Pharmacotherapy and Pharmascience Discovery 2021 1(2) 74-81

http://dx.doi.org/10.13140/RG.2.2.22195.35360

ORIGINAL
CONTRIBUTION

Central and peripheral antinociceptive

activity of D. linearis ethanolic leaf extract

Arif Hossin1 , Mohammed Sahek Ullah Jabed2 , Razwanur Rahman Tushar3 , Kashfia Nawrin3 ,
Mohammad Mustakim Billah3

1PharmacyDiscipline, Khulna University, Khulna, Bangladesh

2Department of Pharmaceutical Sciences, North South University, Dhaka, Bangladesh
3Department of Research, Institute for Pharmaceutical Skill Development and Research, Dhaka, Bangladesh

ABSTRACT
Background: Dicranopteris linearis, a medicinal plant, is considered effective in
relieving pain. Though previously it has been scientifically evaluated, very few of the Keywords
studies aimed at differentiating the central and peripheral analgesia produced by this
plant. The present study was designed to examine the efficacy of its leaf extract in Dicranopteris linearis,
Antinociceptive,
antinociception and qualitatively assess its mode of activity.
Central analgesic,
Methods: Ethanol extract of the leaf at 400, 200 and 100 mg/kg were investigated on Peripheral analgesic
mice and compared with the standard(s). Tail flick, tail pressure, tail immersion and
hot plate methods were employed to observe central acting potential whereas
abdominal constriction and biphasic pain models were executed to understand its
Article Info
peripheral action.
Received: August 26, 2021
Results: DLET 400 mg/kg confirmed moderate efficacy in comparison with the
Revised: October 16, 2021
standard morphine in all centrally acting models and superseded (44.29% in formalin Accepted: October 25, 2021
treatment, late phase) aspirin in peripherally acting models. Lower doses of the extract Published: November 11, 2021
were able to produce mild effects in the experiments. Morphine inhibited the tail flick
response up to 83.45% at 60-minute interval whereas aspirin exhibited similar
efficacy in both writhing (41.06%) and biting (40.35% in late phase) tests. Correspondence

Conclusion: Findings suggested that D. linearis leaf on ethanol extraction yielded in [email protected]
compounds that has potential to suppress nociception. The extract acted more like a
peripheral inhibiting agent. However, further investigation is necessary to prescribe
its safe and optimum use as an analgesic.

Publisher’s Note: Pharmacotherapy and Pharmascience Discovery stays neutral with regard to jurisdictional claims in
published maps and institutional affiliations. © The Author(s). 2021 Open Access. This article is licensed under a Creative
Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in
any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the
Creative Commons licence, and indicate if changes were made.

www.ptherpscid.com 74
Hossin et al. Central and peripheral analgesic activity of D. linearis leaf

