Objectives: To conduct LD50 and phytochemical studies, evaluate the analgesic and anti-inflammatory properties of
the methanol extract of L. aurita and determine possible mechanism of action.
Methodology: Analgesic and anti-inflammatory properties of the extract were investigated using acetic acid induced
writhing, thermally-induced pain, and formalin induced inflammation in rats and mice. Phytochemical and acute
toxicological screenings were also conducted.
Results: The LD50 was found to be above 5000 mg/kg with slight changes in histological architecture observed in the
kidney, liver, lungs and stomach. The extract at dose 200, 400 and 800 mg/kg significantly (p<0.05) inhibited acetic
acid induced writhes in mice and increased mean reaction time in the thermal pain model, both dose dependently. The
effect on thermally induced pain was blocked by naloxone, a non-specific opioid antagonist, suggesting opioid
receptor involvement in analgesia. The extract also significantly (p<0.05) decreased formalin induced paw edema
dose dependently.
Conclusion: These findings suggest that the methanol extract of L. aurita possesses analgesic and anti-inflammatory
properties that justify its ethnomedicinal use in management of pain and inflammation.
Keywords: Laggera aurita, anti-inflammatory, analgesic, acute toxicity.

Received: January, 2016

Published: May, 2016

1. Introduction Abanaadene (vulture’s excrement) in Igbo (Awka) and

Eru-taba (Slave of tobacco) in Yoruba (Ilorin) (Burkill,
The use of herbal drugs in the treatment of pain is a 1985). L. aurita is a widely used medicinal plant in
common practice in many African countries (Usman et African countries like Nigeria, Senegal and Ghana. The
al. 2008). Despite the immense technological plant was also listed among anti tubercular plants of
advancement in modern medicine, many people in foreign origin (Vikrant, 2011). The anti-oxidant and
developing countries still rely on traditional medicine antibacterial activities of the steam distilled volatile oil
and healing practices for their daily health care needs of L. aurita grown in Pakistan have been reported
(Gedif and Hahn, 2003). The Plant L. aurita (Linn F) (Shahwar et al. 2012). Recent studies on the ethanol
belongs to the family Compositae. It is called crude extract of L. aurita showed the presence of

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 Shehu et al, Afr. J. Pharmacol. Ther. 2016. 5(2): 65-73

