Al-Mahbashi et al.
Universal Journal of Pharmaceutical Research 2020; 5(5):61-66
Available online on 15.11.2020 at http://ujpr.org
Universal Journal of Pharmaceutical Research
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Volume 5, Issue 5, 2020
RESEARCH ARTICLE
PHYTOCHEMICAL, ANTI-INFLAMMATORY, ANALGESIC, ANTIPYRETIC
AND ACUTE TOXICITY OF PSIADIA PUNCTULATA GROWING IN YEMEN
Hassan Al-Mahbashi1* , Bushra Abdulkarim Moharram2 , Tareq Al-Maqtari3,4
1
Department of Forensic Medicine & Clinical Toxicology, Faculty of Medicine and Health Sciences, University of Sana’a., Yemen.
2
Department of Pharmacognosy, Faculty of Pharmacy, University of Sana’a, Sana'a, Yemen.
3
Department of Pharmacology, Faculty of Pharmacy, University of Sana’a, Sana'a, Yemen.
4
Department of Pharmacology, Faculty of Pharmacy, International University for science and Technology, Damascus, Syria.
ABSTRACT
Background: Psiadia punctulata growing in Yemen is used traditionally for different medicinal purpose, such as in casts of
broken bones and for relief of pain, fever and abdominal pain.
Objective: To determine the chemical composition and to assess the anti-inflammatory, analgesic, and antipyretic activity of P.
punctulata leaf extracts.
Method: Phytochemical screening of P. punctulata ethyl acetate and ethanol extracts were performed using chemical tests and
thin layer chromatography. An acute oral toxicity study was conducted in rats by administering oral ethanol leaf extract of up to
5000 mg/kg. The anti-inflammatory activity of orally administered ethyl acetate and ethanol extracts (200, 400 mg/kg) and
diclofenac (20 mg/kg) were evaluated using a formalin-induced inflammation rat model. The analgesic activity of orally
administered ethyl acetate and ethanol leaf extracts (100, 200, 300mg/kg), compared to diclofenac (20 mg/kg) were evaluated by a
formalin-based test as well. The antipyretic activity of oral ethyl acetate and ethanol extracts (400 mg/kg) versus paracetamol
(150mg/kg) was assessed in Baker’s Yeast-induced pyrexia rats.
Results: The phytochemical analyses indicated the presence of alkaloids, carbohydrates, steroids, phenolic compounds/tannins,
phytosterols, saponins, gum and mucilage. The ethanol extract of the plant was apparently safe in rats at doses as high as 50 00
mg/kg body weight. Time- and dose-dependent anti-inflammatory activity of the ethyl acetate and ethanol extracts (200 and 400
mg/kg) were clearly observed in rats. The results showed that both extracts exerted significant analgesic and antipyretic effects.
Conclusion: Psiadia punctulata possess anti-inflammatory, analgesic and antipyretic activities with a wide safety margin.
Keywords: Analgesic, antipyretic, anti-inflammatory, Psiadia punctulata, Yemen.
Article Info: Received 22 September 2020; Revised 7 October; Accepted 26 October, Available online 15 November 2020
Cite this articleAl-Mahbashi H, Moharram BA, Al-Maqtari T. Phytochemical, anti-inflammatory, analgesic, antipyretic and
acute toxicity of Psiadia punctulata growing in yemen. Universal Journal of Pharmaceutical Research 2020;
5(5):61-66.
DOI: https://doi.org/10.22270/ujpr.v5i5.489
Address for Correspondence:
Dr. Hassan Al-Mahbashi, Department of Forensic Medicine and Clinical Toxicology, Faculty of Medicine and Health Sciences,
University of Sana’a, Sana’a, Yemen. Tel: 00967771703099; E-mail: hassanpharmad@gmail.com
hepatocellular carcinoma, bladder carcinoma and
carcinoma of the pharynx. Antioxidant, antifungal,
anti-Leishmanial and anti-malarial activities were also
observed 5,6. The plant is traditionally used for different
medicinal purpose in the Arab Peninsula. It is used by
locals in casts of fractured bones. Also, the leaf and
stem extracts are used to relieve pain and to speed
recovery in foot injuries of villagers who often walk
around barefooted. In east Africa (particularly in
Kenya), leaf decoction offers several benefits including
common cold and fever management and for protecting
cattle against ectoparasites1.The plant is also used for
its analgesic activity, particularly for abdominal pain 7.