INTRODUCTION gm of crude extract was obtained and preserved in air-

tight amber glass container [5].
A significant part of the world population is affected by
pain which exerts a crucial challenge in clinical medicine Drugs and Reagents
[1,2]. As currently available antinociceptive drugs are
Ethanol (RCI Labscan Limited, Thailand), Morphine
associated with adverse effects like sedation, addiction,
(UniMed UniHealth Limited, BD), Acetyl Salicylic Acid
nausea, apnea, constipation etc., and the alternate choices
(Square Pharmaceuticals Limited, BD), Paracetamol
of NSAIDs risks stroke, myocardial infarction,
(Beximco Pharmaceuticals Limited, BD) were obtained
gastrointestinal ulceration and bleeding [1], Current
for the experiments.
studies are focused on finding safer and potent
alternatives. The search includes natural sources like Grouping of Animal
plant materials [3]. Among the many traditionally used
Swiss Albino mice of both sexes, aged 45 days, weighed
plants for analgesic properties, Dicranopteris linearis L., is
24-30g, were selected for the experiments. Mice were
considered a potent one [4]. D. linearis, belonging to
kept in temperature-controlled room at 25±1 °C with 12h
Gleicheniaceae family, is a medicinal plant, known for its
light/dark cycle and fed standard mice pellets and
efficacy over cough, hypersensitivity, respiratory distress,
portable water ad libitum. For each experiment, mice
fever, ulcer, wound, women sterility, intestinal worms
were divided into five groups each containing six mice
and many more ailments [5-7]. Scientifically, the plant has
and designated as follows: Group 1: Control, administered
been confirmed as pharmacologically active against
with vehicle, water; Group 2: Positive Control/Standards,
nociception, pyrexia, inflammation, infection, oxidation,
administered with Morphine (5mg/kg)/Acetyl Salicylic
hepatotoxicity and cytotoxicity whereas other potential
Acid (ASA) (100mg/kg), depending on the experiment;
activities are still under investigation [7-11].
Group 3: Test Sample, D. linearis ethanolic leaf extract
As a traditionally used natural pain reliever, D. linearis (DLET) 100 mg/kg; Group 4: Test Sample, DLET 200
was previously investigated at different solvent mg/kg; Group 5: Test Sample, DLET 300 mg/kg.
extraction (aqueous, methanol and chloroform) [8,12]. As
Acute Toxicity Test
selection of solvent extracts polar to non-polar plant
material, the present study was attempted to fractionate To investigate the immediate and short-term toxicity,
the polar compounds of the plant leaf with ethanol and healthy mice (n=5) were orally administered with high
evaluate its analgesic activity in animal model [13]. No doses (100, 250, 500, 1000 mg/kg) of the plant extract
such study was reported before. Moreover, very few and observed for the next 3 days for unusual behavior or
studies were found which explored both peripheral and any mortality [14].
central analgesia simultaneously. Thus, the study was
Central Antinociceptive Tests
also aimed to validate its appropriate use by comparing
the central and peripheral analgesic action of the plant. Tail pressure method
Alongside the scientific evaluation of the efficacy, the
In this method, mechanical pain was induced at the base
study was also focused to draw optimum dose line for
of mice tail through applying metal artery clip having its
traditional use of the plant for antinociception.
jaw covered with silicon to avoid tissue damage. The
METHODS method was applied before and after the drug
administration [15]. A cutoff time of 10s was considered
Collection and preparation of the extract and mice which did not struggle to get rid of the clip were
Fresh plant leaf (approximately 7 kg) was collected from not selected for further experiment. The time at which
Mymensingh district (24°45’14” N 90°24’11” E) of mice attempted to extricate the clip was recorded. The
Bangladesh in June. 2014. A sample specimen was process was repeated at 30, 60, 90 and 120 minutes after
submitted to Bangladesh National Herbarium and the drug or test sample administration. Morphine
preserved with an accession number DACB 42009. After (5mg/kg) served as positive control. From the
a thorough wash, the leaves were sundried before comparison of pre- and post-treatment, percentage
crushed into powder. Approximately, 600g powder was inhibition of pain was calculated. Increase in pain
obtained and soaked in 3L of ethanol (96%) and left for threshold in comparison with the control group was
72h with occasional shaking. At the end, the mixture was considered indication for antinociceptive activity.
sieved with paper filter and concentrated using a Rotary 𝑇𝑠𝑎𝑚𝑝𝑙𝑒 −𝑇𝐶𝑜𝑛𝑡𝑟𝑜𝑙
Evaporator (Biobase RE-2010, China). Approximately 3 Percentage inhibition of pain (%) = × 100
𝑇𝐶𝑜𝑛𝑡𝑟𝑜𝑙

www.ptherpscid.com 75
Pharmacotherapy and Pharmascience Discovery 2021 1(2) 74-81
http://dx.doi.org/10.13140/RG.2.2.22195.35360

