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Department of Biological & Biomedical Sciences
Medical College, Pakistan
February 2010
The antinociceptive activity of Polygonatum
verticillatum rhizomes in pain models
Haroon Khan
Muhammad Saeed
Anwar Gilani
Aga Khan University
Murad Ali Khan
Ahsana Dar
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Recommended Citation
Khan, H., Saeed, M., Gilani, A., Khan, M., Dar, A., Khan, I. (2010). The antinociceptive activity of Polygonatum verticillatum
rhizomes in pain models. Journal of Ethnopharmacology, 127(2), 521-527.
Available at: http://ecommons.aku.edu/pakistan_fhs_mc_bbs/141
Authors
Haroon Khan, Muhammad Saeed, Anwar Gilani, Murad Ali Khan, Ahsana Dar, and Inamullah Khan
This article is available at eCommons@AKU: http://ecommons.aku.edu/pakistan_fhs_mc_bbs/141
Journal of Ethnopharmacology 127 (2010) 521–527
Contents lists available at ScienceDirect
Journal of Ethnopharmacology
journal homepage: www.elsevier.com/locate/jethpharm
The antinociceptive activity of Polygonatum verticillatum rhizomes in
pain models
Haroon Khan a , Muhammad Saeed a,∗ , Anwal-Ul-Hassan Gilani b , Murad Ali Khan c ,
Ahsana Dar d , Inamullah Khan a
a
Department of Pharmacy, University of Peshawar, Peshawar 25120, Pakistan
Department of Biological and Biomedical Sciences, Aga Khan University, Stadium Road, P.O. Box 3500, Karachi 74800, Pakistan
Department of Chemistry, Kohat University of Science and Technology, Kohat, Pakistan
d
International Centre for Chemical Sciences, HEJ Research Institute of Chemistry, University of Karachi, Karachi 75270, Pakistan
b
c
a r t i c l e
i n f o
Article history:
Received 14 May 2009
Received in revised form
10 September 2009
Accepted 5 October 2009
Available online 22 October 2009
Keywords:
Polygonatum verticillatum
Liliaceae
Antinociceptive
Diuretic
Saponins
Alkaloids
a b s t r a c t
Aim of the study: The current study was designed to establish the pharmacological rationale for the
traditional use of the rhizomes of Polygonatum verticillatum in the treatment of painful conditions and as
a plant diuretic.
Materials and methods: The crude methanolic extract of the rhizomes of Polygonatum verticillatum (PR)
was tested in various established pain models in rodents at 50, 100 and 200 mg/kg i.p. while the diuretic
activity was assessed at 300 and 600 mg/kg p.o. in rats.
Results: PR demonstrated significant reduction (14–72%) in the number of writhes induced by acetic acid
in a dose-dependent manner. When nociceptive threshold was measured in the formalin test, PR strongly
attenuated the formalin-induced flinching behaviour in both phases (6–30% in first phase while 12–72%
in second phase). Central involvement in the analgesic profile of PR was confirmed by the hot plate test, in
which PR elicited a significant (P < 0.01) analgesic activity by increasing latency time. However, an opioid
receptor antagonist, naloxone (2 mg/kg s.c.) strongly antagonized the antinociceptive activity of PR. As
a plant diuretic, PR showed mild but statistically insignificant diuretic activity at 300 mg/kg. The crude
extract and solvent fractions of the plant contained reasonable quantity of total saponin and alkaloid
contents.
Conclusions: The mechanisms underlying the analgesic action of PR shows that the opioid dependant
central mediation has synergistic effect by enforcing the peripheral analgesic effects. Interestingly, our
findings not only substantiated the folk use of the plant as an analgesic but also reported for the first time
in the whole genus.
© 2009 Elsevier Ireland Ltd. All rights reserved.