triterpenes, saponins, flavonoids, coumarins, tannins groups each of three animals were fasted prior to
and also anti-inflammatory effects (Abdulla et al. 2013). dosing (food but not water was withheld overnight for
Olurishe and Mati, (2014) reported some preliminary the rats and for 3 hours for mice). The fasted body
antihyperalgesic potentials. The current study was weight was determined for each animal and the dose
conducted to investigate the acute toxicological and was then calculated according to the body weight. Food
anti-nociceptive effect of the methanol extract of L. was further withheld for 3-4 hours in rats and 1-2 hours
aurita using rodent models. in mice after the extract was administered. The extract
was administered in a single oral dose using an oral
2. Materials and Methods canula. A start dose of 2000 mg/kg was used for each
animal in the first phase. Animals dosed in the first
2.1 Plant Collection phase were observed for 48 hours after which there
was no death and the test proceeded to the second
The whole plant was collected from Zaria, Kaduna State phase. The same procedure was adopted but with a
in December, 2012. The plant specimen was identified dose of 5000 mg/kg.
at the Herbarium Unit, Department of Biological
Sciences, Ahmadu Bello University Zaria- Nigeria by Animals were observed individually at least once during
comparing with a specimen voucher number 510 the first 30 minutes after dosing and periodically during
previously deposited in the herbarium. the first 24 hours with careful observation during first 4
hours, and then daily for 14 days. Observations included
2.2 Extraction Procedure changes in skin and fur, eyes and mucous membranes,
somator activity and behaviour pattern, autonomic and
The plant material was dried under shade with central nervous systems. Animals were also observed
intermittent weighing until a constant weight was for tremors, convulsions, salivation, diarrhoea, lethargy,
obtained. The stalk part of the plant was removed and sleep and coma, with onset of toxic symptoms and
whole plant was size-reduced using mortar and pestle. disappearance also noted. Individual weights of animals
About 1500 grams of the powdered materials was were determined at least weekly. At the end of the test,
extracted with methanol using soxhlet apparatus at a surviving animals were weighed and then euthanized
temperature of 65 °C. The extract was concentrated using chloroform. The liver, kidney, spleen, lungs, heart
under reduced pressure and temperature 45 °C. The and stomach of the animals were harvested fresh and
extract was then stored in a dessicator until required fixed in 10% v/v formalin for histological observations.
for each study.
2.7 Histological studies
2.3 Animals
Tissues were prepared and slides were examined
Wistar strain rats (150-200 grams) and swiss albino microscopically for pathological lesions as described by
mice (20-25 grams) of either sex were obtained from Arthur and John (1978).
the Animal House Facility of the Department of
Pharmacology and Therapeutics, Ahmadu Bello 2.8 Antinociceptive Studies
University Zaria, Nigeria. They were housed in standard
propylene cages and kept under natural day and light Acetic acid induced writhing test
cycle. The animals were fed on standard laboratory
animal diet and water ad libitum. Food was withdrawn The acetic acid induced writhing test in mice as
during the experimental hours to verify described by Koster et al. (1959) was employed. Thirty
pharmacological outcomes. All experimental protocols albino mice were divided into five groups of six mice.
were approved by the University Animal Ethics The first group received 10 ml/kg of normal saline
Committee. orally and this served as control. Group 2 received
Piroxicam 20 mg/kg as positive control. Group 3, 4 and
2.4 Drugs 5 received 200, 400 and 800 mg/kg oral doses of the
methanol extract of Laggera aurita (MELA)
Acetic acid (Ranbaxy Laboratories Ltd, Punjab); respectively. Thirty minutes after pretreatment, mice in
Ketoprofen (Lek; Slovenia); Piroxicam (Pfizer all groups were treated with acetic acid (0.6% v/v, 10
laboratories, Pakistan); Naloxone (Pfizer laboratories, ml/kg body weight i.p.). Mice were then placed in
Pakistan); Morphine sulphate BP (Martindale individual cages and the number of abdominal writhes
Pharmaceuticals, Ramford, Essex). All drugs or reagents was counted for each mouse for a period of 10 minutes
were freshly prepared to the desired concentration with after 5 minutes latency period.
distilled water or normal saline just before use.
Hot plate test in mice
2.5 Phytochemical Screening of Laggera aurita
The method previously described by Eddy and
Phytochemical screening was carried out on the Leimbach (1953) was employed for the study. Mice
methanol extract of the plant L. aurita using standard were grouped into five groups of six mice each. Group 1
protocol as described by Trease and Evans (1983). received 10 ml/kg normal saline orally. Groups 2, 3 and
4 received 200, 400 and 800 mg/kg of MELA orally
2.6 Acute Toxicity Studies in Mice and Rats (LD50) respectively, while group 5 received 5 mg/kg morphine
solution orally. Thirty minutes after treatment, each
LD50 determination was conducted using Organization mouse was placed on a hot plate (Gallenkamp
for Economic Co-operation and Development (OECD thermostat) which was set and maintained at 55 ± 1oC.
420) guidelines in rats and mice. In this method, two

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 Shehu et al, Afr. J. Pharmacol. Ther. 2016. 5(2): 65-73

The pain response latency was determined using a stop vascular congestion with lymphocyte hyperplasia in
watch. rats (Plate II).

Effect of naloxone pretreatment on analgesic effect Kidneys: The normal saline group showed normal renal
of methanol extract of Laggera aurita in the tissue, normal glomeruli, tubular epithelium, interstitial
thermally induced pain model tissue and vessels. At 2000 mg/kg of MELA, there was
slight glomerular necrosis in mice, slight glomerular
Four groups of 10 mice each were used. Initially all the necrosis and tubular distortion in rats. At 5000 mg/kg,
animals received naloxone at a dose of 2 mg/ kg i.p. there was slight glomerular necrosis with tubular
After 15 minutes, group 1 and 2 received 400 and 800 damage in mice (Plate III) and rats (Plate IV).
mg/ kg of MELA orally. Group 3 received normal saline
10 ml/kg orally and group 4 received morphine 5 mg/ Lungs: The saline groups showed normal alveoli in both
kg orally. The index of pain response latency was mice and rats. At 2000 mg/kg, alveoli were normal in
assessed at 60, 120 and 180 minutes using hot plate both mice and rats. At 5000 mg/kg, there was slight
method (Younos et al. 1990). alveoli congestion in mice (Plate V), while vacuotation
with lymphocyte hyperplasia and congestion were
2.9 Anti-inflammatory activity observed in rats (Plate VI).