Where T = the time at which mice attempted to extricate Percentage inhibition of pain (%) =
𝑇𝑠𝑎𝑚𝑝𝑙𝑒 −𝑇𝐶𝑜𝑛𝑡𝑟𝑜𝑙
× 100
𝑇𝐶𝑜𝑛𝑡𝑟𝑜𝑙
its tail.
Where, T=time.
Tail flick response method
Peripheral Antinociceptive Tests
In this experiment, radiant heat was applied to mice tail
using an analgesiometer (Orchid Scientific TFA01, India) Acetic Acid-Induced Abdominal Constriction Test
to record the tail flicking latency before and after the drug
or test sample administration [16,17]. The mice were kept The acetic acid induced abdominal constriction test was
individually in suitable restrainer keeping the tail free. 5A conducted to evaluate the peripheral antinociceptive
current was passed through a naked nichrome wire activity of the plant extract [19]. 0.6% acetic acid
where the tail was placed at a distance of 1.5 cm and (0.1ml/10g) was peritoneally injected to mice before they
applied within 2 cm of the tail. The time between the were observed for specific pattern of abdominal
onset of heat application and flicking of the tail was noted constriction, also known as writhing, for 25 minutes
as reaction time. To avoid tissue injury, a cut-of time of keeping the first 5 minutes excluded from the calculation.
10s was considered. Paracetamol (10mg/kg) served as Standard ASA (100mg/kg) and the test samples were
standard. The withdrawal time for the groups were administered 1h prior commencing the experiment.
compared with the negative control group where Decrease in number of writhing was considered indicator
prolongation of the flicking response was considered as of antinociceptive activity. Percentage inhibition was
indication of antinociceptive activity. Percentage calculated using the following formula:
inhibition of pain was deduced from below formula: 𝑊𝐶𝑜𝑛𝑡𝑟𝑜𝑙 −𝑊𝑆𝑎𝑚𝑝𝑙𝑒
Percentage inhibition of pain (%) = 𝑊𝐶𝑜𝑛𝑡𝑟𝑜𝑙
× 100
𝑇𝑠𝑎𝑚𝑝𝑙𝑒 −𝑇𝐶𝑜𝑛𝑡𝑟𝑜𝑙
Percentage inhibition of pain (%) = × 100
𝑇𝐶𝑜𝑛𝑡𝑟𝑜𝑙 Where, W= number of writhing activities.
Where, T=time. Formalin-Induced Paw Licking Test.
Tail Immersion Method The formalin induced paw licking and biting test had been
described as an appropriate model for assessing both
Mice tail at 2cm at the tip was dipped into warm water
central and peripheral analgesic properties of medicinal
bath (55.0 ± 0.5 C) and the reaction time to withdraw the
agents [18,19]. In this test, a centrally acting analgesic
tail was recorded immediately before and in every 30
(ASA 100mg/kg) and a central and peripheral acting
minutes interval after drug administration for 2 hours
analgesic (Morphine 5mg/kg) was applied as standards.
[18]. Longer sustained duration for tail withdrawal was
Sixty minutes after the drug and test sample
considered an indication by the sample as pain reliever.
administration, 20 µl of 5% v/v formalin was injected in
Percentage inhibition was measured using following
subplantar surface of the left hind paw. The licking and
equation:
biting responses were measured in seconds at two phases
Percentage inhibition of pain (%) =
𝑇𝑠𝑎𝑚𝑝𝑙𝑒 −𝑇𝐶𝑜𝑛𝑡𝑟𝑜𝑙
× 100 – early phase (0-5 min) and the late phase (16-30 min).
𝑇𝐶𝑜𝑛𝑡𝑟𝑜𝑙
reduction in biting and licking activity compared to the
Where, T=time. control group was considered the indication of analgesia.
Percentage of pain inhibition was calculated as:
Hot Plate Test
𝐿𝑇𝐶𝑜𝑛𝑡𝑟𝑜𝑙 −𝐿𝑇𝑆𝑎𝑚𝑝𝑙𝑒
Percentage inhibition of pain (%) = × 100
Thermal induced nociceptive stimulus was applied in this 𝐿𝑇𝐶𝑜𝑛𝑡𝑟𝑜𝑙

test using a hot plate, where mice were placed and Where, LT= total duration of paw licking activity.
observed for their escape-oriented behavior before and
after the drug administration [19]. At first, a Ugo Basile Statistical Analysis
7280 hotplate, Italy was heated to 50 ± 0.2 °C and mice
Maximum possible analgesia (MPA) was calculated in
were placed and selected for the main test based on the
percentage and presented from data as mean ± standard
cut-off latency of 5-7s. after selection, mice were
error of the reaction time in all tests except for the
challenged pre- and post- oral drug treatment at 30-, 60-,
abdominal constriction test where the data was based on
90- and 120-minutes interval with the hotplate and
the number of writhing. All groups were compared to the
response time was recorded. Percentage inhibition was
negative control group and in this regard, one way
calculated using the general formula:
analysis of variance test (ANOVA) was performed

www.ptherpscid.com 76
Hossin et al. Central and peripheral analgesic activity of D. linearis leaf

