Professional Documents
Culture Documents
Analgesic Antipyretic And........
Analgesic Antipyretic And........
July, 2017
ii
iii
DEDICATION
education.
iv
ACKNOWLEDGEMENT
N.M Njagi and Dr Mathew Piero Ngugi for their inspiration, mentorship and
tireless effort in guiding me to complete my research study. May God bless them
abundantly.
her staff for the close companionship and cooperation during the entire period of
mentioned, Mrs Lel, James Adino, Daniel Gitonga and James Ngunjiri for their
Mwonjoria, Jane Maoga, Wylcliffe Arika, Alex Cheruiyot, Herbet Cheruiyot and
I also owe a big thanks to my parents, brothers, sister and friends for the
encouragement, motivation and support throughout the entire period. Above all I
thank the almighty GOD for giving me good health and strength to complete this
project.
v
TABLE OF CONTENT
DECLARATION ............................................................................................. ii
DEDICATION ................................................................................................ iii
ACKNOWLEDGEMENT .............................................................................. iv
TABLE OF CONTENT ................................................................................... v
LIST OF FIGURES ...................................................................................... viii
LIST OF TABLES .......................................................................................... ix
LIST OF APPENDICES .................................................................................. x
ABBREVIATIONS AND ACRONYMS ....................................................... xi
ABSTRACT ................................................................................................... xii
CHAPTER ONE .............................................................................................. 1
INTRODUCTION ........................................................................................... 1
1.1 Background of the Study ........................................................................... 1
1.2 Problem Statement and Justification .......................................................... 7
1.3 Hypotheses ................................................................................................. 8
1.4 General Objective ...................................................................................... 9
1.4.1 Specific Objectives ................................................................................9
CHAPTER TWO ........................................................................................... 10
LITERATURE REVIEW .............................................................................. 10
2.1 Biochemical and Physiological Basis of Pain, Fever and
inflammation ............................................................................................ 10
2.2 Screening Models for Pain, Pyrexia and Inflammation ........................... 15
2.2.1 Screening Models for Pain ..................................................................15
2.2.1.1 Test Based on Thermal Stimuli..................................................15
2.2.1.1.1 Tail flick test using radiant heat ................................................15
2.2.1.1.2 Hot plate test ..............................................................................16
2.2.1.2 Test Based on Mechanical Stimuli ...............................................16
2.2.1.3 Test Based on Electrical Stimuli ..................................................16
2.2.1.3.1 Electrical stimulation of the tooth-pulp .....................................16
vi
LIST OF FIGURES
Figure 2. 1: Clutia abyssinica…………………………………………………..32
Figure 4. 1: Percentage change in rectal temperature of DCM root
extract of Clutia abyssinica on turpentine-induced
pyrexia in Wistar albino rats..............................................................49
LIST OF TABLES
LIST OF APPENDICES
ABSTRACT
Various diseases and injuries are always presented with pain, fever and
inflammation. These are considered as symptoms associated with various
pathological processes in an animal body. Drugs that are used to alleviate pain,
fever and inflammation such as non-steroidal anti-inflammatory drugs exhibit
adverse effects for example cardiac abnormalities, peptic ulcers, liver toxicity and
kidney failure. Therefore, there is need to come up with alternative remedies.
Herbal medicines are deemed to be safe, have good efficacy and have fewer side
effects. Clutia abyssinica is a shrub that is found in East, Central, and South
Africa and it has been used traditionally to cure several ailments including
malaria, chest pain, gonorrhea, fever, infertility, pain, inflammation, skin diseases
and cancer. Roots of this medicinal plant have been used traditionally to prepare
decoctions. The aim of the project was to evaluate the analgesic, antipyretic and
anti-inflammatory potential of dichloromethanolic root extract of Clutia
abyssinica in animal models. Plant sample material was collected from Kaptebee
village, Turbo sub-county in Uasin Gishu County Kenya, and the active
components extracted using dichloromethane. Pain, fever and inflammation were
induced Swiss albino mice and Wistar albino rats using acetic acid, turpentine and
carrageenan respectively. Swiss albino mice and Wistar albino rats were grouped
into normal control, negative control, positive control and 3 experimental groups.
Extracted root extracts were administered intraperitoneally to Swiss albino mice
and Wistar albino rats at predetermined doses (50, 100, and 150 mg/kg body
weight). The analgesic and anti-inflammatory activities of the plant root extract
were compared to diclofenac the (reference drug) while the antipyretic activity
was compared to aspirin. The dichloromethanolic root extract of C. abyssinica
demonstrated significant analgesic, antipyretic and anti-inflammatory activities.
Number of abdominal writhing was reduced between 33.95-49.51% by
dichloromethanolic root extract while the diclofenac (reference drug) reduced
abdominal writhing by 46.51%. Reduction in number of abdominal writhings by
the extract indicates the plant posse‟s analgesic properties. The elevated
temperature was reduced between 0.68-3.34% by the dichloromethanolic root
extract while Aspirin the (reference drug) reduced elevated temperature between
3.32-4.96%. Edema was reduced between 0.88-5.34% by the plant extract while
diclofenac reduced edema between 2.21-5.35% respectively. Rectal temperature
and the size of the edema was reduced more in the third and fourth hours
signifying better blockage of mediators responsible for fever and inflammation.
Data was analyzed using one way analysis of variance followed by turkey‟s test.
Qualitative phytochemical analysis revealed the presence of alkaloids, flavonoids,
terpenoids, steroids, saponins and cardiac glycosides. The extract from Clutia
abyssinica may be used as an alternative bioresource in development of analgesic,
antipyretic and anti-inflammatory agent. The study therefore, confirms the
folklore use of the medicinal plant by Kalenjin community of Kenya to manage
pain, fever and inflammation.
1
CHAPTER ONE
INTRODUCTION
intensity, space, emotion, cognition and motivation (Maze et al., 2000). Pain
histamine, serotonin and prostaglandins. Pain in its real sense lack a way to
define it, but in general term, occurs whenever the body tissues are damaged
(Arome et al., 2016). Sensation of pain is a sign that something in the body is
wrong. Pain plays an important role in drawing attention to tissue injury from
harmful stimuli and reflexes are elicited to protect the injured part of the body
mediated by nervous system and is used for diagnosing various diseases such as
diabetes, arthritis and cancer that are normally associated with chronic pain
temperature beyond the normal range of (36.5°C -37.5°C) (Axelrod and Diringer,
chills, sensation of cold and other subjective sensations (Guyton and Hall, 2000).
A number of different microorganisms and other substances can cause fever and
are collectively termed as pyrogens (Shalini and Donna, 2006). Pyrogens are
those that originate outside the body for example bacteria toxins on the other hand
endogenous pyrogens are derived from immune cells in the body responding to
breakdown of protein products in an organism body can cause the set point of the
health conditions the range for oral temperature is between 33.2 – 38.2°C, for the
armpit 35.5 – 37.0°C, the rectal 34.4 – 37.8°C, while for tympanic membrane
3
injury and this include; leukocyte infiltration, edema formation and granuloma
formation (Guyton and Hall, 2000; Mitchell and Cotran, 2000). For edema to be
increase vascular permeability and blood flow into the damaged sides (Lalenti et
considered as that initial response to harmful stimuli by the body while chronic
2003).
pain, and loss of function (Craig and Stitzel, 2003). A diverse number of
histamine, serotonin, kinins and platelet activating factors (Burke et al., 2006).
Pain, fever and inflammation are beneficial to the immune system. However, they
cause a lot of suffering and discomfort to the victims lowering the quality of life
(NSAIDs) are commonly used to manage inflammation, fever and pain (Barar,
2009). Opoid analgesics are choice drugs for severe or chronic malignant pain
analgesics that are normally used to treat inflammation, fever and pain manifest a
there is still need for development of more cost-effective and improved remedies
with lesser side effects. Recent studies by world health organisation (WHO)
inflammation and fever manifest a lot of side effects after long term use that
tendon, cartilage healing and delay in muscle regeneration in many studies has
manage pain, inflammation and fever only provide asymptomatic relief and the
greatest disadvantage lies in their toxicity to the liver, kidney and reappearance of
alternative medicine (CAM) for treatment of pain, fever and inflammation as well
only a small percentage are included in health care systems after clinical research
artemisinin were derived from plant products, the search for plant species with
al., 2014). Though there are some ethnobotanical studies conducted on this
medicinal plant, there has been no scientifically evaluated data about its potential.
anti-inflammatory drugs.
7
treatment and management of many serious indicators of ill health including pain,
fever and inflammation is still problematic and complex (Adedapo et al., 2009).