In Yemen, the species were found in Taiz, Sumara,
INTRODUCTION
Psiadia punctulata belongs to the family Asteraceae, is
found in several African countries as well as in
Yemen1. The plant Psiadia Jacq. contain several
species2, three of which are found in Yemen including
Psiadia incanao, Psaidia punctulata and Psiadia
schweinfurthii3. Phytochemical studies of leaf exudate
of P. punctulata showed presence of flavonoid,
kaurenes and trachylobanediterpenes1,4. Several studies
have reported several biological activities for P.
punctulata. For instance, it has been shown to exert
cytotoxic activity against multiple types of cancer cell
lines including breast cancer, cervix cancer,
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Dhamar, Adhale, Hajja, Ibb, Shabwa and Hadramout.
It is mostly added there to casts to speed up recovery of
bones. The aim of this study is to carry out the
phytochemical screening of ethyl acetate and ethanol
fractions of the leaf extract of P. punctulata in addition
to assessing the anti-inflammatory, analgesic and
antipyretic activity of both extracts. The oral acute
toxicity of the ethanol leaf extract was also assessed in
vivo.
Economic Co-operation and Development (OECD)
were followed10. In brief, 36 rats (6 animals in 6
groups) were given only water for 16 hours. The
animals were then administered oral methanolic plant
extracts in Tween 80 (1% w/v) at serial concentrations
of 100, 1000, 2500, 4000 and 5000 mg/kg of body
weight whereas the control group were fed the vehicle
only. Several parameters were then monitored for 14
days11 including physical signs (weight, physical
appearance, eyes, mucous membranes, and fur/skin
condition), neurological abnormality (behavior,
tremors, diarrhea, salivation, seizures, and physical
activity) and mortality. A large dose of sodium
thiopentone (100 mg/kg intraperitoneal) was used to
euthanize the animals at the end of experiment12.
Evaluation of anti-inflammatory activity
Formalin-induced inflammation: This test was
performed as described previously9,13.Six groups (n =
6) of albino rats were used. All groups were injected
with 2% freshly prepared formalin (10 μl) into the sub
plantar region of right hind paw to induce
inflammation. One hour prior to inflammation
induction group 1was administered 10 ml/kg water p.o.
(vehicle), group 2 was administered 20 mg/kg
diclofenac sodium p.o., groups 3 and 4 were fed ethyl
acetate extracts 200 and 400 mg/kg p.o (respectively)
and groups 5 and 6 were given ethanol extracts 200 and
400 mg/kg p.o. (respectively).With a Vernier caliper,
paw volume was measured prior to administration of
inflammatory agent and then at predetermined time
points (1, 2, 3 and 4 hours after formalin injection).
Anti-inflammatory activity of the extract was evaluated
through this equation:
MATERIALS AND METHODS
P. punctulata was collected from the district of Bani
Saifin Bani Moharam, Ibb city, Yemen in November
2017. The plant was identified by Dr. Abdul-Wali AlKhulaidi (working at the Public Authority for Research
and Agricultural Extension, Yemen). The specimen
voucher of the plant was deposited in the department of
pharmacology, Faculty of pharmacy, Sana'a University.
The voucher number is pp17. Methanol 99.8%
(Scharlae, Spain), ethyl acetate (HiMedia, India),
formic acid (Fluka, Switzerland), paracetamol and
sodium diclofenac (Shaphaco Pharmaceutical Ind.Yemen), 0.9% NaCl (SMSCO, Saudi Arabia), Tween
80 (UniChem, Beograd), and thiopental (Rotexmedica,
Germany). Solvents/chemicals used were of standard
analytical grade.
Extract preparation
The leaves of the plant were thoroughly cleaned then
cut into small pieces before weighing them. Then the
leaves were put in sufficient amount of ethyl acetate for
20 seconds to get an ethyl acetate extract then the
leaves was soaked in ethanol at room temperature for 3
days to get an ethanol extract8. Filtration of extracts
was performed filtered using Whatman No.1 filter
paper. After that, the solvent was evaporated with a
rotary evaporator in a water bath with temperature not
exceeding 45°C. The extracts were stored in airtight
containers at room temperature until time of use.