Formalin induced inflammation Spleen: The saline group and at 2000 mg/kg extract,
red and white pulp in both mice and rats were normal.
Thirty rats were divided into five groups each of six At 5000 mg/kg there was lymphocyte hyperplasia
rats. Group 1 was treated with 10 ml/kg normal saline (Plate VII).
orally; group 2 was treated with 10 mg/kg ketoprofen
orally; while group 3, 4 and 5 received 200, 400 and 800 Stomach: The normal saline group showed normal
mg/kg of MELA respectively orally. Inflammation was mucosa lining in both mice and rats. At 2000 mg/kg,
induced in rat hind paw by injecting formalin 0.1 ml both mice and rats showed normal mucosa lining. At
(1% w/v in 0.9% normal saline) into the subplanter 5000 mg/kg, moderate erosion of stomach epithelium
surface of the left hind paw 30 mins after treatment. in mice and slight mucosa damage in rats (Plate VIII)
Paw thickness (mm) was measured using a digital were observed.
vernier caliper (MR 2002) at 0, 1, 2, 3, 4 and 5 hours
after formalin injection (Winter et al. 1963). Heart: Cardiac muscles in both mice and rats were
normal in the normal saline group, at 2000 mg/kg and
2.10 Data Analysis 5000 mg/kg of MELA.

The results were expressed as Mean ± SEM and 3.2 Analgesic activity
analyzed using one way ANOVA for acetic acid test and
repeated measures ANOVA for hot plate test, interaction Effect of methanol extract of L. aurita on acetic acid
of extract with naloxone, and formalin induced induced writhes test in mice
inflammation, followed by Bonferoni post hoc test.
Results were considered significant at P ≤0.05. MELA significantly (p<0.05) decreased the number of
acetic acid-induced writhes in mice in a dose dependent
3. Results and Discussion manner. The effect of MELA at all doses tested showed
significant (p<0.01) decrease in number of acetic acid-
3.1 Phytochemical screening induced writhes than piroxicam at a dose of 20 mg/kg
(Figure 1).
The phytochemical constituents found in the methanol
extract of L. aurita include the flavonoids, alkaloids, Effect of the methanol extract of L. aurita on
glycosides, saponins, cardiac glycosides, phenols, thermally induced pain
tannins, steroids and carbohydrates.
MELA significantly (p<0.05) increased the mean
3.2 Acute Toxicity Study reaction time in a dose dependent manner. At 800
mg/kg, MELA showed significant (p<0.01) increase in
The oral acute toxicity (LD50) was found to be greater mean reaction time than morphine (5 mg/kg) at three
than 5000 mg/kg body weight in both mice and rats. and four hours treatment time (Table 1).

Histological studies Effect of methanol extract of L. aurita interacted

with naloxone and assessed by hot plate test
Liver: The groups that received normal saline showed
normal hepatocytes and intact portal tract in both mice MELA co-administered with naloxone significantly
and rats. At 2000 mg/kg of the methanol extract of (p<0.01) decreased the reaction time in a dose
L.aurita (MELA), there was slight vascular congestion in dependent manner. At 800 mg/kg, MELA showed
mice and there was intense vascular damage with significant (p<0.001) decrease in reaction time than
kupffer cell hyperplasia and slight hepatic necrosis in morphine 5 mg/kg at the three hours treatment time
rats. At 5000 mg/kg of MELA, there was intense (Table 2).
vascular congestion with kupffer cell hyperplasia and
lymphocyte hyperplasia in mice (Plate I) and there was