followed by Dunnett’s T test via SPSS for windows Tail pressure Test
software (version 24). Data were considered statistically
In tail pressure test, maximum pendency was observed by
significant when the confidence interval was found at
morphine at 60 min (79.41%) followed by smooth fall in
95% (p<0.05), 99% (p<0.01) or 99.99% (p<0.001),
action (Figure 1a). On the other hand, DLET 400
denoted with asterisk (*) sign.
demonstrated a linear increase in efficacy till 120 min.
RESULTS DLET 200 mimicked its higher dose though DLET 100 was
associated with a sharp fall after its peak at 60 min
Acute Toxicity Test (29.25%) as exhibited by morphine.
In the period of treatment and its following observation,
Tail flick response Test
no abnormal behavior or symptoms or death was
recorded. However, indigestion was reported at higher Findings of the experiments were graphically presented
doses like 1000 mg/kg. As a consequence, lower doses to compare the test samples with the standard drug. In tail
were adopted for the main experiments. flick test, morphine at 5 mg/kg showed highest peak in

a TAIL PRESSURE TEST b TAIL FLICK TEST

MOR 5 DLET 100 DLET 200 DLET 400 MOR 5 DLET 100 DLET 200 DLET 400

100 100 ***

MAXIMUM POSSIBLE ANALGESIA (%)
MAXIMUM POSSIBLE ANALGESIA (%)

90 *** 90
** **
80 80 *
**
70 *
70
60 60 **
50 50
40 40
30 30
20 20
10 10
0 0
0 MIN 30 MIN 60 MIN 90 MIN 120 MIN 0 MIN 30 MIN 60 MIN 90 MIN 120 MIN

c TAIL IMMERSION TEST d HOT PLATE TEST

MOR 5 DLET 100 DLET 200 DLET 400 MOR 5 DLET 100 DLET 200 DLET 400

100 100
MAXIMUM POSSIBLE ANALGESIA (%)
MAXIMUM POSSIBLE ANALGESIA (%)

*** ***
90 90 **
* ***
80 ** 80 *
70 70
*
60 60
50 50
40 40
30 30
20 20
10 10
0 0
0 MIN 30 MIN 60 MIN 90 MIN 120 MIN 0 MIN 30 MIN 60 MIN 90 MIN 120 MIN

Figure 1 (a-d): Maximum possible analgesia (MPA) (%) representing the effect of the ethanol extract of the leaf of D. linearis compared
to morphine sulfate (positive control) administered into mice, evaluated by centrally-acting models of (a) tail- pressure method (b)
tail-flick method (c) tail immersion method and (d) hot plate method. MOR = Morphine Sulphate, DLET = D. linearis leaf ethanol
extract. Data presented as mean ± standard error (n=6). and analyzed by one-way ANOVA followed by Dunnett t test where *, **, ***
denoted p < 0.05, p < 0.01 and p < 0.001 respectively and statistically significant. All groups were compared to control.

www.ptherpscid.com 77
Pharmacotherapy and Pharmascience Discovery 2021 1(2) 74-81
http://dx.doi.org/10.13140/RG.2.2.22195.35360

efficacy at 60 min (83.45%) followed by a slight fall in action after its first rise at 30 min (44.21%). Both DLET
next two intervals (Figure 1b). On the contrary, DLET 400 200 and 100 was observed with increased efficacy till 60
mg/kg exhibited a gradual increase in analgesia till 90 min though could not produce significant differnces in
min (72.95%). DLET at lower doses showed mild action action in late phases (90 min and 120 min).
at initial intervals though shifted to moderate action over
Hot Plate Test
time.
In hot plate method, morphine showed a linear sharp
Tail Immersion Test
increase in analgesia from 30 min till 120 min (68.56% to
Figure 1c depicted that, unlike tail flick and tail pressure 86.24%) (Figure 1d). On the contrary, other groups
methods, in tail immersion test, morphine was found to showed a mild increase in efficay after the first interval
increase its analgesic action till 90 min (77.68%) whearas and ended in downward action in last two intervals (90
DLET 400 was observed with no significant increase in min and 120 min).

a ACETIC ACID INDUCED WRITHING TEST b FORMALIN INDUCED PAW LICK TEST
80 ASA 100 DLET 100 DLET 200 DLET 400 MOR 5
***
MAXIMUM POSSIBLE ANALGESIA (%)

MAXIMUM POSSIBLE ANALGESIA (%)