Phytochemical screening
Carbohydrates, alkaloids, fixed oils/fats, glycosides,
poly phenols, tannins, sterols, peptides/proteins,
saponins, gum and mucilage were screened in the ethyl
acetate and ethanolic extracts using a standard
phytochemical screening procedure as previously
described9.
Animals
Mature male Albino rats, weighing 150-250 g were
obtained from the animal house of the College of
Science (at Sana’a University). The animals were put
in individual cages with controlled light, temperature
and humidity (six rats per cage). The animals were
maintained on standard diet and tap water and put in a
colony room. A 12/12 hr light/dark cycle and a
temperature of 21±2°C were maintained before and
during the experimentation period. The rats were
acclimatized to the laboratory conditions for 48h before
experimentation. The experiments were approved by
the Institutional Ethical Committee, Faculty of
Medicine, Sana`a University (23/10/2017).
Acute oral toxicity
To investigate the acute toxicity profile of P.
punctulata, the guidelines of the Organization for
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The edema reduction was assessed using the following
formula:
Where;
V0 represents rat paw volume prior to administration
of formalin, Vt represents rat paw volume after
formalin injection at a given time.
Evaluation of the analgesic activity
Formalin test: Thirty two male albino rats were
allocated into 8 groups (n=4).Group 1 (negative
control) were given normal saline 10 ml/kg and groups
2, 3, 4 were treated with ethyl acetate extract at doses
of 100, 200, 300 mg/kg, respectively. Groups 5, 6, 7
were administered ethanol extract at doses of 100, 200,
300 mg/kg, respectively. Group 8 (positive control)
were treated with 20 mg/kg diclofenac sodium. Thirty
minutes later, the rats were injected with 0.05 ml
formalin 2.5% into the right hind paw, then placed
immediately in separate plastic cages before injected
paw licking time and frequency were recorded for 30
min13.
Antipyretic Test
Yeast-induced pyrexia model in rats: The test was
performed as described earlier 14,15. Four rat groups (n
= 5) were used. Group1 (negative control) were
administered 10 ml/kg normal saline whereasgroup2
(positive control) were treated with 150 mg/kg
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Universal Journal of Pharmaceutical Research 2020; 5(5):61-66
paracetamol and groups 3 and 4 were treated with 400
mg/kg of the ethyl acetate and ethanol extract of P.
punctulata, respectively (all orally). Before inducing
fever, baseline rat rectum temperatures for all rats were
recorded with a digital thermometer. For pyrexia
induction, subcutaneous injections of 20% w/v Baker’s
yeast suspension (10 ml/kg) were administered. Rectal
temperatures were then taken after 19 hours. After that,
normal saline, paracetamol, ethyl acetate, and ethanol
extract were administered orally only to the rats with a
0.6°C (or 1°F) increase in rectal temperature. Rectal
temperatures were again recorded in the first, second,
third and fourth hours following treatments.
Statistical analyses
Statistically Package for Social Sciences (SPSS)
version 11.5 was utilized for data analysis. Data are
presented as means±Standard deviations (SD),
Categorical variables were represented by frequencies
and percentages. Paired T-test was implemented to test
the significance of the differences between every two
groups. Significance level was set at 0.05 and 0.01.
Table 1: Phytochemical screening of the ethyl acetate and ethanol extracts of P. punctulata
Phytochemical screening
Alkaloids
Carbohydrates
Fixed oils/ fats
Steroids
Anthraquinones
Phenolic
compounds/tannins
Phytosterols
Proteins
Saponins
Gum and Mucilage
Mayer’s test
Wagner’s test
Benedict’s test
Saponification
Salkowsk’s test
Borntrager’s test
Ferric chloride test
Lead acetate test
Mg and HCl reduction test
Libermann-Burchard’s test
Biuret test
Foam test
Ethyl acetate
extract
+
+
+
+
+
+
+
+
+
+
+
Ethanol
extract
+
+
+
+
+
+
+
+
+
+
+
+ = presence, - = absence, Mg = Magnesium, HCl = Hydrochloric acid.