Pain, fever and inflammation cause suffering and discomfort among the victims
(Kariuki et al., 2012). Recent studies by WHO indicates that the NSAIDs used in
management of these conditions manifest a lot of side effects (Robotin, 2006; Beg
et al., 2011).
safe, have good efficacy, are culturally accepted and have lesser side effects than
Uasin Gishu County Kenya by the Kalenjin community to manage pain, fever,
extensive search on the literature reveals that no data has been documented about
the medicinal use of the plant against pain, fever and inflammation.
8
1.3 Hypotheses
mice models.
ii. The DCM root extracts of C. abyssinica do not have antipyretic effects in
rat models.
activities.
9
extracts of C. abyssinica.
10
CHAPTER TWO
LITERATURE REVIEW
The international association for the study of pain (IASP) defines pain as
tissue damage”. The emotional component differ from one person to the other
and in the same individual from time to time and it can be classified in several
2006). In the body, Sensory nerve endings are generally found in every part of the
body such as the blood vessels, internal organs, muscles, joints, and the skin.
released which then activate and sensitize nociceptors to other mediators of pain
stimuli is initiated by peripheral receptors (Guyton and Hall, 2006). In the brain
pain stimulus are processed and generated impulses are send down the spinal cord
following the appropriate nerves and instructs the body to respond, for instance
Peripheral nerves transmit pain stimulus to the spinal cord which then links to the
brain. Two types of nerve fibers are involved in this process; slow pain fibers and
Fast pain fibers. Transmission of fast pain is through the A delta fibers (Aδ fibers)
11
to the spinal cord. The activity of fast pain fibers is terminated at luminal in the
pathway and terminates its transmission in the brainstem (Arome et al., 2016).
transmits fast pain impulses to the brain in the cortex. Therefore localization of
pain in certain part of the body becomes relatively precise (Rang et al., 2006).
pain (Craig and Stitzel, 2003). It is a sensory modality that is essential for survival
system to initiate a response that would otherwise minimize injury to the tissues
Pain can also be classified into fast and slow pain. A second after application of
pain stimulus such as pricking of hand or touching a hot object fast pain is felt at
that moment. This kind of pain is shallow, felt within underneath of the skin but
not felt in most internal tissues of the body. Its transmission is through the A delta
fibers at a velocity of about 5-30 m/s and due to its high speed in conduction of
pain stimulus it allows the body to immediately withdraw from the stimuli
(Guyton and Hall, 2006). Slow pain on the other hand is throbbing and diffused.
It is felt immediately after pain stimulus is applied lasting for few minutes, weeks
or even months and if not properly processed by the body, may result in chronic
12
pain. Slow pain is felt in internal tissues of the body and its transmission is
through C-fibers at a speed of 0.5-2 m/s to the brain. Other types of pain include
the visceral, somatic and neuropathic pain (Guyton and Hall, 2006).
Fever means the body temperature is above normal and fever is a symptom, not a
infections. Fever turns on the body‟s immune system and helps fight infections
systemic response is generated which act on the rest of the body, causing heat-
creating effects to match the new temperature. The hypothalamus works like a
In essence, endogenous pyrogens are cytokines which are associated with innate
immune system. Phagocytic cells produce this molecules that causes increase in
(Walter, 2003). When the set-point is raised, the body develop‟s mechanism of
heat generation and retention. Vasoconstriction decreases loss of heat through the
skin and makes an individual to feel cold (Anochie and Philip, 2013). This differs
Febrile response involves innate immune system activation via Toll-like receptor
and tumor necrosis factor (TNF-α) that act on an area of the brain known as the
OVLT and eventually leading to the release of PGE2 via activation of COX-2
enzyme (Young and Sexana, 2014). In the hypothalamus, PGE2 binds to receptors
until a new elevated set-point in the hypothalamus is reached (Young and Sexana,
2014).
which blood-derived products such as plasma proteins, fluids and leukocytes are
breakdown, enzyme activation, regeneration and repair (Vane and Bolting, 1995).
There are two kinds of inflammation, acute and chronic inflammation. Acute
inflammation begins immediately after the injury of tissues and is usually marked
by cardinal signs such as redness, heat, pain and loss of function. On the other
(Lange et al., 2001; Proell et al., 2008). Once ligands are recognized, TLRs
activate ordinary signaling pathways and these results in the activation of NF-κB
(Ghosh et al., 1998). When the signal is transducted, NF-κB is released from IκB
receptors that alert the immune system to cell injury and provide ways in which
cytokines that promote inflammation that include TNF-α, IL- 1β, IL-6, and others.
as reactive oxygen species and nitrogen species. Both oxygen and glucose are
consumed when these noxious chemicals are released from the cytoplasmic
Prostaglandins act as short-lived localized hormones that are released when any
cell in the body is exposed to traumatic injury or any other form harmful stimuli.
Once present in the intracellular space prostaglandins can induce fever, pain and
15
regulation of blood vessel tone, clot formation and platelet aggregation (Rehman
Pain models have been classified into: Thermal, electrical, mechanical and
chemical stimuli according to the kind of stimuli applied. The neuronal basis of
this models is not clearly known, however they are used to predict analgesic
thermal radiation is applied to the tail/paw of the animal it triggers the animal to
withdraw its tail/paw from the thermal source (Smith et al., 1943). Tail
withdrawal from the heat source is referred to as “tail flick latency‟‟. Time taken
for the animal to withdraw its paw/tail from the heat source in this model is timed
and recorded.
16
surface consisting of hot metal or boiling liquid by a thermode. The time taken
during paw licking and jumping from the heat source is timed and recorded, this
is called reaction time. Both licking of the paw and jumping from heat source are
The hind paw and the tail are ideal sites for applying nociceptive mechanical
stimuli. In this model the tail or paw is jammed between two plane surfaces and
the pressure of increasing intensity is applied until the animal begins a response
behavior of withdrawing its tail or hind paw from the two planes. The vocal
reaction taken by the animal to withdraw tail or hind paw from two the plane
model. This produces behavioral characteristic reaction such as head flick, biting,
chewing, and licking of the tooth pulp due to induction of pain. Time taken for the
In this model electrical current of increasing intensity is applied to the tail of the
rat or mice. This will generate some observed reflex movement in the tail with
increasing electrical currents. Time taken for the animal to withdraw its tail from
electrical stimuli is timed and recorded. Morphine or drugs falling in the same
class as morphine are effective in this model (Parle and Yadav, 2013).
prostaglandins (Turner, 1965). When formalin is injected into the hind paw of the
animal it elicits a painful behavior such as licking of the paw, bitting and lifting.
The time taken by the animal to lick, lift and bit the paw is timed and recorded
In this model acetic acid or phenylquinone is used to induce pain in mice or rats
by injecting these irritants into the peritoneal cavity. The animal responds by
musculature contraction and stomach touching the floor. This model is sensitive
to peripherally acting analgesics. This agent irritates the serous membrane and
18
Turpentine induces fever slowly reaching its peak at 11th hour after its injection.
without generating adenosine triphosphate causing the release of calcium from its
mitochondrial stores and subsequently prevent the uptake of calcium. This leads
and this will in turn result in hypothalamus producing prostaglandins (Dubey and
Carrageenan has widely been used as a harmful agent for inducing inflammation
activities. When injected into experimental animal this phlogistic agent produces
known to induce inflammation in two phases and is dependent upon age and
dependent on age and weight of the experimental animal (Vinegar et al., 1969).
The first phase (0-2 hours) is due to release of inflammatory mediators such as
20
The lysosome, protease, prostaglandins and bradykinin are released majorly in the
second phase (D‟Amour et al., 1965). The second phase (2.5-5 hours) of edema is
prostaglandins play a major role in inflammatory reaction and can stimulate the
nociceptors and thus induce pain (Dacie, 1958). The initial phase is due to release
et al., 1969).