70 90
60 80 ***

50 ** 70

* 60 **
40
50 * **
30 *
40
20 30
10 20
10
0
ASA 100 DLET DLET DLET MOR 5 0
100 200 400 EARLY PHASE LATE PHASE

Figure 2 (a-b): Maximum possible analgesia (MPA) (%) representing the effect of the ethanol extract of the leaf of D. linearis compared
to morphine sulfate and acetyl salicylic acid (positive controls) administered into mice, evaluated by peripherally-acting models of (a)
acetic acid induced writhing test and (b) formalin induced biphasic pain test. MOR = Morphine Sulphate, ASA = Acetyl Salicylic Acid,
DLET = D. linearis leaf ethanol extract. Data presented as mean ± standard error (n=6). and analyzed by one-way ANOVA followed
by Dunnett t test where *, **, *** denoted p < 0.05, p < 0.01 and p < 0.001 respectively and statistically significant. All groups were
compared to control.

Acetic Acid-Induced Abdominal Constriction Test inhibited the pain up to 27.66% and 26.09% respectively
in early phase whereas in late phase, their action climbed
In acetic acid induced abdominal constriction test, the
up to 68.52% and 44.29% respectively. DLET 200 also
standard ASA 100 was able to reduce the pain by 41.06%
produced a significant reduction of pain in late phase
(Figure 2a) whereas morphine suppressed the pain
(33.74%).
response to a great extent (65.23%). DLET 400 closely
met the standard ASA by its action (32.75%). DLET 200 DISCUSSION
acted as a moderate analgesic (21.31%) whereas DLET
Antinociceptive drugs act on central or peripheral
100 produced mild effect (16.49%).
nervous system in order to alleviate or relieve pain
Formalin-Induced Paw Licking Test however, without significant alteration of consciousness
[20]. While central analgesics raise the threshold for pain
In formalin induced biphasic pain test, all groups and alter physiological response towards it, peripheral
generated mild action in early phases compared to that of analgesics inhibit the impulse generation at
their late phases except for morphine which significantly chemoreceptor sites of pain [21]. In this study, pain-state
inhibited the pain in both phases (41.86% and 68.48% models were employed using thermal and pressure
respectively) (Figure 2b). ASA 100 and DLET 400 stimuli which illustrated the central analgesic responses

www.ptherpscid.com 78
Hossin et al. Central and peripheral analgesic activity of D. linearis leaf