Table 2: Anti-inflammatory activities of the ethyl acetate and ethanolic extract of P punctulata leaves and
diclofenac on formalin-induced edema in the right hind-limb of rats
Time
(hr)
0
1
2
3
4
Treatment
Control
3.01±0.040
3.91±0.20*
4.13±0.21*
4.13±0.21*
4.76±0.45*
Ethyl acetate extract (mm)
(% of inhibition )
200
400
2.98±0.09
3.23±0.20
3.56±0.33*
3.78±0.22*
3.25±0.28 (9%) 3.46±0.20 (8%)
3.21±0.31(10%) 3.25±0.19" (14%)
3.03±0.08"(15%) 3.25±0.19" (14%)
Ethanolic extract (mm)
(% of inhibition)
200
400
2.93±0.08
2.98±0.07
3.33±0.22*
3.33±0.18*
3.16±0.10* (5%)
3.15±0.15 (5%)
3.08±0.07" (8%)
3.05±0.05" (8%)
2.98±0.04" (11%) 3.00±0.06" (10%)
* = P≤ 0.05 compared to value at zero time," = P≤ 0.05 compared to value at 1 hour time
RESULTS
Anti-inflammatory Activity
Injecting rats with 0.1 ml 2% formalin into the right
hand foot pad resulted in a local inflammatory reaction
and edema. The size of edema increased gradually with
time following the injection compared to zero time in
group one and reached a maximum after 4 hours of
injection (table 2). Prior administration of diclofenac
(positive control) led to a marked decrease in the
inflammation and in the size of edema compared to
time zero.
The anti-inflammatory activity of
diclofenac started within 2 hours, and the effect peaked
after 4 hours post injection resulting in a 12% reduction
in edema. The results showed that the ethyl acetate
extract (200, 400 mg/kg b.w.) possessed antiinflammatory activity. At 4 hours post injection, the
two tested doses decreased the size of edema
significantly (15% and 14% reduction respectively)
compared to the edema size at 1 hour. Additionally, the
ethanol extract (200 and 400 mg/kg) resulted in less
anti-inflammatory effect and inhibited edema by 11%
and 10%, respectively.
Analgesic Activity
Formalin-induced pain in the right hind-limb of rats
was utilized to evaluate the analgesic activity of three
Phytochemical screening
The findings of the phytochemical analysis for the P.
punctulata ethyl acetate, and ethanol extracts are
illustrated in Table1. Bothextracts were positive for
alkaloids, carbohydrates, steroids, poly phenols,
tannins, phytosterols, saponins, gum and mucilage.
Acute toxicity test
The findings of the study indicate the safety of P.
punctulata methanolic extracts on rats. Even at high
doses of up to 5000 mg/kg, no apparent adverse
reactions or toxicity were noted in rats. No physical,
neurological, psychological abnormalities were
recorded during the 2 weeks period post oral extract
ingestion. The animal appeared physically active with
no alterations in appearance, skin/fur, salivation,
defecation, or sleeping patterns. Also, no behavioral
changes, neurological defects, comas or deaths were
observed. These data show the relative safety of P.
punctulata in living systems and indicate that the lethal
dose 50 (LD50) of P. punctulata methanolic extract in
rats is above 5000 mg/kg.
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Sodium
Diclofenac
20mg/kg
2.96±0.05
3.43±0.12*
3.16±0.16"(8%)
3.11±0.09"(9%)
3.01±0.04"(12%)
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doses of each extract at multiple doses (100, 200, 300
mg/kg). These extracts were compared with the
analgesic effect exerted by the powerful non-steroidal
anti-inflammatory drug diclofenac (20 mg/kg). Table 3
showed the extracts at different doses significantly
decreased both licking time and frequency compared to
those of the control group. Highest percent decrease in
licking time (57%) was achieved at 200 mg/kg dose of
ethanol extract. The percent reduction induced by
diclofenac sodium (20 mg/kg) was about 61%.
Interestingly, both extracts were also effective in
alleviating pain. For instance, the 300 mg/kg ethyl
acetate extract induced a 58% percent of reduction in
licking frequency while the 300 mg/kg ethanol extract
induced a 45% reduction. The analgesic activity
recordings showed that the reduction of liking time and
licking frequency by both extracts was generally not
dose-dependent.
Antipyretic Activity
As shown in Table 4, subcutaneous injections of
animals with 20 % w/v Baker’s yeast suspension (10
ml/kg) led to significant elevation in rectal
temperatures after 19 hrs. In group one (treated with
normal saline), rectal temperature continued to elevate
for 2 hours before decreasing in the 3rd and 4th hours
post normal saline treatment. In the positive group,
treated with the antipyretic drug paracetamol
(150mg/kg), the rectal temperature significantly
decreased in all selected time points compared to time
zero. Also, the rectal temperature of the group treated
with the ethyl acetate (400 mg /kg) extract significantly
decreased in all time points except in the 4th hour,
compared to the negative control group. On the other
hand, the rectal temperature of the group that was
given the ethanol extract (400 mg/kg), significantly
decreased in the first two hours but not in the 3rd or 4th
hours post treatment.