This technique is based on the ability of the drug to hinder inflammation produced
in the hind paw of the mice or rat after injection with formalin. Nociceptive effect
release of histamine, serotonin (5-HT), and kinin while the second phase is
in the dorsal region to induce granulomas lesion. This model is used to asses
therapeutic agents that are prescribed all over the world for treatment of
analgesic activities (Lascelles et al., 2007; Sparkes et al., 2010). These NSAIDs
for instant aspirin is known to acetylate the active site of COX enzyme called
Tyro 385 and Ser 530. Aspirin and arachidonic acid compete for the active
22
prostaglandins synthesis (Rao and Knaus, 2008). Generally NSAIDs exhibit three
ii. Competitive, reversible inhibitors, time dependent that bind COX active
site in the early phase to form a reversible enzyme inhibitor complex for
During the early 18th century, salicin a compound responsible for the synthesis of
acetylsalicylclic acid was discovered and isolated. In the 1990s there was the
therapies to manage pain, pyrexia and inflammation (Marnett, 2009). NSAIDs for
term fever, pain and inflammation (Inotai et al., 2010). Analgesic drugs such as
Acetylsalicylic acid is known for its ability to irreversibly disable COX enzymes
also act by suppressing the production of pyrogenic cytokines such as TNF-α and
1b, IL-1a, TNF-α and IL-6 (Ghosh et al., 1998). New generation of NSAIDs
reduced side effects (Boursinos et al., 2009). Ability of NSAIDS to disrupt the
inflammation and pain has spread (Hawkey and Rampton, 1985). Conventional
NSAIDs that work as analgesic, antipyretic and anti-inflammatory agents are non-
specific cyclooxygenase enzyme blockers on the other hand selective NSAIDs are
specific in their inhibition by inhibiting COX-2 only (Vane et al., 1998). Non-
COX-2 (Vane et al., 1998). COX 1 enzyme plays an important role in renal and
gastrointestinal blood flow as well platelet aggregation while COX 2, on the hand
plays a key role in pain and inflammation process (Rao and Knaus, 2008).
ibuprofen, and naproxen are usually used for management of pyrexia, pain and
inflammation (Paya and Katzung, 1995). NSAIDs represent a class of drugs that
are used to prevent and treat postoperative pain (Luna et al., 2007). However,
formed. Its negative effects are linked to the inhibition of prostaglandin which has
an effect on oesteoblasts (Boursinos et al., 2009). It has been shown that long
in ulnar osteotomy (Waters et al., 2000). Recent studies by WHO indicates that
these conventional drugs used in the management of pain, inflammation and fever
25
manifest a lot of side effects after long term use for example gastric irritation,
Traditional medicinal herbs for over centuries have served as potential source for
alternative medicine and the knowledge of herbal medicine has been passed on
Greek, Syrian, African and Roman medicinal practices documented the use herbal
medicine for curing different diseases (Komboj, 2000). Many communities all
over the world have utilized Phytopharmaceuticals for centuries in curing an array
of ailments (Semwal et al., 2010). Herbal medicines from medicinal plants have
(Shinwari, 2010).
(Vasundra and divya, 2013). Nature is endowed with complete store of remedies
to cure all diseases affecting the human being (Kokate et al., 2002). Nature is
also endowed with natural remedies inform of herbs, animal products and algae
capable of curing diseases without any side effects (Trease and Evans 1983).
26
to relieve inflammation, pain and fever should be seen as a step towards discovery
herbal medicines (Gupta et al., 2006). Secondary active constituents from plant
sources are directly used as therapeutic agent and can serve as lead molecule in
2002 by the National Center for complementary and alternative medicine (CAM)
surveyed a total of 31,044 adults (Barnes et al., 2004). Found out that during the
2.5.1 Pain
Throughout history man has used different forms of therapy to relief pain.
Morphine for example was isolated from a medicinal plant Papaver somniferium
(De souza, 2011). The search of herbal plants with analgesic activities, used as
pain relievers should be viewed as a successful search for new pain relieving
artemisinin, atrophine and chloroquine were derived from the plant products
White Willow Bark (Salix alba) has been used traditionally in management of
Salicin extract when taken at a dose level of 120-240 mg on daily basis can
associated with pain such as, low back pain, osteoarthritis, rheumatoid arthritis,
gastrointestinal upset gout, myalgia, chronic low back pain and lumbago.
albino mice models. The extract showed dose dependent response with 100 mg/kg
body weight having the highest inhibition percentage compared to 50 mg/kg body
when pain was induced using acetic acid, hot plate and tail flick test in laboratory
animals.
2.5.2 Pyrexia
Though medicinal plants from time immemorial have been used as a source of
resulted in neglect of their use. But due to its availability, low cost and fewer side
effects herbal medicine is gaining its popularity (Sharma et al., 2010). Treatment
of fever dates back to 400 B.C years ago when Greek Hippocrates prescribed an
extract from the willow bark and leaves (Rao and Knaus, 2008). Tumeric
(Curcuma longa) is an ancient spice that has been used traditionally as condiment,
According to Vadivelu et al. (2011), white Willow Bark (Salix alba) contains
has been atributed to flavonoids, tannins and salicylates present in the extract
capitata, the extract exhibited antipyretic activity in sprague dawley rats. Dose
level of 100 mg/kg and 200 mg/kg body weight extract reduced body temperature
2.5.3 Anti-inflammatory
For centuries, mankind has used herbal medicine for relieving inflammation and
pain (Sen et al., 2010). Boswellia serrata is among many herbal plants used in
(Curcuma longa) is an ancient spice and condiment that has been used as herbal
Gupta, 1972).
Camellia sinensis commonly known as tea plant is an important herbal plant. The
leaves and buds produce tea, the most consumed beverage in the world. The anti-
been reported (Curtis et al., 2004). Likewise, study carried out by Mwangi et al.
Clutia abyssinica (Figure 2.1) belongs to the genus Clutia and family of
Euphorbiaceae. It is dioecious, erect, lax shrub that grows up to 4–5 m tall, with
brittle branches and glabrous to evenly hairy. Leaves alternate, simple and entire.
Stem is green tinged red-brown; leaves are green but turn red when old.
usually harvested from the wild for its local medicinal use (Matu, 2008).
Clutia abyssinica (Figure 2.1) is found in Central Africa to Eritrea, Ethiopia and
through eastern Africa south to Zambia, Angola, Mozambique and South Africa.
abyssinica is commonly found in dry forest, forest remnants, secondary forest and
altitude level of between 700–3700 m above the sea level (Matu, 2008).
32
extensively used as herbal medicine in South Nandi District Kenya and it has
been used traditionally to treat several ailments that include; malaria, colds, fever,
skin diseases, chest problems, cancer and infertility in humans (Jeruto et al.,
2011). The root and leaf extracts are drunk or rubbed on the head to treat
headache. Sap from the leafy twigs is used to treat chest pain, side pain and
33
worms, and fever, root extract from the plant is drunk as a remedy (Matu, 2008).
The maceration of the crushed leaves of Clutia abyssinica given orally has been
2014). In Eastern Africa, soup from boiled roots is taken as a remedy for
Clutia abyssinica is used to treat yellow fever, malaria and infections of the ear,
nose and throat (Njoroge and Bushman, 2006). In Marakwet community, boiled
root extract is used to treat erectile dysfunction and works synergistically with
CHAPTER THREE
Fresh roots of Clutia abyssinica were collected randomly from Kaptebee village,
Turbo sub-county in Uasin Gishu county Kenya with the help of local herbalist
under accepted bio-conservation methods from its natural habitat (Appendix IV).
This process was conducted during the months of January-March 2016, a season
in which the local herbalist believed that the medicinal plant had its maximum
taxonomist for botanical authentication and a sample voucher was deposited at the
National Museums of Kenya herbarium Nairobi. Sample roots were then cleaned
with tap water to remove any dirt and chopped into smaller pieces. The root
samples of Clutia abyssinica were completely air dried under shade for two
weeks. The samples were then packed into burlap sacks and transported to
3.2 Extraction
uniformly mix the sample within the first 10 hours and left to stand for 48 hours.
Filtration was performed using Whatman‟s filter paper No.1 (sigma-aldrich). The
35
41°C under reduced pressure. The concentrate was kept in sealed containers at
Swiss albino mice aged between 5-6 weeks, weighing between 18-25 g and
Wistar albino rats aged 8-9 weeks and weighing 120-140 g were used in this
study. All the experimental animals were housed in the animal house at Kenyatta
throughout the entire experimental period. The animals were fed on rodent pellet
diet and water ad libitum. Wistar albino rats were utilized in testing for antipyretic
activity assay while for analgesic and anti-inflammatory activities assays, Swiss
albino mice were used. In this study ethical guidelines and procedures were
described by Singh and Majumdar (1995) and Akuodor et al. (2011). The
experiment animals were grouped into 6 groups of five animals each. Prior to pain
animals were fasted for 12 hours but were allowed access to water ad libitium.
Pain was induced by injecting 3% acetic acid solution at a dose of 20 ml/kg body
weight into the left side of the abdomen intraperitoneally. Immediately after
turning of body trunk of the laboratory animal was seen as an indication of pain.
The different groups were treated as follows; Group I (Normal control) was
administered with 10% dimethyl sulphoxide (DMSO) only but pain was not
induced. All the animals in group II (negative control) were induced with pain and
(positive control) were induced with pain and administered with the reference
drug (diclofenac 15 mg/kg body weight). Group IV-VI were induced with pain
and administered with DCM root extract of C. abyssinica at dose levels of 50, 100
and 150 mg/kg body weight respectively. This design is summarized in table 3.1.
Group Treatment
I (Normal control) 10% DMSO only
II (Negative control) 3 % Acetic acid
III (Positive control) 3 % Acetic acid + diclofenac (15
mg/kg bw)
IV (Experimental group A) 3 % Acetic acid + C.abyssinica (50
mg/kg bw)
V (Experimental group B) 3 % Acetic acid + C.abyssinica (100
mg/kg bw)
VI (Experimental group C) 3 % Acetic acid + C.abyssinica (150
mg/kg bw)
37
Thirty minutes after different treatments were administered, each mouse in groups
II−VI was intraperitoneally (ip) injected with 3% acetic acid into the left side of
the abdomen at a dose of 20 ml/kg body weight to induce pain sensation except
those in group I, which were administered with the vehicle (10% DMSO) only.