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 Shehu et al, Afr. J. Pharmacol. Ther. 2016. 5(2): 65-73

Plate I: Photomicrograph of a liver section of mouse treated Plate II: Photomicrograph of a liver section of rat treated
with 5000 mg/kg of methanol extract of L. aurita and with 5000 mg/kg of methanol extract of L. aurita and
observed for fourteen days showing A = intense vascular observed for fourteen days showing A = vascular congestion,
congestion, B = Kupffer cell hyperplasia, C = lymphocyte B = lymphocyte hyperplasia. H & E stain (× 400)
hyperplasia. H & E stain (× 400)

Plate III: Photomicrograph of a kidney section of mouse Plate IV: Photomicrograph of a kidney section of rat treated
treated with 5000 mg/kg of methanol extract of L. aurita and with 5000 mg/kg of methanol extract of L. aurita and
observed for fourteen days showing A = slight glomerular observed for fourteen days showing A = slight glomerulus
necrosis, B = tubular damage. H & E stain (× 400) necrosis, B = tubular damage. H & E stain (× 400)

3.3 Anti-inflammatory activity edema and blue color denotes significant statistical
decrease in mean paw edema size. Peak of inflammation
Effect of the methanol extract of L. aurita on was reached in the third hour in normal saline and
formalin induced inflammation in rats extract dose 200 mg/kg, and in the second hour for
Ketoprofen 10 mg/kg, MELA 400 and 800 mg/kg. The
Formalin (0.6% v/v) produced local edema in rats paw extract significantly (p<0.001) decreased formalin-
which was significant as compared to time zero in all induced paw edema dose dependently at the fourth and
the groups (p<0.001). The red color denote peak of fifth hours post treatment time (Table 3).

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 Shehu et al, Afr. J. Pharmacol. Ther. 2016. 5(2): 65-73

Plate V: Photomicrograph of a lung section of mouse treated Plate VI: Photomicrograph of a lung section of rat treated
with 5000 mg/kg of methanol extract of L. aurita and with 5000 mg/kg of methanol extract of L. aurita and
observed for fourteen days showing A = slight alveoli observed for 14 days showing A = vacuotation, B =
congestion. H & E stain (× 400) lymphocyte hyperplasia, C = congestion. H & E stain (× 400)

Plate VII: Photomicrograph of a spleen section of mouse Plate VIII: Photomicrograph of a stomach section of rat
treated with 5000 mg/kg of methanol extract of L. aurita and treated with 5000 mg/kg of methanol extract of L. aurita and
observed for fourteen days showing A = lymphocyte observed for fourteen days showing A = slight mucosa
hyperplasia. H & E stain (× 400) damage. H & E stain (× 400)

4. Discussion phosphodiesterases (Middleton, 1998). Flavonoids

were also reported to be effective against inflammation
The biological or pharmacological actions of plant (Narayana et al. 2001) by inhibiting enzymes involved
extracts are known to be due to the presence of specific in prostaglandin synthesis (Kamil, 1993; Kim et al.
phytochemical constituents. Analgesic and anti- 2000).
inflammatory effects have been observed and reported
with flavonoids (Amin et al. 2012) and tannins Thus, the antinociceptive and anti-inflammatory effect
(Ahmadiani et al. 2000). There are also reports on exhibited by the plant L. aurita may be due to the
alkaloidal analgesic effects (Reanmongkol et al. 2005), presence of saponins, flavonoids, alkaloids,
and with saponins (Choi et al. 2005, Arrau et al. 2010). carbohydrates, tannins or combination of all found
Certain flavonoids possess potent inhibitory activity present in the plant.
against many enzymes such as protein kinase C, protein
tyrosine kinases, phospholipase A2 and

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 Shehu et al, Afr. J. Pharmacol. Ther. 2016. 5(2): 65-73

Data was analyzed using one way ANOVA followed by Bonferoni Post Hoc test and presented as mean
± SEM, * = p<0.05, ** =p<0.01, *** =P<0.001, **** =P< 0.0001 significant statistical difference as
compared with Control group, n=6, MELA= methanol extract of L. aurita, PIR = piroxicam 20 mg/kg
Figure 1: Effect of methanol extract of L. aurita on acetic acid induced writhing in mice