Table 3: The analgesic activity of the ethyl acetate and ethanolic extracts of P. punctulata leaves and diclofenac
on formalin-induced pain in the right hind-limb of rats
Treatment
Control
Ethyl acetate
extract
Ethanol
extract
Diclofenac
Dose
mg/kg
100
200
300
100
200
300
20
Licking time (sec)
(% of inhibition)
15.9±2.3
10.3±0.6* (35%)
8.5± 0.6* (47%)
9.9±0.9* (38%)
12.1±0.9* (24%)
6.9±0.1* (57%)
10.8±0.8* (32%)
6.2± 0.4* (61%)
Licking frequency/30min
(% of inhibition)
35.8±4.2
28.9±1.0* (19%)
23.5±3.1* (34%)
15±1.0* (58%)
20.2±4.6* (44%)
21±4.5* (41%)
19.8± 4.5* (45%)
24.8±1.2* (31%)
* = P≤ 0.05 compared to control
DISCUSSION
The present work established the anti-inflammatory
activity of P. punctulata leaves in vivo. Indeed, both
ethyl acetate and ethanol plant leaf extracts, at two
different doses of 200-400 mg/kg b.w., displayed
marked anti-inflammatory effect as shown by
decreasing size of formalin-induced rat paw edema in
rats. The paw edema reduction exhibited by the plant
extracts appeared dose-independent as using a 400
mg/kg concentration did not have a favorable antiinflammatory effect compared to that of using half that
concentration (200 mg/kg). For instance, four hours
post formalin injection, the edema reduction in the
groups treated with the 200 mg/kg and 400 mg/kg ethyl
acetate extracts were 15% and 14 % respectively.
Notably, the prominent reduction in edema and
inflammation by the extracts started after 2 hours
following formalin injection and continued throughout
the entire observation period of 4 hours, which
indicates their efficacy in alleviating the late phase of
inflammation if taken orally. The shown two-hour time
gap before the manifestation of the anti-inflammatory
activity of extracts are likely ascribed to the time
needed for the bioactive agents to distribute in body
fluids (and then distribute into target sites) or the time
needed for biotransformation inside the body to form
active metabolites endowed with the anti-inflammatory
activity. The anti-inflammatory effect of P. punctulata
is possibly mediated by inhibiting prostaglandins and
In this study, the phytochemical screening of P.
punctulataleaves as well as the biological activities of
the plant leaf extracts, including anti-inflammatory,
analgesic, antipyretic activities and acute toxicity were
investigated. The preliminary phytochemical analysis
of the ethanol and ethyl acetate leaf extracts indicated
the presence of chemical constituents which may
contribute to its claimed medicinal activities. The
chemicals detected using chemical test included
alkaloids, carbohydrates, steroids, bitters,poly phenols,
tannins, flavonoids, sterols, saponins, gum and
mucilage. To assess the anti-inflammatory activity
exerted by P. punctulata leaves, formalin-induced
inflammation rat model was utilized. This method is
known to predict the potential of a test agent to combat
acute inflammation by diminishing the action of the
inflammatory autocoids involved and has extensively
been used to evaluate the anti-inflammatory potential
of plant extracts in several studies16,17. When injecting
formalin in rat paws, a biphasic local inflammatory
response is induced. The first (early) phase is
associated with neurogenic pain whereas the second
(late) phase involves the activation of inflammatory
processes driven by the release of local mediators18.
Local inflammatory mediators produced during the late
phase of inflammation include prostaglandins,
serotonin, histamine, bradykinin and other cytokines19.
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prostaglandins that are responsible for the regular signs
other inflammatory mediators. However, more
of inflammation (redness, edema, itching etc)21.
investigations are needed to confirm that.
Inflammation is a natural biological response to insults
To determine the potential of P. punctulata to combat
and involves activation of various enzymes and local
pain, paw licking time and frequency were recorded
autacoids, cell migration, tissue breakdown and repair.
after injecting formalin in the right hind paw of rats.