of drugs and test samples focusing above the spinal cord of morphine in abdominal constriction as well as in the
[22]. Tail flick, tail immersion and tail clip models early phase licking activities by the mice.
mediated a spinal reflex to a nociceptive stimulus
The findings indicated that ethanol extract of D. linearis
whereas hot plate method involved supraspinally
leaf acted more as a peripheral acting antinociceptive
organized brain functions of mice [23]. Except the
agent rather than its central action though data supports
pressure model, the other methods were acute thermic
its moderate efficacy on central nervous system. The leaf
and phasic pain model which were subjected to selective
contains high amount of phenol, flavonoids (as flavonol 3-
attenuation of centrally-acting opioid-like analgesic
Oglycosides), triterpenes, saponins and steroids [5].
compounds [18]. Regarding the peripheral analgesic
Moreover, previous studies reported possible
models, acetic acid induced abdominal constriction was
mechanisms for central and peripheral analgesic activity
caused by peritoneal tissue damage and induced
involving modulation of opioid receptors and TRPV1,
inflammation by peritoneal macrophages and mast cells
bradykininergic and glutamatergic system, PKC activity
released by TNF-α, IL-1β, IL-8, bradykinin, substance P,
and l-arginine/NO-dependent, cGMP-independent
serotonin and histamine like mediators [24,25]. Analgesic
pathway [4]. Alongside, numerous volatile and non-
activity expressed in this model were due to involvement
volatile bioactive compounds was found in literature
of α2 and β1 adrenergic receptors [26]. On the contrary,
suggesting attenuation of nociception in mice [4].
formalin induced biphasic pain model was established
through two phase nociceptive responses, firstly, by
CONCLUSION
sensitizing sensory C-fibers and at prolonged phase by
developing injury-induced spinal sensitization which In both peripheral and central acting models of
eventually sensitized dorsal horn neuron for antinociceptive studies, D. linearis leaf demonstrated
inflammation associated pain [18,27]. In general, efficacy though the degree of effectiveness were
centrally-acting drugs, like morphine, suppresses both moderate. At this stage of study, responsible biological
phases of pain, while peripherally-acting drugs like compounds were not investigated. Therefore, further
aspirin only suppress the late phases [28]. investigations might be directed to determine optimum
doses, best extraction solvent and phytochemical
Morphine sulphate and Acetyl Salicylic Acid were used as
screening and fractionation.
the standards as central and peripheral acting analgesics
respectively. Morphine is a centrally-acting opioid-like Abbreviations
analgesic compound whereas Aspirin is peripheral
NSAIDs [4,24,29]. Being non-selective, aspirin NSAIDs: Non-Steroidal Anti-Inflammatory Drugs; ASA:
irreversibly blocks cyclooxygenase isozymes- COX-1 and Acetyl Salicylic Acid; DLET: D. linearis leaf ethanol extract;
COX-2 that generates prostaglandin, a proinflammatory TNF: Tumor Necrosis Factor; IL: Interleukins; TRPV1: The
substance [30]. On the other hand, morphine binds to transient receptor potential cation channel subfamily V
opioid receptors, inhibits transmission of pain signals, member 1; cGMP: Cyclic Guanosine Monophosphate.
signals nociception-modulating neurons in the spinal Acknowledgments
cord, and also blocks primary afferent pain receptors to
the dorsal horn sensory projection cells [31]. In all the The present study was supported and carried out in the
experiments, morphine exhibited most significant Pharmacology lab of Institute for Pharmaceutical Skill
antinociceptive response among all the groups at all Development and Research, Bangladesh. Authors are
intervals in comparison to control. In the central analgesic grateful to the institution for providing such opportunity
study through tail flick, tail pressure, tail immersion and to contribute to health science.
hot plate methods, D. linearis leaf extract demonstrated
Authors’ Contributions
significant pain reduction through prolonged reaction
time by its high dose (400 mg/kg) however, failed to This work was carried out in collaboration between all
exhibit strong analgesic action by lower doses (200 & 100 authors. Authors KN and MMB designed, coordinated and
mg/kg) as compared to morphine (5mg/kg). While in supervised the project. AH, MSUJ and RRT performed the
peripheral acting models, DLET 400 nearly produced experiments and prepared the graphical presentations.
similar level of action as aspirin (100 mg/kg) though the KN prepared the manuscript. MMB participated in the
other two smaller doses could not produce directly interpretation of data to reach a scientific discussion. All
comparable efficacy. None could reach the effectiveness authors read and approved the final manuscript.

www.ptherpscid.com 79
Pharmacotherapy and Pharmascience Discovery 2021 1(2) 74-81
http://dx.doi.org/10.13140/RG.2.2.22195.35360

Funding Dicranopteris linearis L. leaf. Clinical Phytoscience.

2021;7(1):25. doi:10.1186/s40816-021-00262-8
This research did not receive any specific grant from
funding agencies in the public, commercial, or not-for- 6. Sarker SK, Hossain ABME. Pteridophytes of greater
Mymensingh district of Bangladesh used as
profit sectors. The study was carried out with individual
vegetables and medicines. Bangladesh Journal of
funding of all authors. Plant Taxonomy. 2009;16(1):47-56. doi:10.3329/
bjpt.v16i1.2746
Availability of Data and Materials
7. Hussaini J, Othman NA, Abdulla MA, Majid NA, Faroq
The datasets used and/or analyzed during the current HM, Ismail S. Gastroprotective effects of
study are available from the corresponding author on Dicranopteris linearis leaf extract against ethanol-
reasonable request. induced gastric mucosal injury in rats. SRE.
2012;7(18):1761-1767. doi:10.5897/SRE11.775
Ethics Approval and Consent to Participate
8. Zakaria ZA, Abdul Ghani ZDF, Raden Mohd Nor RNS,
All experiments associated with animal handling were Gopalan HK, Sulaiman MR, Abdullah FC.
performed in accordance with the Guide for the Care and Antinociceptive and anti-inflammatory activities of
Dicranopteris linearis leaves chloroform extract in
Use of Laboratory Animals, 8th ed.; The National
experimental animals. Yakugaku Zasshi.
Academies Collection adopted by the institutional 2006;126(11):1197-1203. doi:10.1248/yakushi.126.
guideline for animal handling (Ref. no. IPSDRLAB/AHCP/ 1197
01/18). The experimental design was authorized by the
9. Ismail NA, Shamsahal-Din NS, Mamat SS, et al. Effect
Institutional Ethical Committee Clearance (Ref. No. of aqueous extract of Dicranopteris linearis leaves
IPSDRLAB/IECC/11/20) from the Institute for against paracetamol and carbon tetrachloride-
Pharmaceutical Skill Development and Research, induced liver toxicity in rats. Pak J Pharm Sci.
Bangladesh. 2014;27(4):831-835.