Table 1: Effect of the methanol extract of L. aurita on thermally induced pain in mice

Groups Mean reaction time ± SEM (secs)

(mg/kg) 0 min 30 mins 60 mins 90 mins 120 mins

Control 2.29±0.10 2.18±0.22 2.19±0.23 2.43±0.28 2.49±0.19

LA(200) 1.67±0.14 3.25±0.39 3.32±0.14** 3.01±0.24* 3.02±0.29*

LA(400) 1.81±0.25 2.82±0.14** 3.02±0.16 3.28±0.31 3.29±0.18

LA(800) 2.08±0.20 2.99±0.19 2.69±0.19 3.87±0.27** 3.81±0.24**
Mor (5) 2.15±0.19 3.37±0.20* 3.07±0.41 3.73±0.16* 3.72±0.27*
Data was analyzed using repeated measures ANOVA followed by Bonferroni post hoc test, * = P< 0.05, ** = P< 0.01 significant statistical
increase in mean reaction time compared with time zero; n=6. Values are Mean ± SEM. LA= methanol extract of Laggera aurita, Mor =
morphine 5 mg/kg

Table 2: Effect of methanol extract of L. aurita interacted with naloxone and assessed by hot plate test

Treatment Mean reaction time ± SEM (secs)

(mg/kg)` 0 mins 60 mins 120 mins 180 mins

Control 2.09±0.17 2.00±0.14 1.97±0.09 2.04±0.14

LA (400) 1.75±0.13 1.90±0.13 1.81±0.09 1.69±0.07
LA (800) 2.16±0.19 1.99±0.21* 2.03±0.19* 1.88±0.18**
Mor (5) 2.21±0.12 2.13±0.16 2.05±0.11 1.94±0.10*
Data was analyzed using repeated measures ANOVA followed by Bonferroni post hoc test, * = p<0.01, ** = p<0.001 significant statistical
difference compared with time zero; n=10. Values are Mean ± SEM. LA= methanol extract of L. aurita, Mor= morphine 5 mg/kg

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 Shehu et al, Afr. J. Pharmacol. Ther. 2016. 5(2): 65-73

Table 3: Effect of methanol extract of L. aurita on formalin induced inflammation in rats

Treatment Mean Paw Diameter (millimetre)

(mg/kg) 0 hrs 1 hrs 2 hrs 3 hrs 4 hrs 5 hrs

Control 2.24±0.06 2.78±0.10## 3.18±0.05## 3.4±0.05#### 3.03±0.06#### 2.7±0.06#

LA (200) 2.08±0.05 2.76±0.06## 2.73±0.04## 2.88±0.04## 2.57±0.03**## 2.46±0.01*##
LA (400) 2.19±0.07 2.86±0.02## 2.87±0.02## 2.86±0.06### 2.65±0.04** 2.34±0.05**
LA (800) 2.21±0.09 2.86±0.03# 2.82±0.04# 2.78±0.03## 2.46±0.06** 2.33±0.04**
Ket (10) 2.21±0.05 2.55±0.06## 2.69±0.09## 2.32±0.04## 2.3±0.02** 2.19±0.01**
Data was analyzed using repeated measures ANOVA followed by Bonferoni Post Hoc test, *= p<0.05, **= p<0.0001, significant statistical
decrease in mean paw edema size (blue) as compared to the peak of edema (red), # = p<0.05, ## = p<0.01, #### = p<0.000 significant
statistical increase in mean paw diameter as compared to time zero; n= 6. Values are Mean ± SEM. LA= methanol extract of L. aurita,
Ket= ketoprofen 10 mg/kg, hrs= hours