Edema in an inflamed tissue occur due to increased
This test is used to demonstrate the involvement of
capillary permeability of water and albumin in plasma.
both central and peripheral pathways of analgesia and
Migration of white blood cells, particularly neutrophils
offers the advantages of mimicking clinical human
also occur from plasma into the injured area20. The
pain, sensitivity to agents with modest analgesic
enzyme responsible for the biosynthesis of the most
activity, and sensitivity to commonly used analgesics
important inflammatory mediators “prostaglandins”
like NSAIDs22. The findings of our investigation
from the natural precursor arachidonic acid is called
showed the ability of P. punctuata leaf ethyl acetate
Cyclooxygenase (COX). There are two types of COX
and ethanol extracts to significantly raise pain
available in human tissues. COX-1 produces basal
threshold as compared to control as shown by the
amounts of physiological prostaglandins that are
reduction in limb licking time and frequency. Similar
needed for several hemostatic functions in the body.
to what was observed with the anti-inflammatory effect
COX-2, on the other hand, is induced in response to
(discussed above), the analgesic effect of the extracts
inflammatory conditions to produce inflammatory
was dose-independent.
Table 4: Antipyretic activity of EtOAC and EtOH (400mg/kg) extracts of P. punctulata leaves and Paracetamol
(150mg/kg) in Yeast-induced pyrexia model in rats
Treatment
Groups
Normal Saline
Paracetamol (150 mg/kg)
Ethyl acetate extract
Ethanol extract
Temperature (C°)
BBT
37.98±0.1
37.78±0.3
38.32±0.4
38.08±0.4
0
38.6±0.1*
38.6±0.4*
38.9±0.*
38.7±0.3*
1
38.7±0.5
37.1±0.4ab
37.7±0.2ab
37.6±0.4ab
2
38.7±0.2
37.2±0.3ab
37.6±0.2ab
37.9±0.4ab
3
38.18±0.4b
37.3±0.2ab
37.7±0.4ab
38.3±0.5b
4
38.2±0.4b
37.9±0.2ab
38.0±0.4b
38.1±0.4b
* = P < 0.05 compared to basal body temperature. a = P < 0.05 compared to control groups at zero time, b = P < 0.05 in compared to zero time of
same group. BBT: Basal body temperature.
properties shown by P. punctuata in the present work
provide the support for the use of P. punctuate leaves
for pain, fever and inflammation, as commonly
practiced in traditional medicine. Although, it is not yet
clear where this medicinal activity comes from, it has
been reported that phenolic compounds inhibit
prostaglandin synthesis and thus elicit an analgesic
effect26. In addition, natural antioxidants (e.g. tannins,
flavonoids) have the potential to bind free radicals
released by leukocytes in response to tissue injury, and
thus may suppress inflammation and pain induced by
these radicals, resulting in decreased pain sensation.
Other potential contributors to the activity arises from
the presence of saponins as previous reports on other
plant species indicated that saponins exert both
analgesic and anti-inflammatory activities. Natural
products are often looked to as a promising source for
bioactive agents with superior safety profile, compared
to synthetic drugs. The present work showed that
ethanol extracts of P. punctulata appeared safe to rats.
Oral administration of ethanol extracts of P. punctulata
leaves for up to 5000 mg/kg resulted in no detectable
toxic signs in body weight, physical activity, behavior,
or mortality rate25. However, people should be cautious
when the leaves are used in oral preparations, until
absolute safety is confirmed in humans.
Pain usually results from tissue damage and is defined
as an unpleasant sensation induced by the release of
endogenous mediators including prostaglandins that are
synthesized by the action of COX on arachidonic acid.
Analgesics usually exert their pharmacological effect
by acting on the central nervous system (CNS) or on
peripheral tissues. Non-steroidal anti-inflammatory
drugs (e.g. diclofenac) and simple analgesics (e.g.
paracetamol) are thought to exhibit their analgesic
activity by blocking biosynthesis of prostaglandins,
either in the CNS or peripherally23,24. Thus, it is
possible that the P. punctulata leaf extracts act by
inhibiting PG synthesis or action, just like simple
analgesics and NSAIDs. Taken into account that
several CNS depressants in high doses have the
potential to produce analgesia by merely suppressing
the brain activity independent of their effect on
prostaglandins and inflammatory mediators25, the test
extracts sedative profile on rats was observed. No
apparent sedative action on rats was noted as shown by
their normal physical activity during the test period.