Consent for Publication 10. Lai HY, Lim YY, Tan SP. Antioxidative, tyrosinase
inhibiting and antibacterial activities of leaf extracts
Not applicable. from medicinal ferns. Biosci Biotechnol Biochem.
2009;73(6):1362-1366. doi:10.1271/bbb.90018
Competing Interests
11. Zakaria ZA, Mohamed AM, Jamil NM, et al. In vitro
All authors agreed on the article before submission and cytotoxic and antioxidant properties of the aqueous,
chloroform and methanol extracts of Dicranopteris
had no conflict of interests.
linearis leaves. African Journal of Biotechnology.
REFERENCES 2011;10(2):273-282. doi:10.4314/ajb.v10i2
12. Zakaria ZA, Ghani ZDFA, Nor RNSRM, et al.
1. Del Vecchio G, Spahn V, Stein C. Novel Opioid Antinociceptive, anti-inflammatory, and antipyretic
Analgesics and Side Effects. ACS Chem Neurosci. properties of an aqueous extract of Dicranopteris
2017;8(8):1638-1640. doi:10.1021/acschemneuro.7 linearis leaves in experimental animal models. J Nat
b00195 Med. 2008;62(2):179-187. doi:10.1007/s11418-007-
2. Manchikanti L, Kaye AM, Kaye AD. Current State of 0224-x
Opioid Therapy and Abuse. Curr Pain Headache Rep. 13. Billah MM, Nawrin K, Ahmed KT, Jabed MSU, Islam
2016;20(5):34. doi:10.1007/s11916-016-0564-x MN, Uddin MM. GABA mediated response of aqueous,
3. Dias DA, Urban S, Roessner U. A Historical Overview ethanol and ethyl acetate extracts of Dicranopteris
of Natural Products in Drug Discovery. Metabolites. linearis leaf in Swiss Albino mice. J Herbmed
2012;2(2):303-336. doi:10.3390/metabo2020303 Pharmacol. 2015;5(1):1-6.

4. Zakaria ZA, Roosli RAJ, Marmaya NH, Omar MH, Basir 14. Rayhan MA, Vabna NJ, Ahmed F, Hossin A, Nawrin K,
R, Somchit MN. Methanol Extract of Dicranopteris Billah MM. Jujube (Ziziphus jujube) honey treats
linearis Leaves Attenuate Pain via the Modulation of stress induced anxiety behavior in mice.
Opioid/NO-Mediated Pathway. Biomolecules. 2020; Pharmacotherapy and Pharmascience Discovery.
10(2):E280. doi:10.3390/biom10020280 2021;1(1):1-9. doi:10.13140/RG.2.2.30933.55521
5. Billah MM, Chowdhury AS, Nawrin K, Mostaq S, 15. Singh S, Majumdar DK. Analgesic Activity of Ocimum
Rayhan MdA, Tushar RR. Serotonergic and sanctum and its Possible Mechanism of Action.
noradrenergic response of ethanol extract; International Journal of Pharmacognosy. 1995;
opioidergic response of ethyl acetate extract of 33(3):188-192. doi:10.3109/13880209509065361

www.ptherpscid.com 80
Hossin et al. Central and peripheral analgesic activity of D. linearis leaf