The oral LD50 was above 5000 mg/kg for MELA, while the second phase is mainly attributed to the
showing relative safety. LD50 is a useful index in release of prostaglandins (Vinegar et al. 1969; Chan et
assessing the safety margin of a substance. The OECD al. 1995). MELA inhibited both phases of inflammation
(Walum, 1998) classification of acute systemic toxicity which may be due to its ability to inhibit the release of
based oral LD50 of >500 ≤ 2000 mg/kg as non-toxic or the mediators of inflammation.
harmful. Based on this, the oral LD50 greater than 5000
mg/kg established in both mice and rats indicated Some analgesic drugs act by inhibiting prostanoids
relative oral safety. Other toxicity scales (Hodge and biosynthesis. They act as analgesic and anti-
Sterner, 1943) reported that compound with an oral inflammatory agents by inhibiting COX-2 dependent
LD50 of between 500 – 2000 mg/kg should be prostanoids in the cells at inflammatory sites (Mishra et
considered practically non-toxic. al. 2011) and in the spinal cord (Ramer et al. 1998).
They also inhibit the action of phospholipase A2, which
Acetic acid induced writhing test is usually employed to releases arachidonic acid from the cell membrane
screen for peripheral analgesic activity (Bentley et al. (Burke et al. 2006). These classes of drugs are usually
1981, Gene et al. 1998). Intraperitoneal administration associated with side effects like stomach problems such
of acetic acid to mice leads to the release of as bleeding, ulcer, stomach upset (Traversa et al. 1995),
prostaglandins like PGE2 and PGF2α which have been kidney problems, heart problems (Gislason et al. 2009),
implicated as mediators of pains and inflammations high blood pressure and fluid retention (Knight et al.
(Santosh et al. 2008). Thus, activity shown by MELA 2010). Thus, the histological changes observed in the
suggests that analgesic effects may be due to their organs from animals exposed to 2000 and 5000 mg/kg
action on visceral receptors sensitive to acetic acid, or may be resultant from inhibition of prostanoid
inhibition of the production of algogenic substances, or biosynthesis.
the inhibition at the central level of painful messages
transmission or combination of all the three mentioned 5. Conclusion
above.
The methanol extract of L. aurita has been shown to
The thermally induced pain model is utilized as a possess significant analgesic and anti-inflammatory
standard method for the evaluation of centrally activities which may be responsible for it use in painful
mediated analgesia. Narcotic agents like morphine, and inflammatory conditions. Based on this, studies
pentazocine and codeine mediate their analgesic effect should be carried out to isolate, characterize and
through this mechanism (Leonard et al. 2006). The elucidate the structure of the bioactive constituents
analgesic activity observed in MELA may be due to its responsible for the observed pharmacological effects.
activity via central mechanism similar to the narcotic
analgesic agents (Vogel and Vogel, 1997). This centrally The results of histological studies showed effects on
mediated effect which is through interaction with kidney and liver at dose of 2000 mg/kg and on kidney,
opioid receptors (Leonard et al. 2006) was blocked by liver, spleen, lungs and stomach at dose of 5000 mg/kg,
prior administration of naloxone. Naloxone is a specific thus detailed toxicological screening on long term
antagonist of opioid receptors (Stein, 1995, Almeida et repeated dosing should be conducted.
al. 2011), and thus the inhibition of the analgesic action
in the presence of this agent suggests possible central
and opioid receptor involvement in the analgesic effect
of the plant extract. Conflict of Interest Declaration

Formalin induced inflammatory response is usually bi- The authors declare no conflict of interest.
phasic in nature. The first phase is mainly due to the
release of histamine, serotonin and kinins (1-3 hours)

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 Shehu et al, Afr. J. Pharmacol. Ther. 2016. 5(2): 65-73

Acknowledgement Gene RM, Segura L, Adzet T, Marin E, and Ingelsias J (1998).