Each mouse was then placed in a transparent observation box and the number of
abdominal constrictions (writhes) for each mouse was counted for 30 minutes
percentage writhing inhibition was then calculated using the formula described by
Where;
Male Wistar albino rats were grouped into 6 groups of five rats each. Prior to
fever induction, rectal temperature of all the experimental animals were measured
using digital thermometer and recorded. All the experimental animals were fasted
doses but were allowed access to water ad libitium. Turpentine solution was used
weight and the animals left for one hour (Kuochung et al., 2006). A raise in rectal
temperature of Wistar albino rats by 0.8°C after one hour was termed pyretic and
One hour after fever induction, pyretic animals in groups IV-VI were
administered intraperitoneally with the three experimental doses of 50, 100 and
150 mg/kg body weight respectively. Pyretic animals in group III (Positive
control) were administered with aspirin (100 mg/kg body weight). Pyretic animals
Experimental animals in group I (normal control) were not induced with fever but
Group Treatment
I (Normal control) DMSO (10%)
II (Negative control) Turpentine (20%)
III (Positive control) Turpentine (20%) + Aspirin (100 mg/kg
bw)
IV (Experimental group A) Turpentine (20%) + C.abyssinica (50
mg/kg bw)
V (Experimental group B) Turpentine (20%) + C.abyssinica (100
mg/kg bw)
VI (Experimental group C) Turpentine (20%) + C.abyssinica (150
mg/kg bw)
Plant extract doses were prepared on the same day of experiment. All the
were measured by inserting a well lubricated digital thermometer about 3cm into
the anus of the rats. The mean body temperature of Wistar albino rats was
recorded at 20 minutes intervals over 1 hour before turpentine injection and was
recorded at an interval of one hour continuously for four hours after treatments.
Where,
the sub-plantar region of right hind paw of the Swiss albino mice according to
procedure described by Winter et al. (1962). Swiss albino mice (male) were
grouped into 6 groups of 5 animals (n=5) and treated as follows; each mouse in
group I (normal group) was administered with the vehicle (10% DMSO) only but
inflammation was not induced. All the mice in Group II (negative control) were
induced with inflammation and administered with the vehicle (10% DMSO) while
40
those in group III (positive control) were induced with inflammation and
administered with diclofenac (15 mg/kg body weight) one hour prior to
hind paw of Swiss albino mice. Animals in groups IV-VI were induced with
inflammation and administered with the plant extract at 50 , 100 and 150 mg/kg
Group Treatment
I (Normal control) DMSO (10%)
II (Negative control) Carragenaan (1%)
III (Positive control) Carrageenan (1%) + Diclofenac (15
mg/kg bw)
IV (Experimental group A) Carrageenan (1%) + C. abyssinica (50
mg/kg bw)
V (Experimental group B) Carrageenan (1%) + C.abyssinica (100
mg/kg bw)
VI (Experimental group C) Carrageenan (1%) + C.abyssinica (150
mg/kg bw)
Measurement of the hind paw diameter was carried out using a vernier calliper in
order to find out the diameter of right hind paw immediately before and after 1, 2,
inhibition was calculated according to the formula described by Jia et al. (2003);
41
Where;
inflammatory activities.
3.7.1 Flavonoids
A quantity of 1 ml of the plant root extract was put in a test tube followed by
Dry crude extract of the plant (5 mg) was dissolved in 2 ml chloroform and 1 ml
of acetic acid added to it. Concentrated (1ml) sulphuric acid was carefully added
42
to the solution alongside the test tube. Formation of a reddish violet colour
3.7.3 Steroids
To 0.5g of the extract, 2ml of glacial acetic acid containing four drops of 10%
ferric chloride (FeCl3) solution was added and under-layered with 1ml of
3.7.5 Phenolics
A solution of 2ml of the extract was put in a test tube and 1ml of ferric chloride
solution added carefully. Formation of blue to the green color indicated the
presence of phenolics.
added along the sides of test tube. Appearance of white creamy precipitate
shaken vigorously for 15 minutes. A foam layer was formed at the top of the
In this study, quantitative and qualitative data was obtained and recorded.
diameter and the rectal temperature were obtained from experimental animals,
recorded and tabulated on a broad sheet using Ms Excel program. Data obtained
was then exported to statistical software (Minitab version 17.0) for analysis. Data
Standard Error of the Mean (SEM). Results among the groups was analysed for
statistical significance using one way ANOVA followed Tukey‟s post hoc test for
significant.
44
CHAPTER FOUR
RESULTS
extracts of C. abyssinica into Swiss albino mice at all the three dose levels
(50, 100 and 150 mg/kg body weights), reduced the number of abdominal
(P> 0.05; Table 4.1). The DCM root extract of C. abyssinica, at the three
dose levels (50, 100 and 150 mg/kg body weight), reduced the number of
temperature (Table 4.2; Figure 4.1). In the first hour after treatment with the
experimental doses, all the three doses (50, 100 and 150 mg/kg body weight),
lowered the elevated anal temperature by 0.81%, 0.68% and 1.14% respectively
(Table 4.2; Figure 4.1). On the other hand, aspirin lowered the anal temperature
significantly by 3.32%, indicating a stronger antipyretic activity than the all the
three experimental doses (P˂ 0.05; Table 4.2; Figure 4.1). However, the
antipyretic effects of all the three doses of the DCM root extract of C.abyssinica
In the 2nd hour, experimental animals administered with DCM root extract of C.
abyssinica at 50, 100 and 150 mg/kg body weight, reduced the anal temperature
by 2.54%, 1.84% and 2.48% respectively. The reference drug, at this hour,
reduced rectal temperature by 3.53% (Table 4.3; Figure 4.1). At this hour, a dose
of 50 mg/kg body weight demonstrated the highest fever reducing activity. Even
though the three experimental doses of C. abyssinica and the reference drug
In the third hour, the DCM root extract of C. abyssinica, at the three doses
(aspirin) reduced elevated rectal temperature by 4.56% (Table 4.2; Figure 4.1).
The antipyretic effects of 150 mg/kg body weight and 100 mg/kg body weight
were comparable to the reference drug (aspirin) (P> 0.5; Table 4.2).
At the end of four hours of the test period, the DCM root extract of C. abyssinica
at all three dose levels, reduced the rectal temperature by 2.44%, 2.88% and
the experimental groups at all the three dose levels (50, 100 and 150 mg/kg body
(P> 0.5; Table 4.2; Figure 4.1). The antipyretic effects of 100 mg/kg body weight
and 150 mg/kg body weight were comparable to the reference drug (aspirin) (P>
Table 4. 2: Antipyretic activities of DCM root extracts of C. abyssinica on Turpentine-Induced pyrexia in Wistar albino rats
Group Treatment % change in anal temperature in (°C) after dose
administration
0hr 1hr 2hr 3hr 4hr
Generally, the DCM root extract, at the three doses demonstrated an anti-
mice. This was indicated by reduction in the diameter of the hind paw
after administration of the three extract doses (Table 4.3 and Figure 4.2).
In the first hour, the DCM root extract of C. abyssinica, at 50, 100 and 150
mg/kg body weight, reduced the inflamed hind paw diameter by 0.88%,
inflammatory effects of 50, 150 mg/kg body weight and diclofenac were
In the 2nd hour, the DCM root extract of C. abyssinica, at 50, 100 and 150
mg/kg body weight and diclofenac reduced the inflamed hind paw
anti-inflammatory effects of 100 mg/kg and 150 mg/kg body weight were
activities of the other two dose levels (100 mg/kg body weight and 150
In the 3rd hour, the DCM root extract at 50, 100 and 150 mg/kg body
weight and diclofenac reduced the inflamed hind paw diameter by 1.61%,
3.59%, 4.64% and 5.01% respectively (Table 4.3; Figure 4.2). The anti-
inflammatory effects of 100 mg/kg body weight and 150 mg/kg body
weight were not significantly different (P>0.05; Table 4.3; Figure 4.2).
In the fourth hour, the DCM root extract at 50, 100 and 150 mg/kg body
weight reduced inflamed hind paw diameter by 1.84%, 4.30% and 5.34%
activities of 150 mg/kg body weight and 100 mg/kg body weight were not
weight was significantly different from the other two doses and diclofenac
Table 4. 3: Anti-inflammatory activities of DCM root extracts of C. abyssinica on carragenaan-induced inflammation in Swiss albino
mice
The DCM root extract of C. abyssinica upon phytochemicals test revealed the
(Table 4.4).