Therefore, sedation is likely not a contributor to the
analgesic activity of P. punctulata.
In addition to the aforementioned effects, both ethanol
and ethyl acetate extracts of P. punctulata leaves
demonstrated marked anti-pyretic potential as shown
by the inhibiting rise of temperature in the rat yeast
model. The effect was evident in the first two hours but
then faded in the 3rd and 4th hour of the investigation
period, indicating a short antipyretic effect. The
antipyretic effect of the extracts may be ascribed to
inhibiting the synthesis of endogenous pyrogenic
prostaglandins and thus decreasing their levels in
serum and thus in the CNS, especially that these
extracts also exhibited an analgesic and antiinflammatory
potential.
The
pharmacological
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CONCLUSION
In conclusion, the results of this study provide evidence
for the anti-inflammatory, analgesic and antipyretic
activity of P. punctulata leaves claimed in Yemeni folk
medicine. Although the mechanism is not clear, it is
possible that these activities arise from blocking
prostaglandin biosynthesis of prostaglandins by P.
punctulata. These observed pharmacological activities
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may be ascribed to the presence of one or more of the
detected
bioactive
constituents:
alkaloids,
carbohydrates, steroids, poly phenols/tannins, sterols,
saponins, gum and mucilage. What makes this plant
even more promising is its wide margin of safety, as
shown by the rats tolerating up to 5000 mg/kg leaf
ethanol extract. Although this study provides scientific
justification for the ethno-medicinal uses of P.
punctuata, more research needs to be done to ascertain
the safety of the plant in humans and to decipher its
therapeutic molecular mechanisms.
11.
12.
13.
14.
AUTHOR’S CONTRIBUTIONS
All authors participated in designing of experiments,
experimentation, interpretation of data, statistical
analysis, and manuscript writing. Dr Hassan solely
performed the pharmacological experimentation.
15.
ACKNOWLEDGMENT
The authors are thankful to members of pharmacology
and toxicology lab. (Faculty of Pharmacy, University
of Sana’a) for their assistance.
16.
CONFLICT OF INTEREST
No conflict of interest associated with this work.
17.
REFERENCES
18.
1. Midiwo JO, Owuor F, Juma B, Waterman PG. Diterpenes
from the leaf exudate of Psiadia punctulata. Phytochem
1997; 45: 117-120.
https://doi.org/10.1016/S0031-9422(96)00812-6
2. Mabberly Y. The plant book-Cambridge Univ Press, 1987.
3. AlKhulaidi A. Flora of Yemen. Sustainable Natural
resource, Management Project (SNRMP) II. Yemen 2013.
4. Piaz DF, Bader A, Malafronte N, et al. Phytochemistry of
compounds isolated from the leaf-surface extract of
Psiadia punctulata (DC.) Vatke growing in Saudi Arabia.
Phytochem 2018, 155, 191-202.
https://doi.org/10.1016/j.phytochem.2018.08.003
5. Mothana RA, Kriegisch S, Harms M, et al. Assessment of
selected Yemeni medicinal plants for their in vitro
antimicrobial, anticancer, and antioxidant activities. Pharm
Biol 2011; 49: 200-210.
https://doi.org/10.3109/13880209.2010.512295
6. Koch A, Tamez P, Pezzuto J, Soejarto D. Evaluation of
plants used for antimalarial treatment by the Maasai of
Kenya. J Ethnopharmacol 2005; 101: 95-99.
https://doi.org/10.1016/j.jep.2005.03.011
7. Mulwa LS. Phytochemical investigation of Psiadia
punctulata for analgesic agents. Doctoral Dissertation.
University of Nairobi, Kenya, 2012.
http://erepository.uonbi.ac.ke:8080/xmlui/handle/1234567
89/6846
8. Juma BF, Yenesew A, Midiwo JO, Waterman PG.
Flavones and phenylpropenoids in the surface exudate of
Psiadia punctulata. Phytochem 2001; 57: 571-574.
https://doi.org/10.1016/S0031-9422(01)00147-9
9. Moharram BA, Al-Mahbashi HM, Saif-Ali R, Ali AF F.
Phytochemical, anti-inflammatory, antioxidant, cytotoxic
and antibacterial study of Capparis Cartilaginea
Decnefrom yemen. Int J Pharm Pharm Sci 2018; 10: 3834. http://dx.doi.org/10.22159/ijpps.2018v10i6.22905
10. OECD. (Organization for Economic Cooperation and
Development). OECD Guidelines for the Testing of
ISSN: 2456-8058
19.