Heterotheca inuloides: Inflammatory and Analgesic Effects. J.
The authors acknowledge the effort of Dr Jamilu Yau for Ethnopharmacol. 60: 157-162.
providing us with pure naloxone powder, Ibrahim
Adamu for providing us with laboratory animals and all Gislason GH, Rasmussen JN, Abildstran SZ, Schramm, TK,
technical staff of Pharmacology Department, Ahmadu Hansen ML, Fosbol EL, Sorensen R, Poulsen HE, Kober L,
Bello University, Zaria, Nigeria. Madsen M, and Torp-Perdesen C (2009). Increased Mortality
and Cardiovascular Morbidity Associated with use of Non-
Steroidal Anti-inflammatory Drugs in Chronic Heart Failure.
Arch. Intern. Med. 150: JC6-12.
References
Hodge HC and Sterner JH (1943). Determination of substances
Abdulla MA, Lutfi MF, Baket, AO, and Mohammed AH (2013). acute toxicity by LD50 Am. Ind. Hyg. Assoc. 10: 93-98.
Evaluation of the Anti-inflammatory Effects of Blumea aurita.
Global J. Med. Res. (B), 13: 1-8. Kamil M (1993). Glimpses in plant research. Vol XI, Today and
Tomorrow’s Printers and Publishers, New York, U.S.A Pp 377-
Ahmadiani A, Hosseiny J, Semnanian S, Javan M, Saeedi F, 389.
Kamalinejad M and Serami S (2000). Antinociceptive and Anti-
inflammatory effects of Elaeagnus angustifolia fruits extract. J. Kim HP, Son KH, Chang HW, and Kang SS (2000). Effects of
Ethnopharmacol, 72: 287-292. naturally occurring Flavonoids on Inflammatory Response
and their action Mechanism. Nat. Prod. Sci. 6: 170-178.
Almeida RN, Navarro DS, and Barbosa-Filho JM (2001). Plants
with central analgesic activity. Phytomedicine. 8: 310-322. Knight MK, Mangoni AA, and Miners JO (2010). Defining the
COX inhibitor Selectivity of NSAIDs: Implication for
Amin E, Abouzid S, and Seida A (2012). Phytochemical and Understanding Toxicity, Expert Rev. Clin. Pharmacol. 3:769-
Biological Screening on Isoflavonoids from Dalbergia 776.
paniculata Cultivated in Egypt. Pharmacologia, 3: 84-90.
Koster RA, Anderson M, and De-beer EJ (1959). Acetic acid for
Arrau S, Delporte C, Cartegena C, Rodriguez-Diaz M, Gongalez Analgesic Screening. Fed. Proc. 18: 412.
P, Silva X, Cassels BK and Miranda HF (2010). Antinociceptive
activity of Quillaja saponaria Mol. Saponin extract, quillaic acid Leonard JP, Daniel SB, and Yongli Xi MS (2006). Increasing
and derivatives in mice. J. Ethnopharmacol, 133: 164-167. Death from Opioid Analgesics in the United States.
Pharmacoepidemiol. Drug Saf. 15: 618-627.
Arthur SJ and John B (1978). A colour Atlas of
Histopathological Staining Techniques. Wolf MED.Pub. Ltd. Middleton E (1998). Effect of flavonoids on immune and
London, pp 14-20. inflammatory cell function. Adv. Exp. Med. Biol. 439: 175-186.

Bentley GA, Newton SH, and Starr J (1981). Evidence for an Mishra D, Ghosh G, Kumar PS, and Panda, PK (2011). An
action of morphine and enkephalins on sensory nerve endings Experimental Study of Analgesic Activity of Selective COX-2
in the mouse peritoneum. Br. J. Pharmacol. 73: 325-332. Inhibitor with Conventional NSAIDs. Asian J. Pharm. Clin. Res.
4: 78-81.
Burke A, Smyth E, and Fitz GA (2006). Analgesic-Antipyretic
Agents; Pharmacotherapy of Gout. In: Brunton, L.L., Lazo, J.S OECD (2002), Test No. 420: Acute Oral Toxicity-Fixed Dose
and Parker, K.L. (Eds). Goodmann and Gilman’s, The Procedure, OECD guidelines for the Testing of Chemicals,
Pharmacological Basis of Therapeutics, 11th Ed. Mc Graw Co. Section 4, OECD Publishing, Paris.
Inc. New York, Pp 671-715.
Olurishe TO and Mati FG (2014). Anti-hyperalgesic potentials
Burkill HM (1985). The Useful Plants of West Tropical Africa of Laggera aurita in Swiss Albino Mice. Pak. J. Pharm. Sci. 27:
Vol 1, 2nd Ed. Royal Botanical Gardens, Kew England, 452-453. 169-172.