CHAPTER FIVE
5.1 Discussion
Various injuries and diseases are most often presented with fever, pain and
inflammation among other clinical signs. Although during the past years there has
analgesics and glucocorticoids pose a lot of side effects after long term use like
cardiac abnormalities, peptic ulcers, prolonged bleeding, hepatic failure and renal
failure among others effects leading to their limited use in clinical settings
conventional drugs, search for newer drugs from medicinal plants with analgesic,
medicinal plants are an important option into discovery of novel drugs because
The present study was designed to evaluate the analgesic, antipyretic and anti-
To evaluate the analgesic activity of the DCM root extract, acetic acid-induced
pain test was used to induce abdominal writhings in Swiss albino mice. Acetic
acid-induced pain test has widely been used for screening new analgesic agents
The DCM root extract of C. abyssinica in this study, exhibited analgesic activities
Swiss albino mice after treatment with the extract. After thirty minutes of test
period, the DCM root extract at 50 mg/kg body weight demonstrated the highest
analgesic activity by reducing the number of writhes by 49.77%, while 100 mg/kg
body weight decreased the number of writhes by 38.6% and the dose of 150
mg/kg body weight decreased the number of writhes by 33.95% in a reverse dose
These findings strongly suggest that the DCM root extract of C. abyssinica posses
writhings in this study is in agreement with a study carried out by Mworia et al.
mice. The findings are also in line with studies by Kariuki et al. (2012) on
peripheral pain. The same effect in reduction of number of abdominal writhes was
observed by Elson et al. (2007) while examining the analgesic and anti-
mice models.
abyssinica, are in agreement with a study carried out by Gitahi et al. (2015) on
analgesic activities of root and leaf extract of Carissa edulis in Wistar albino rats.
The higher analgesic activity of a dose level of 50 mg/kg body weight than 150
mg/kg body weight (Table 4.1) might be attributed to the fact that when certain
limits in dose ranges are exceeded, the activity of that particular agent is reduced.
Also, high dose concentration of the extract could be taking longer time to diffuse
across peritoneum cavity in that compounds that are more concentrated takes
longer period to diffuse while those with lower concentration takes a shorter time.
drug, suggesting that the lower dose was more effective in managing pain induced
by acetic acid.
The dose range of (50, 100 and 150 mg/kg body weight) applied in bioscreening
the assays were comparable to dose ranges used by Tayyaba et al. (2015) while
activity of methanolic leaf extract of Kigelia africana (lam) benth and stem bark
extract of Acacia hockii de wild in Swiss albino mice. Likewise, Elson et al.
(2007) used a similar dose range while studying the analgesic and anti-
acetic acid-induced test, tail flick test and croton oil-induced ear edema test.
(Asfar et al., 2015; Kumar et al., 2015). An agent that lowers the number of
2009). Flavonoids have the ability to disrupt synthesis of eicosanoids (Robak and
1994). Besides flavonoids, alkaloids also have been associated with the ability to
inhibit pain perception (Uche et al., 2008). Alkaloids, for example berberine from
the extract might have antagonized peripheral mediators of pain and thereby
brewer‟s yeast and turpentine. These agents are considered exogenous pyrogens
(Petrova et al., 1978; Vasundra and Divya, 2013). Turpentine is a clear flammable
liquid with pungent odour and bitter taste, refined from resin pine. Turpentine
causes tissue damage and induces acute phase response as well as fever (Wieslaw
high fever pattern (Kuochung et al., 2006). Based on this information, turpentine
Figure 4.1). Findings from this study were comparable to other earlier research
animal models. Similar study carried out by Nthiga et al. (2016) showed that the
albino rats. Likewise, a study carried out by Kamau et al. (2016b) on antipyretic
properties methanolic leaf extracts of Kigelia africana (Lam) and Acacia hockii
The DCM root extract of C. abyssinica, lowered the elevated rectal temperature
more in the fourth hour than (in the third hour, the second and the first hour)
(Table 4.2; Figure 4.1). This might be attributed to the fact that most of the
bioactive compounds may have not been completely absorbed across the
peritoneum cavity while in the third and fourth hours most of the bioactive
62
compounds may have been absorbed across the peritoneum cavity thereby causing
higher antipyretic activities. Lower percentage inhibition in first and the second
hours could also be attributed to the fact that the drug needed time to be
antiviral drug (Jarko et al., 2008) . Another factor is that at certain dose ranges,
the active antipyretic phytochemical compounds in the extract were not sufficient
enough to lower the rectal temperature. However, the plant extract at 50 mg/kg
body weight lowered the rectal temperature in the second hour, then reducing
antipyretic activity in the third and fourth hours respectively (Table 4.2; Figure
4.1). This scenario could be ascribed to the fact that most of the bioactive
compounds might have been quickly metabolized and excreted because of the
In the fourth and the third hours, the DCM root extract of C. abyssinica, showed a
induced pyrexia in Wistar albino rats (Table 4.2; Figure 4.1). The dose dependent
response observed in this study are in agreement with studies by Mbiri et al.
another related study carried out by Tosan et al. (2014) on antipyretic activities of
From the data obtained in this study (table 2), the DCM root extract of C.
reference drug. The extract might have inhibited cyclooxygenase enzyme, a key
Moreover, the extract may have reduced the concentration of PGE2 in the
and Nwafor, 2010). However, other mechanism for blocking fever for example
glycosides and saponins (Table 4.4). Flavonoids have been found to inhibit
al., 2008). This study correlates with a study by Gitahi et al. (2015), that bioactive
64
is the most used primary test for screening new anti-inflammatory agents and
constitute a simple and routine animal model for evaluation of inflammation (Jain
et al., 2001; Paschapur et al., 2009). Carragenaan is obtained from a sea weed
injected into the hind paw leg of the rat/mice (John and Nodine, 1999). It is also
The DCM root extract of C. abyssinica in this study demonstrated strong anti-
by reducing the diameter of the edema (Table 4.3; Figure 4.2). There are some
correlations, between results obtained from this study and other studies on
Similar study conducted by Tukappa et al. (2015), on in vitro and in vivo anti-
initiating inflammation, signifying that the extract may have acted on cyclo-
dependent response was observed from the first hour to the fourth hour.
Reduction in the size of the hind paw edema was more in the fourth hour than in
the third, second and the first hours respectively. These differences in percentage
inhibition in the fourth hour and the first hour might be attributed to the fact that
absorption within first hour was slow but consistent. In the third and fourth there
inflammation more than in the first and the second hours. Lower percentage
inhibition in the first and the second hours might be ascribed to the fact that the
et al., 2008). The plant extract at 150 mg/kg body weight exhibited higher anti-
66
inflammatory effects than the other two dose levels in the entire experimental
compounds in the dose level than the other two doses. Another factor is small
concentration of active phytochemicals in the other two doses might have been
obscura in animal models. Likewise, a study carried out by Mbiri et al. (2016a)
metabolism (Chi et al., 2001; Jang et al., 2002). According to Vasudevan et al.
(2007), saponins have been shown to inhibit inflammation and therefore acting as
saponins present in the extract might have acted synergistically to bring forth anti-
inflammatory activities.
5.2 Conclusions
with the extract and the reference drug indicates that extract inhibited pain
mediators.
and the reference drug indicates that the extract reduced fever.
treatment with the extract and the reference drug indicates that the extract
5.3 Recommendations
agent.
69
for the activity which may lead to discovery of compounds that might be
inflammatory agents.
fever and inflammation. This will ensure that biomarkers responsible for
pain, fever and inflammation that are released during this process of pain,
iv. Evaluation of acute and chronic toxicity to determine the safety of the
REFERENCES
Adesokan, A.A., Yakubu M.T., Owoyele, B.V., Akanji, M.A., Soladoye, A.O.,
and Lawal, O. (2008).Effect of administration of aqueous and ethanol extracts of
Enantia chlorantha stem bark on brewer‟s yeast induced pyresis in rats. African
Journal of Biochemistry. 2: 165-169.
Afsar, T., Khan, M., Razak, S., Ullah, S., and Mirza, B. (2015). Antipyretic,
anti-inflammatory and analgesic activity of Acacia hydaspica R. Parker and its
phytochemical analysis. BMC Complementary and Alternative Medicine.15: 136-
145.
Akuodor, G.C., Anyalewechi, N.A., Udoh, F.V., Ikoro, N.C., Akpan, J.L.,
Gwotmut, M.D., Iwuanyanwu, T.C., and Osunkwo, U.A. (2011).
Pharmacological evaluation of Verbena hastate leaf extract in the relief of pain
and fever. Advances in Pharmacology and Toxicology. 3: 1–8.
Alam, M.A., Subhan, N.A., Awal, M., Alam, M.S., Sarder, M.L., and Sarker,
S.D. (2009). Antinociceptive and anti-inflammatory properties of Ruellia
tuberosa. Pharmaceutical Biology. 47, 209-214.