16. Ramesh R. Analgesic Effects of the Aqueous Extracts 26. Bhaskar M, Jagtap AG. Exploring the possible
of Plant Ipomea pes-tigridis Studied in Albino Mice. mechanisms of action behind the antinociceptive
Global J Pharmacol. 2010;4(1):31–5. activity of Bacopa monniera. Int J Ayurveda Res.
2011;2(1):2-7. doi:10.4103/0974-7788.83173
17. Tarkang PA, Okalebo FA, Siminyu JD, et al.
Pharmacological evidence for the folk use of Nefang: 27. Ashok P, Prasanna GS, Mathuram V. Analgesic and
antipyretic, anti-inflammatory and antinociceptive anti-inflammatory activity of the chloroform extract
activities of its constituent plants. BMC Complement of Trichilia connatoides (W&A) Bentilien. Indian J
Altern Med. 2015;15:174. doi:10.1186/s12906-015- Pharm Sci. 2006;68:231–233.
0703-7
28. da Rocha CQ, Vilela FC, Cavalcante GP, et al. Anti-
18. Yemitan OK, Adeyemi OO. Mechanistic assessment of inflammatory and antinociceptive effects of
the analgesic, anti-inflammatory and antipyretic Arrabidaea brachypoda (DC.) Bureau roots. J
actions of Dalbergia saxatilis in animal models. Pharm Ethnopharmacol. 2011;133(2):396-401. doi:10.1016
Biol. 2017;55(1):898-905. doi:10.1080/13880209. /j.jep.2010.10.009
2017.1283706
29. Satyanarayana PSV, Jain NK, Singh A, Kulkarni SK.
19. Sani MHMohd, Zakaria ZA, Balan T, Teh LK, Salleh MZ. Isobolographic analysis of interaction between
Antinociceptive Activity of Methanol Extract of cyclooxygenase inhibitors and tramadol in acetic
Muntingia calabura Leaves and the Mechanisms of acid-induced writhing in mice. Prog
Action Involved. Evid Based Complement Alternat Neuropsychopharmacol Biol Psychiatry. 2004;28(4):
Med. 2012;2012:890361. doi:10.1155/2012/890361 641-649. doi:10.1016/j.pnpbp.2004.01.015
20. Fan S-H, Ali NA, Basri DF. Evaluation of Analgesic 30. Sharma S, Sharma SC. An update on eicosanoids and
Activity of the Methanol Extract from the Galls of inhibitors of cyclooxygenase enzyme systems. Indian
Quercus infectoria (Olivier) in Rats. Evid Based J Exp Biol. 1997;35(10):1025-1031.
Complement Alternat Med. 2014;2014:976764.
31. Pacifici GM: Metabolism and pharmacokinetics of
doi:10.1155/2014/976764
morphine in neonates: A review. Clinics (Sao Paulo).
21. Shreedhara CS, Vaidya VP, Vagdevi HM, Latha KP, 2016 Aug;71(8):474-80. doi: 10.6061/clinics/2016
Muralikrishna KS, Krupanidhi AM. Screening of (08)11.
Bauhinia purpurea Linn. for analgesic and anti-
inflammatory activities. Indian J Pharmacol.
2009;41(2):75-79. doi:10.4103/0253-7613.51345
22. Wigdor S, Wilcox GL. Central and systemic morphine-
induced antinociception in mice: contribution of Submit your Manuscript to Pharmacotherapy
descending serotonergic and noradrenergic and Pharmascience Discovery journal and
pathways. J Pharmacol Exp Ther. 1987;242(1):90-95. get benefitted from:

23. Chapman CR, Casey KL, Dubner R, Foley KM, Gracely ✓ Rolling publication schedule
RH, Reading AE. Pain measurement: an overview. ✓ Rigorous and constructive peer review
Pain. 1985;22(1):1-31. doi:10.1016/0304-3959(85) ✓ Quick peer-review period
90145-9 ✓ Open Access: freely available online
✓ Minimal waiting period from acceptance
24. Ribeiro RA, Vale ML, Thomazzi SM, et al. Involvement
to publication
of resident macrophages and mast cells in the
writhing nociceptive response induced by zymosan
and acetic acid in mice. Eur J Pharmacol. Submit your manuscript at ptherpscid.com
2000;387(1):111-118. doi:10.1016/s0014-2999(99)
00790-6
25. Wang QS, Yang L, Cui WY, Chen L, Jiang YH. Anti-
Inflammatory and Anti-Nociceptive Activities of
Methanol Extract from Aerial Part of Phlomis
younghusbandii Mukerjee. PLOS ONE. 2014;9(3):
e89149. doi:10.1371/journal.pone.0089149

www.ptherpscid.com 81