Chan Y, Tsai HY, and Wu TS (1995). Anti-inflammatory and Narayana KR, Sripal RM Krishna DR, and Chaluvadi MR
Analgesic Activity of Extracts from the Root of Angelica (2001). Bioflavonoids Classification, Pharmacological,
pubescens. Planta Med. 61: 2-8. Biochemical effects and Therapeutic potential, Indian J.
Pharmacol. 33: 2-16.
Choi J, Jung H, Lee K, and Park H (2005). Antinociceptive and
Anti-inflammatory effects of Saponins and Sapogenan Ramer MS, Murphy PG, Richardson PM, and Bisby MA (1998).
obtained from the Stem of Akebia quinata. J. Med. Food. 8: 78- Spinal nerve lesion-induced mechanoallodynia and adrenergic
85. sprouting in sensory ganglia are attenuated in interleukin-6
knockout mice. Pain. 78: 115-121.
Eddy NB and Leimbach D (1953). Synthetic Analgesics II
Dithienylbutenyl and Dithienylbutylamines. J. Pharmacol. Exp. Reanmongkol W, Subhadhirasakul S, Thienmontree S,
Ther. 107: 385-393. Thanyapanit K, Kalnaowakul J, and Sengsui S (2005).
Antinociceptive activity of alkaloid extracts from Kopsia
Gedif T and Hahn HJ (2003). The Uses of Medicinal Plants in macrophylla leaves in mice. Songklanakarin J. Sci. Technol. 27:
self-care in Rural Central Ethiopia. J. Ethnopharmacol. 87: 509-516.
155-161.
Santosh SB, Sohan SC, Anupamaa S, Devanand S, and Manohar
JP (2008). Anti-inflammatory activity of an isolated Flavonoid

A KeSoBAP Publication ©2016. All rights reserved. ISSN 2303-9841

72
 Shehu et al, Afr. J. Pharmacol. Ther. 2016. 5(2): 65-73

fraction from Celosia argentea Linn. J. Med. Plants Res. 2: 052- Vikrant A (2011). A Review on Anti Tubercular Plants. Int. J.
054. Pharm. Tech. Res. 3: 872-880.

Shahwar D, Ahmad N, Khan MA, Ullah S, and Ahmad N (2012). Vinegar R, Schreiber W, and Hugo R (1969). Biphasic
Chemical Composition and Biological Activities of the essential Development of Carrageenan Edema in Rats. J. Pharmacol. Exp.
oil of Laggera aurita Linn (DC.) grown in Pakistan. Turk. J. Ther. 166: 96-103.
Biochem. 37: 329-335.
Vogel HG and Vogel W H (1997). Drug Discovery and
Stein C (1995). The control of pain in peripheral tissues by Evaluation: Pharmacological assays, Springer-Verlag, Berlin
opioids. New Engl. J. Med. 332: 1685-1690. Heidelberg. Pp 204 – 212.

Traversa G, Walker AM, Ippolito, FM, Caffari B, Capurso L, Dezi Walum E (1998). Acute Oral Toxicity. Environ. Health Persp.
A, Koch M, Maggini M, Alegiani SS and Raschetti R (1995). 106: 497-503.
Gastroduodenal toxicity of different Non-steroidal anti-
inflammatory drugs. Epidemiol. 6: 49-54. Winter CA, Risley EA, and Nuss GW (1963). Carrageenan
induced Edema in the hind paw of the rats as an assay for anti-
Trease GE and Evans WC (1983). Pharmacognosy 12th Ed. inflammatory drugs. Proc. Soc. Exp. Biol. Med. 111: 544-547.
Publ Bailere Tindall Macmillan, London UK.
Younos C, Rolland A, Fleurentin J, Lanhers MC, Misslin R, and
Usman H, Yaro AH and Garba MM (2008). Analgesic and Anti- Mortier F (1990). Analgesic and Behavioural effect of
inflammatory Screening of Newbouldia laevis flower in Morinda citrifolia, Planta Med. 56: 430-434.
Rodents. Trends Med. Res. 3: 10-15.

A KeSoBAP Publication ©2016. All rights reserved. ISSN 2303-9841

73