Apkarian, A.V., Bushnell, M.C., Treede, R.D., and Zubieta, J.K. (2005).
Human brain mechanisms of pain perception and regulation in health and disease.
European Journal of Pain. 9: 463-484.
Arome, D., Akapabio, I.S., Onalike, E.I., and Agbafor, A. (2016). Pain and
inflammation: Management by conventional and herbal therapy. Indian Journal of
Pain. 28: 5-12.
Beg, S., Swain, S., and Hasan, H. (2011). Systematic review of herbals as
potential anti-inflammatory agents: Recent advances, current clinical status and
future perspectives. Pharmacognosy Reviews. 5: 120–137.
Bhagyasri, Y., Lavakumar, V., Diva M., and Ashok, C. (2015). An overview
of anti-inflammatory activity of Indian medicinal plants: International Journal of
Research Pharmaceutical and Nano Science. 4: 1-9.
Blatteis, C.M. (2007). The onset of fever: new insights into its mechanism.
Progress in Brain Research. 162: 3–14.
Boursinos, L.A., Karachalios T., Poultisides L., and malizos, T.K. (2009). Do
steroids, conventional non-steroidal anti-inflammatory drugs and selective Cox-2
inhibitors adversely affect fracture healing. Journal of Musculoskelet Neuronal
Interact. 9: 44-52.
Cartmell, T., Poole, S., Turnbull, A.V., Rothwell, N. J., and Luheshi, G. N.
(2000). Circulating interleukin-6 mediates the febrile response to localized
inflammation in rats. Journal of Physiology. 525: 653-661.
72
Chi, Y., Jong, H., Son, K., Chang, H., Kang, S., and Kim, H. (2001). Effects of
naturally occurring prenylated flavnoids on enzymes metabolizing arachdonic
acid: cyclooxygenases and lipooxygenases. Biochemical Pharmacology. 62:1185-
1191.
Craig, C.R., and Stitzel, R.E. (2003). Modern Pharmacology with Clinical
Applications. Lippincott Williams & Wilkins, Philadelphia, 5th edition. pp 832.
Curtis, C.L., Harwood, JL., Dent, CM., and Caterson, B. (2004). Drug
Discovery Today. Journal of Drug Discovery and Therapeutics. 9: 165-174.
Dacie, J.V. (1958). Practical Hematology, 2nd edition. A Churchill Ltd., London.
pp. 38-48.
D'Armour, F.E., Blad, F.R., and Belden, D.A. (1965). Manual for Laboratory
work in Physiology, 3rd edition. The University of Chicago Press, Chicago.
Illinois. pp.4-6.
Dirosa, M., Giroud J.P., And Willoughby, D.A. (1971): Studies of the acute
inflammatory response induced in rats in different sites by carrageenan and
turpentine. Journal of Pathology. 104: 15-29.
Dubey, R.C., and Maheswari, D.K. (2005). A text book of Microbiology by and
5th edition published by S.Chand. New delhi pp 67.
Elisabetskey, E.A., Ahmador, RR., Nunse, D.S., and Carvallo, A.T. (1995).
Analgesic activity of Phshotua Colorata Muell. Journal of Ethnopharmacology.
48: 77–83.
Elson. A., Lucio, R., Itamar, S., Lecia, G., Gilmar., A and Luciano, M.
(2007). Anagesic and anti-inflammatory effects of cheiloclinium cognatum root
barks. Brazillian Journal of Pharmacognosy. 17: 508-513.
Ezeja, M.I., Ezeigbo, I.I., and Madubuike, K.G. ( 2011). Analgesic activity of
the methanolic seed extract of Buchholzia coriacea. Research Journal of
Pharmaceutical, Biological and Chemical Sciences. 2: 182-187.
Fitzgerald, G.A. (2004). Coxibs and cardiovascular disease. The New England
Journal of Medicine. 351: 1709–1711.
Friedman, R., and Hughes, A.L. (2002). Molecular evolution of the NF-κβ
signaling system. Immunogenetics. 53: 964–974.
Fujiyoshi, T., Hayashi, I., Ohishi, S., Kuwashima, M., Iida, H., Dozen, M.,
Taniguchi, N., Ikeda, K., and Ohnishi, H. (1989). Kaolin-induced writhing in
mice, a new model of possible bradykinin-induced pain for assessment of
analgesic agents. Agent and Actions. 27: 332-334.
Ghosh, S., May, M.J., and Kopp, E.B. (1998). NF-κβand Rel proteins:
evolutionarily conserved mediators of immune responses. Annual Review of
Immunology. 16: 225–231.
Gitahi, S.M., Maina, M.W., Njagi, J.M., Mworia, J.K., Juma, K.K., Umar,
A., Mwonjoria, K.J., Njoroge, W.A., Ngugi, M.P., and Mburu N.D. (2015).
Antipyretic Properties of Dichloromethane: Methanolic Leaf and Root Bark
74
Green, A.F., Young, P.A., and Godfrey, E.L. (1951). A comparison of heat and
pressure analgesimetric methods in rats. Britain pharmacology. 6: 572-585.
Gupta. M., Mazumder, U.K., Gomathi, P., and Selvan, V.T. (2006). Anti-
inflammatory activity of extract of Vernonia amygdalina. Complementary and
Alternative Medicine. 6: 36-43.
Guyton, A.C., and Hall, J.E. (2006). Textbook of Medical Physiology. 11th
edition. Elsevier Saunders, Philadelphia. pp 429-434.
Guyton, A.C., and Hall., J.E. (2000). Textbook of medical physiology. 10th
edition: Saunders Philadelphia. pp 822–823.
Hamalainen, M., Nieminen, R., Asmawi, M.Z., Vuorela, P., Vapaatalo, H.,
and Moilanen, E. (2011). Effects of flavonoids on prostaglandin E2 production.
Planta Medicina. 77: 1504-1511.
Inotai, A., Hanko, B., and Meszaro, A. (2010). Trends in the non-steroidal anti-
inflammatory drug market in six central-eastern European countries based on
retail information. Pharmacoepidemiology and Drug Safety. 19: 183–190.
Jain, N.K., Patil, C.S., Singh, A., and Kulkarni, S.K. (2001). A simple
technique to evaluate inflammatory pain along with anti-inflammatory studies in
carrageenan-induced paw edema. Indian Journal of Pharmacology. 33: 114-115.
Jang, D., Cuendet, M., Hawthorne ,M., Kardono, L., Kawanishi, K., and
Fong, H. (2002). Prenylated flavnoids of the leaves of Mucaranga conifera with
inhibitory activity against cyclooxygenase-2. Phytomedicine. 61: 867-872.
Jarko, R., Hanna, K., Tycho, H., Dooman, O., Tomi, J., and Jauko, S. (2008).
Prodrugs: Designs and Clinical Application. Nature Reviews Drug Discovery. 7:
255-270.
Jeruto, P., Charles, M., Lukhoba, C., and George, O. (2011). Phytochemical
constituents of some medicinal plants used by the Nandis of South Nandi district,
Kenya; Journal of Animal and Plant Sciences. 9: 1201–1210.
75
Jia, w., Gao, W.Y., Cui, N., and Xiao, P.G. (2003). Anti-inflammatory effects of
an herbal medicine (xuan-ju agent) on carragenaan and adjuvant-induced paw
edema in rats. Journal of Ethnopharmacology. 89: 139-141.
John, H., and Nodine, M.D. (1999). Year Book Medical. Publishers Inc;
Chicago. pp.492.
Kamau, J.K., Nthiga, P.M., Safari, V.C., Njagi, S.M., Mwonjoria, J.K.,
Ngerenwa, J.N., and Ngugi, M.P. (2016a) Anti-inflammatory activity of
methanolic leaf extract of Kigelia africana (Lam) benth and Acacia hockii de wild
in mice. Journal of Developing Drugs. 5: 158-166.
Kamau, J.K., Nthiga, P.M., Safari, V.C., Njagi, S.M., Mwonjoria, J.K.,
Ngugi, M.P., and Ngerenwa, J.N. (2016b). Antipyretic properties of methanol
stem bark extracts of Acacia hockii de wild and Kigelia africana (Lam) benth in
Wistar rats. Journal of Pharmacognosy and Natural Products. 2: 118-124.
Kariuki, H.N., Kanui, T.I., Yenesew, A., Patel, N.B., and Mbugua, M.P.
(2012). Antinociceptive activity of Toddalia asiatica (L) Lam in models of central
and pheripheral pain. Phytopharmacology. 3: 122-129.
Kelly, K.W., Bluthe, R.M., Dantzer, R., Zhou, J.H., Shen, W.H., Johnson,
R.W., and Broussard, S.R. (2003). Cytokine induced sickness behaviour . Brain
Behaviour and Immunology. 1: 2-8.