1. Introduction
Polygonatum (King Solomon’s-seal, Solomon’s Seal) is a genus of
about 57 species belongs to family Liliaceae or Convallariaceae. It
is widely distributed in East Asia, mainly China and Japan where
40 species of Polygonatum are found (Tamura, 1993; Monika et al.,
2006). Solomon’s seal has been used for thousands of years in herbal
medicine. The rhizomes are antiperiodic, antitussive, cardiotonic,
demulcent, diuretic, energizer, hypoglycemic, sedative, tonic and
are used in the treatment of dry coughs and pulmonary problems,
including tuberculosis (Jiang, 1977, 1986; Flora of China, 1980). The
antibacterial and antifungal activity of the Polygonatum has been
reported (Alluri et al., 2006). Polygonatum reduced blood sugar level
∗ Corresponding author. Tel.: +92 919216750.
E-mail address: [email protected] (M. Saeed).
0378-8741/$ – see front matter © 2009 Elsevier Ireland Ltd. All rights reserved.
doi:10.1016/j.jep.2009.10.003
by different mechanisms including ␣-glucosidase inhibition and
increased insulin sensitivity (Li et al., 2004; Hong et al., 2008). The
cytotoxic activity with human MCF-7 breast cancer cells of various
steroidal saponins isolated from the Rhizomes of Polygonatum is
also on the record (Mi-Jeong et al., 2006).
Polygonatum verticillatum [L.] All. (Nooreallam) is a perennial
rhizomatous herb. The rhizomes are usually shortly branched and
0.7–1.5 cm thick. Stem is usually erect and the leaves are four to
eight in a whorl; the flowers are hermaphrodite. The syrup of fresh
rhizome is used in the treatment of pain, pyrexia, burning sensation and for phthisis (Amrit, 2006). It is also used in combination
with other herbs to promote urine discharge (diuretic) and removes
painful urine (Ballabh et al., 2008). Other ethnobotanical uses of
the plant include as emollient, aphrodisiac, vitiated condition of
pitta and vata, appetizer and tonic, galactagogue (increases milk
release), weakness (Ghayur, 2004). It is also used as a substitute
of Polygonatum cirrhifolium (Parveen et al., 2004). Keeping in view
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H. Khan et al. / Journal of Ethnopharmacology 127 (2010) 521–527
the ethnobotanical use of the plant in the treatment of painful
conditions, the current study was designed to provide scientific
background to this claim.
2. Materials and methods
2.1. Plant material
Polygonatum verticillatum All was collected from District Swat
(Gabral and Miandam valleys), N.W.F.P, Pakistan, in July–August
2007. The botanical identity of the plant material was done by
the Taxonomy Department of PCSIR Laboratories Peshawar and
a specimen with catalogue No.: 9970 (PES) was submitted to the
herbarium of PCSIR Laboratories Peshawar.
2.2. Plant extraction and fractionation
The air dried rhizomes of the Polygonatum verticillatum (8 kg)
was ground to fine powder and extracted by maceration with
methanol at room temperature for 14 days with occasional shaking
(Khan et al., 2007, 2009). The methanol soluble residue was filtered
off and concentrated under vacuum at low temperature (40 ◦ C)
using rotary evaporator, yielded a dark greenish semisolid material (2.2 kg, 27.50%, w/w). The crude methanol extract (1.6 kg) was
dissolved in distilled water and sequentially fractionated with hexane, chloroform, ethyl acetate and butanol, yielding hexane (258 g,
16.13%, w/w), chloroform (219 g, 13.69%, w/w), ethyl acetate (226 g,
14.13% w/w), butanol (265 g, 16.56%, w/w), and aqueous (501 g,
31.31%, w/w). These fractions were then screened for various pharmacological and phytochemical analysis.
2.3. Animals
Swiss albino mice (20–25 g) and Wistar rats (210–270 g) of
either sex were used in various experimental models. Animals were
housed 10 per cage and were fed laboratory diet ad libitum and
allowed free access to drinking water under standard environmental condition of temperature (25 ◦ C), relative humidity 60 ± 10% and
light/dark cycles (12/12 h). Experiments were performed according
to ethical principles established in 1979 for laboratory animals at
the service of mankind Lyons, France.