20.
21.
22.
23.
24.
25.
26.
66
Chemicals—Acute Oral Toxicity-Acute Toxic Class
Method, OECD, Paris, France, 2001.
https://doi.org/10.1787/20745788
Chan PK. Principles and methods for acute toxicity and
eye irritancy. Principles and methods of toxicology 1989,
169-220.
Carvalho K, Vialle E, Moreira G, et al. Functional
outcome of bone marrow stem cells (CD45+/CD34−) after
cell therapy in chronic spinal cord injury in wistar rats.
Transplantation Proceedings 2008; 40: 845-846.
https://doi.org/10.1016/j.transproceed.2008.02.054
Shripad B, Abhijeet A, Inayat P, Nema, N. Analgesic and
anti-inflammatory evaluation of Ficus microcarpa L.
leaves extract. Asian J Pharm Clin Res 2012; 5: 258-261.
Vasundra D, Divya P. Antipyretic activity of ethanol and
aqueous extract of root of Asparagus racemosus in yeast
induced pyrexia. Asian J Pharm Clin Res 2013; 6: 190193.
Subedi NK, Rahman S, Akbar MA. Analgesic and
antipyretic activities of methanol extract and its fraction
from the root of Schoenoplectus grossus. Evidence-Based
Comp Alt Med 2016; 2016.
https://doi.org/10.1155/2016/3820704
Hosseinzadeh H, Younesi HM. Antinociceptive and antiinflammatory effects of Crocus sativus L. stigma and petal
extracts in mice. BMC Pharmacol 2002; 2, 7.
https://doi.org/10.1186/1471-2210-2-7
Igbe I, Ching FP, Eromon A. Anti-inflammatory activity
of aqueous fruit pulp extract of Hunteria umbellata K.
Schum in acute and chronic inflammation. Acta Pol Pharm
2010, 67, 81-85. PMID: 20210083
Fu KY, Light AR, Maixner W. Long-lasting inflammation
and long-term hyperalgesia after subcutaneous formalin
injection into the rat hind paw. The J Pain 2001; 2: 2-11.
https:// doi:10.1054/jpai.2001.9804.
Damas J, Liegeois JF. The inflammatory reaction induced
by formalin in the rat paw. Naunyn- Schmiedeberg's Arch
Pharmacol 1999, 359, 220-227.
https://doi.org/10.1007/PL00005345
Iwalewa E, McGaw L, Naidoo V, Eloff J. Inflammation:
the foundation of diseases and disorders. A review of
phytomedicines of South African origin used to treat pain
and inflammatory conditions. African J Biotech 2007; 6.
https://doi:10.5897/AJB2007.000-2457
Hinz B, Brune K, Pahl A. Cyclooxygenase-2 expression in
lipopolysaccharide-stimulated human monocytes is
modulated by cyclic AMP, prostaglandin E2, and
nonsteroidal anti-inflammatory drugs. Biochem Biophys
Res Comm 2000; 278: 790-796.
https://doi.org/10.1006/bbrc.2000.3885
Prad W, Tonussi C, Rego E, Corrado A. Antinociception
induced by intraperitoneal injection of gentamicin in rats
and mice. Pain1990; 41, 365-371.
https://doi.org/10.1016/0304-3959(90)90013-4
Bensemana D, Gascon A. Relationship between analgesia
and turnover of brain biogenic amines. Canadian J Physiol
Pharmacol 1978; 56, 721-730.
https://doi.org/10.1139/y78-115
Glazer E, Steinbusch H, Verhofstad A, Basbaum A.
Serotonin neurons in nucleus raphe dorsalis and
paragigantocellularis of the cat contain enkephalin. J De
Physiol 1981; 77: 241-245.
Rang H, Rang and Dale's pharmacology. 2012. Edinburgh
etc.: Elsevier/Churchill Livingstone.
Hosseinzadeh H, Sadeghnia HR, Rahimi A. Effect of
safranal on extracellular hippocampal levels of glutamate
and aspartate during kainic acid treatment in anesthetized
rats. Planta Medica 2008; 74: 1441-1445.
https:// DOI: 10.1055/s-2008-1081335
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