Kipkore, W., Benard, W., Hillary, R., and Gabriel, K. (2014). A study of
medicinal plants used by the Marakwet community in Kenya. Journal of
Ethnobiology and Etnomedicine. 10: 32–37.
Kluger, M.J. (1991). Fever: role of pyrogens and cryogens. Physiology Review.
71: 93–127.
Kokate, C.K., Purohit, A.P., and Gothale, S.B. (2002). Text book of
pharmacognosy 18th edition Nirali prakasan Ltd. Pune India.
Kokwaro, J.O. (2009). Medicinal plants of East Africa. 3rd edition. Kenya
literature bureau limited. Nairobi. pp 130.
Kou. J., Ni, Y., Li, N., Wang, J., Liu, L., and Jiang, Z.H. (2005). Analgesic and
anti-inflammatory activities of total extract and individual fractions of Chinese
medicinal ants Polyrhachis lamellidens. Biological and Pharmaceutical Bulletin.
28; 176–180
Kumar, A., Agarwal, K., Maurya, K.A., Shanker, K., Bushra, U., and
Tandon, S. (2015). Pharmacological and phytochemical evaluation of Ocimum
sanctum root extracts for its antiinflammatory, analgesic and antipyretic activities.
Pharmacognosy Magazine. 11: 17-24.
Kumar, S., Bajwa, B.S., Singh, K., and Kalia, A.N. (2013). Anti-inflammatory
activity of herbal plants. International Journal of Advances in Pharmacy, Biology
and Chemistry. 2: 77-88.
Kuochung, T., Haruko, F., Yasuo, Y., and Yuzo, T. (2006). Effects of
turpentine induced fever during the enamel formation of rat incisor. Archives of
oral biology. 51: 464-470.
Kupeli, E., Kosar, M., Yesilada, E., and Baser, K. (2002). A comparative study
on the antiinflammatory, antinociceptive and antipyretic effects of isoquinoline
alkaloids from roots of Turkish Berberis species. Life Sciences. 72: 645-652.
Lalenti, A., Ianaro, A., Moncada, S., and Di Rosa, M. (1995). Modulation of
acute Inflammation by endogenous nitric oxide. Europium Journal
Pharmacology. 211: 177-182.
Lange, C., Hemmrich, G., Klostermeier, U.C., Lopez, J.A., and Miller, D.J.
(2001). Defining the origins of the NOD-like receptor system at the base of
animal evolution. Journal of Molecular Biology and Evolution. 28: 687–702.
Lapah, P.T., Noa, P.A., and Ogbonna, O.J. (2014). Anti-Pyretic and Analgesic
Potentials of Aqueous Extract of Phragmanthera capitata S. Balle in Albino Rats.
American Journal of Pharmacy and Pharmaceutical Sciences.1: 37-43.
Lascelles, B.D., Court, M., Hardie, E.M., and Robertson, S.A. (2007).
Nonsteroidal anti-inflammatory drugs in cats: a review. Veterinary Anaesthesia
and Analgesia.34 :228–250
77
Luna, S.P., Basilio, A.C., Steagall, V.M., Machado, L.P., Moutinho, F.Q.,
Takahira, R.K., and Brandao, C.V. (2007). Evaluation of adverse effects of
long-term oral administration of carprofen, etodolac, flunixin meglumine,
ketoprofen, and meloxicam in dogs. American Journal of Veterinary Research.
2007; 68: 258-264.
Marzouk, B., Marzouk, Z., Haloui, E., Fenina, N., Bouraoui, A., and Aouni,
M. (2010). Screening of analgesic and anti-inflammatory activities of Citrullus
colocynthisfrom southern Tunisia. Jounal of Ethnopharmacology. 128: 15-19.
Matu, E.N. (2008). Clutia abyssinica Jaub and Spach. In: Schmelzer GH, Gurib-
Fakim A, Editors. Prota11 (1): Medicinal plants/Plantes médicinales PROTA,
Wageningen, Netherlands.
Maze, M., Hunter, J.C., and Gaeta, R.R. (2000). Conscious sedation and pain.
In Carruthers SG, Hoffman BB, Melmon KL, Nierenberg DW (eds.): Melmon
and Morrelli's Clinical Pharmacology. McGraw-Hill, NewYork, Fourth edition pp
200.
Mbiri , J.W., Sichangi, K., Kisangau, P., Wilton, M., and Ngugi, M.P.
(2016a). Anti-inflammatory properties of methanolic bark extract of Terminalia
brownii in Wistar albino rats. International Journal of Current Pharmaceutical
Research. 8: 1-5.
Mbiri, J.W., Sichangi, K., Kisangau, P., Wilton, M., and Ngugi, M.P. (2016b).
Anti-pyretic properties of methanolic bark extracts of Terminalia brownii in
Wistar albino rats. Journal of Pharmacognosy and Natural Products. 2: 121-126.
78
Mwangi, B.M., Gitahi, S.M., Njagi, J.M., Mworia, J.K., Aliyu, U., Njoroge,
W.A., Mwonjoria, K.J., Ngugi, M.P., and Mburu, N.D. (2015). Anti-
inflammatory properties of dichloromethane:methanolic leaf extracts of
Caesalpina volkensii and Maytenus obscura in animal models. International
journal of current pharmaceutical research. 7: 83-87.
Mwonjoria, J.K., Kariuki, H., and Waweru, F.N. (2011). The antinociceptive
antipyretic effects of Solanum incanum (Lin.) in animal models, International
Journal of Phytopharmaceuticals. 2: 22-26.
Mworia, J.K., Gitahi, S.M., Juma, K.K., Njagi, J.M., Mwangi, B.M., Aliyu,
U., Njoroge, W.A., Mwonjoria, K.J., Mawia, A.M., Nyamai, D.W., Ngugi,
M.P., and Ngeranwa, J.N. (2015) Antinociceptive Activities of Acetone Leaves
Extracts of Carissa Spinarum in Mice. Medicinal and Aromatic Plants. 10: 1- 4.
Naude, P.J., Cromarty, A.D., and Rensburg, C.E. (2010). Potassium humate
inhibits carrageenan-induced paw oedema and a graft-versus-host reaction in rats.
Inflammopharmacology. 18: 33–39.
Nthiga, P.M., Kamau, J.K., Safari, V.Z., Mwonjoria, J.K., Mburu, D.N., and
Ngugi, M.P (2016). Antipyretic Potential of Methanolic Stem Bark Extracts of
Harrisonia abyssinica Oliv and Landolphia buchananii (Hallier F.) Stapf in
Wistar Rats. Journal of Applied Pharmacy. 8: 227-233.
Onasanwo, S.A., Fabiyi, T.D., Oluwole L.K., and Olaleye, S.B. (2012).
Analgesic and anti-inflammatory properties of the leaf extracts of Anacardium
occidentalis in the laboratory rodents. Nigerian Journal of Physiological
Sciences. 27: 65-71.
Palladino, M.A., Bahjat, F.R., Theodorakis, E.A., and Moldawer, L.L. (2003).
Anti-TNF-α therapies: The next generation. Nature Reviews Drug Discovery. 2:
736-746.
Parle, M., and Yadav, M. (2013). Laboratory models for screening analgesics.
International Research Journal of Pharmacy. 4: 15-19.
Paschapur, M.S., Patil, M.B., Kumar, R., and Patil, S.R. (2009). Evaluation of
anti-inflammatory activity of ethanolic extract of Borassus flabellifer L. male
flowers (inflorescences) in experimental animals. Journal of Medicinal Plants
Research. 2: 49–54.
Petrova, L., Nikolova, M., Nikolov, R., and Stefanova, D. (1978). Dipyrone
and acetylsalicylic acid comparative pharmacological research. Antipyretic, anti-
inflammatory and analgesic action. In: Ovtcharov R, Pola W (eds) Proceedings
Dipyrone. Moscow Symposium, Schattauer-Verlag, Stuttgart New York, pp 99-
107
Proell, M., Riedel, S.J., Fritz, J.H., Rojas, A.M., and Schwarzenbacher, R.
(2008). The Nod-like receptor (NLR) family: A tale of similarities and
differences. PLoS One Journal Impact. 3: 2199-2208.
80
Rang, H.P., Dale, M.M., Ritter, J.M., and Flower, M.C. (2006). Pharmacology,
7th edition, Churchill Livingstone. pp. 202-210.
Rang, H.P., Dale, M.M., Ritter, J.M., and Moore, P.K. (2003). Pharmacology.
Churchill Livingstone, Edinburgh, 5th edition . pp. 797.
Ravi, V., Saleem, T.S., Patel, S.S., Raamamurthy, J., and Gauthamam, K.
(2009). Anti-Inflammatory Effect of Methanolic Extract of Solanum nigrum Linn
Berries. International Journal of Applied Research in Natural Products. 2: 33-36.