2.4. Drugs and reagents
The chemicals used in this study include: aspirin (Reckitt and
Colman, Pakistan), acetic acid, formalin, morphine sulphate, naloxone hydrochloride, sodium chloride hydrochlorothiazide (Sigma
chemicals company, St. Louis, USA) Sterile normal saline was used
as control in all studies and the crude methanol extract (PR) used
in various studies were prepared in normal saline.
1959; Adzu et al., 2001). Briefly, the prescreened animals were
divided into five groups (n = 6). The writhes were induced by
intraperitoneal injection of 1.0% acetic acid (v/v, 0.1 ml/10 g body
weight). Group I was used as control, received normal saline
(10 ml/kg, i.p.); groups II, III and IV were treated with PR (50,
100 and 200 mg/kg, i.p.), respectively; group V received aspirin
(100 mg/kg i.p.), as a standard drug. The number of muscular contractions was counted over a period of 20 min after acetic acid
injection. The number of writhes in each treated group was compared with control (saline treated group) and the percent inhibition
of the writhes was calculated.
2.6.2. Formalin test
The method used in our study for the assessment of formalininduced flinching behaviour in normal rats was described
previously (Dubuisson and Dennis, 1977; Tjolsen et al., 1992). In
this method, 0.05 ml of formalin (2.5% formaldehyde) was injected
into the plantar surface of the right hind paw, 30 min after treating
the animals with the extracts (50, 100 and 200 mg/kg i.p.). Nociceptive behaviour was quantified as rat walking or can stand on
injected paw; paw partially elevated; total elevation of injected
paw, injected paw licking or biting. Formalin injection induced a
stereotyped response characterized by two well distinct phases;
phase I started almost immediately and was short lasting (0–5 min)
followed, by prolonged tonic phase II lasting (15–30 min). Morphine (3 mg/kg s.c.) was used as a standard drug.
2.6.3. Thermal nociception (hot plate test)
In thermal nociception (hot plate test) mice were screened by
placing them on a hot metal plate maintained at 50 ± 0.05 ◦ C (Dar
et al., 2005). The mice were treated either with vehicle (10 ml/kg
i.p.), PR (50, 100 and 200 mg/kg i.p) or morphine (10 mg/kg s.c.)
an opioid analgesic as a standard drug. Thermal nociception was
estimated by measuring withdrawal response latency in the form
of jumping, withdrawal of the paws or the licking of the paws.
In the pretreatment session, mice were tested on two separated
occasions, each 30 min apart and then only those mice were
selected for the study, which responded within 15 s and which
showed comparatively similar results. The response latencies were
recorded at 0, 30, 60, 90 and 120 min with a cut off period of
30 s to avoid damage to the paw in the absence of response. In
order to investigate the participation of the opioid system in the
antinociceptive effect of PR, naloxone hydrochloride (2 mg/kg s.c.),
a non-selective opioid receptor antagonist was injected, 15 min
prior to the administration of test samples, as explained above and
the hot plate latencies were sequentially measured at 0, 30, 60, 90
and 120 min.
2.7. Diuretic activity
2.5. Acute toxicity studies
The acute toxicity test for PR was carried out to evaluate any
possible toxicity. Swiss albino mice (n = 6) of either sex were tested
by administering different doses of PR by increasing or decreasing
the dose, according to the response of animal (Bruce, 1985). The
dosing patron was 500, 1000 and 2000 mg/kg p.o., while the control
group received only the normal saline. All the groups were observed
for any gross effect or mortality during 24 h.