Rehman, Q., and Sack, K.E. (1999). When to try COX-2-specific inhibitors:
Safer than standard NSAIDS in some situations. Postgraduate Medical Journal.
106: 95–106.
Richard, F., Michelle, A.C., and Luigi, X.C. (2008). Lippincott's Illustrated
Reviews. Pharmacology. Lippincott Williams & Wilkins. Philadelphia, Fourth
edition. 564.
Saper, C.B., and Breder, C.D. (1994). The neurologic basis of fever. New
England Journal of Medicine. 330: 1880–1886.
81
Semwal, D.K., Badoni, R., Semwal, R., Kothiyal, S.K., Singh, G.J., and
Rawat, U. (2010). The genus Stephania (Menispermaceae): Chemical and
pharmacological perspectives. Journal of Ethnopharmacology. 132: 369-383.
Sen, S., Chakraborty, R., and De, B. (2010). Analgesic and anti-inflammatory
herbs: A potential source of modern medicine. International Journal of
Pharmaceutical Sciences and Research. 11: 32–44.
Shah, B.S., Nayak, B.S., Seth, A.K., Jalalpure, S.S., Patel, K.N, Patel, M.A.,
and Mishra, A.D. (2006). Search for medicinal plants as a source of anti-
inflammatory and anti-arthritic agents. Pharmacognosy Magazine. 2: 77–86.
Smith, D.L., D’amour, M.C., and D’amour, F.E. (1943). The analgesic
properties of certain drugs and drug combinations. Journal Pharmacology
Experimental Therapeutics. 77: 184-193.
Sparkes, A., Heiene, R., Lascelles, B.D., Malik, R., Sampietro, L.R.,
Robertson. S., Scherk, M., and Taylor, P. (2010). NSAIDs and cats- it‟s been a
long journey. Journal of Feline Medicine and Surgery. 12 :519–538
Sund- Levander, M., Forsberg, C., and Wahren, L.K. (2002). Normal oral,
rectal, tympanic and axillary body temperature in adult men and women: a
systematic literature review. Journal of Caring Science. 16: 122- 129.
Tayyaba, A., Muhamed, R.K., Suhail, R., Shafi, U., and Bushra, M. (2015).
Antipyretic, anti-inflammatory and analgesic of Acacia hydaspica r parker and its
phytochemicals analysis. Complementary and Alternative Medicine. 15: 136-147.
Thorp, C.M. (2008). Pharmacology for the Health Care Professions. A John
Wiley & Sons, Ltd. Lodon; pp 243.
Tordera, M., Ferrandiz, M.L., and Alcaraz, M.J. (1994). Influence of anti-
inflammatory flavonoids on degranulation and arachidonic acid release in rat
neutrophils. Journal of Biosciences. 49: 235-240.
Tosan, C.A., Adeoti, O.K., Ojewuyi, O.B., and Olayemi, J. (2014). Antipyretic
activity of Abutilon mauritianum (jacq) roots in Wistar rats. International Journal
of Pharma Sciences and Research. 5: 75-81.
Tukappa, N.K., Ramesh, L.J., Sanjeev, C.B., and Hanumantapa, B.N. (2015).
Evaluation of in vitro and in vitro anti-inflammatory and toxicity studies of
methanolic extract of Rumex vesicarius Linn. Orient Pharmaceuticals Experiment
Medicine. 9: 35-43.
Trease, G.E., and Evans, M.C. (1983). Text book of Pharmacognosy. 12th
edition. Balliere, Tindall: London. Pp. 343-383.
Vane, J.R., and Bolting, R.M. (1995). New insights into the mode of action of
anti-inflammatory drugs. Journal Inflammation Research. 44: 1-10
Vane, J.R., Bakhle, Y.S. and Bolting, R.M. (1998). Cyclooxygenase-1 and 2.
Annual Review of Pharmacology and Toxicology. 38: 97-120.
Vasundra, D.P., and Divya, P.S. (2013). Antipyretic Activity of Ethanol and
Aqueous Extract of Root of Asparagus racemosus in Yeast Induced Pyrexia.
Asian Journal of Pharmaceutical Clinical Researh. 6; 190-19
Winter, C.A., and Poter, C.C. (1957). Effect of alterations in the side chain
upon anti-iflammatory and liver glycogen activities of hydrocortisone esters.
Journal of American Pharmaceutical Association Science. 46: 343-383.
Winter, C.A, Risley, E.A and Nuss, G.W. (1962). Carrageenan-induced oedema
in the hind paw of rats as an assay for anti-inflammatory activity. Proceeding of
Society for Experimental Biology and Medicine.111: 544-547.
Wieslaw, K., Mathew, J.K., Dariusz, S., Carole, C., Karin, R., Lisa, T., and
Hui, Z. (1998). IL-6 and IL-1β in fever. Studies using cytokine-deficient
(knockout) mice. Annals of the New York Academy of Sciences. 856: 33-47.
Young, A., Jordan, F., Ledingham, M., Thomson, A., Norman, J., and Greer,
I. (2002). Quantification of pro-inflammatory cytokines in myometrium, cervix
and fetal membranes during human parturition. Journal of the Society of
Gynecologic Investigation. 9: 133–137.
84
Young, P., and Saxena, M. (2014). Fever management in intensive care patients
with infections: Intensive Care. 18: 206–213.
85
APPENDICES
APPENDIX II: Effects of DCM root extract of C. abyssinica on turpentine-Induced pyrexia in Wistar albino rats
Positive Control Turpentine + Aspirin (100 mg/kg 39.14±0.35a 37.84±0.34b 37.76±0.34b 37.36±0.41b 37.2±0.42b
bw)
Experimental Turpentine + C.abyssinica (50 38.52±0.48ab 38.2±0.37ab 37.54±0.34b 37.6±0.49b 37.58±0.48ab
group A mg/kg bw)
Experimental Turpentine + C.abyssinica (100 38.08±0.48ab 37.82±0.32b 37.38±0.29b 36.98±0.31b 36.98±0.33b
group B mg/kg bw)
Experimental Turpentine + C.abyssinica (150 38.72±0.08a 38.28±0.11ab 37.76±0.16b 37.42±0.18b 37.48±0.20b
group C mg/kg bw)
87
APPENDIX III: Effects of DCM root extract of C. abyssinica on carrageenan-Induced inflammation in Swiss albino
………………….mice
APPENDIX IV: Source of the plant Turbo division. Courtesy of Google Earth maps
89
Model Summary
Means
C1 N Mean Grouping
Normal control 5 100.0 A
50 mg/kg 5 49.77 B
Positive control 5 46.51 B
100 mg/kg 5 38.60 B
150 mg/kg 5 33.95 B
Negative control 5 0.000000 C
90
Model Summary
Means
C1 N Mean Grouping
Positive control 5 3.320 A
150 mg/kg 5 1.137 B
50 mg/kg 5 0.812 B
100 mg/kg 5 0.680 B
Normal control 5 0.211 B C
Negative control 5 -1.077 C
Analysis of Variance
Model Summary
Means
C1 N Mean Grouping
Positive control 5 3.526 A
50 mg/kg 5 2.525 A
150 mg/kg 5 2.480 A
100 mg/kg 5 1.829 A B
Normal control 5 0.215 B C
Negative control 5 -1.392 C
Model Summary
Means
C1 N Mean Grouping
Positive control 5 4.553 A
93
Model Summary
Means
C1 N Mean Grouping
Positive control 5 4.964 A
150 mg/kg 5 3.203 A B
100 mg/kg 5 2.879 A B
50 mg/kg 5 2.436 B
Normal control 5 0.054 C
Negative control 5 -0.610 C
Method
Factor Information
Analysis of Variance
Model Summary
95
Means
C1 N Mean Grouping
150 mg/kg 5 3.300 A
Positive control 5 2.2090 B
100 mg/kg 5 1.8774 B
50 mg/kg 5 0.884 C
Normal control 5 0.0816 D
Negative control 5 -2.401 E
Model Summary
Means
C1 N Mean Grouping
150 mg/kg 5 3.761 A
Positive control 5 3.567 A
100 mg/kg 5 3.036 A
50 mg/kg 5 1.470 B
Normal control 5 0.0778 C
Negative control 5 -3.464 D
Model Summary
Means
C1 N Mean Grouping
Positive control 5 5.011 A
150 mg/kg 5 4.642 A B
100 mg/kg 5 3.586 B
50 mg/kg 5 1.608 C
Normal control 5 0.1621 D
Negative control 5 -4.348 E
97
Model Summary
Means
C1 N Mean Grouping
150 mg/kg 5 5.359 A
Positive control 5 5.356 A
100 mg/kg 5 4.305 A
50 mg/kg 5 1.839 B
Normal control 5 0.0806 C
Negative control 5 -4.735 D