2.6. Antinociceptive activity
2.6.1. Visceral pain model (acetic acid-induced abdominal
constriction)
The peripheral nociceptive activity was investigated by using
the acetic acid-induced abdominal constriction test (Koster et al.,
The diuretic activity of PR was determined by method previously
established (Jabeen et al., 2009). Male Albino rats were divided in
to four groups (n = 6). The animals were fasted for 24 h and were
fed laboratory diet ad libitum and allowed free access to drinking water. On the day of experiment, the animals of group I was
treated with saline (15 ml/kg p.o) and this group served as control. Similarly, the animals of group II, III and IV were administered
hydrochlorothiazide (10 mg/kg p.o) as standard drug, PR (300 and
600 mg/kg p.o), respectively. The extract was dissolved in saline and
was suitably diluted for administration. Immediately after the drug
treatment, the animals were placed in metabolic cages (1 animal
in each metabolic cage). Urine was collected in graduated cylinders and its volume was recorded at 2, 3 and 6 h. Cumulative urine
excretion was calculated in relation to body weight and expressed
as ml/100 g body weight.
H. Khan et al. / Journal of Ethnopharmacology 127 (2010) 521–527
523
2.8. Phytochemical analysis
2.8.1. Total saponin contents
Saponin contents were determined for the crude extract
and solvent fractions according to the method of Tomowa and
Gjulemetowa (1978). Briefly, 2 g of test samples were taken in a
beaker and 50 ml of petroleum ether was added and heated gently on water-bath to 40 ◦ C for 5 min with regular shaking. The
petroleum ether was filtered and repeated the operation twice with
further 50 ml of pet ether. The marc obtained was extracted with
4× 60 ml of methanol on gentle heating. The methanol layer was
concentrated to approximately 25 ml on water-bath and 150 ml
of dry acetone was added to precipitate the saponins, which was
followed by filtration and drying in oven at 100 ◦ C for constant
weight.
2.8.2. Total alkaloid contents
The total alkaloid contents of PR and subsequent solvent fractions of Polygonatum verticillatum was estimated by using method
developed previously (Obadoni and Ochuko, 2001). Briefly, 2 g of
each was defatted by extraction with petroleum ether, heated gently on water-bath to 40 ◦ C for 5 min with regular shaking. The
marc obtained was acidified with 100 ml of 20% acetic acid in
ethanol and allowed to extract for 4 h. The resulting solution was
filtered, concentrated and then basified with concentrated ammonium hydroxide to pH 9 followed by precipitation. The final weight
of precipitated mass represented the total alkaloid contents.
Fig. 1. Effect of intraperitoneal administration of PR (50, 100, 200 mg/kg) in acetic
acid-induced writhing in mice. Values are reported as mean ± S.E.M. for group of six
animals. The data were analyzed by ANOVA followed by Dunnett’s test. Asterisks
indicated statistically significant values from control. *P < 0.05, **P < 0.01.
3.4. Effect of thermal nociception
2.9. Statistical analysis
Results are presented as the mean ± S.E.M. for at least six animals per group. The statistical analysis of the results was carried out
using one-way ANOVA followed by Dunnet’s multiple comparisons.
The criterion for statistical significance was P < 0.05.
3. Results
As presented in Table 1, PR elicited marked analgesic activity in
the thermal nociceptive test at 50, 100 and 200 mg/kg. The response
was showed by the increase in latency time in seconds. The increase
in latency time was dose-dependent which was measured at 0, 30,
60, 90 and 120 min after the administration of vehicle, PR and standard drug morphine. The peak protection against thermal pain was
achieved at 90 min of PR administration compared to vehicle. The
involvement of opioid receptor was assessed by the administration
3.1. Acute toxicity
No gross behaviour effect or mortality was observed during the
assessment time (24 h), therefore concluded to be safe up to 2 g/kg
p.o.
3.2. Effect of acetic acid-induced abdominal constriction
PR demonstrated a marked and significant (P < 0.01) reduction
(15–72%) in the number of writhes induced by acetic acid. As shown
in Figs. 1 and 2, the analgesic activity at all tested doses (50, 100
and 200 mg/kg i.p.) indicated a dose-dependent relationship in
the inhibition of writhing reflux and the analgesic response of PR
at 200 mg/kg was quite similar to standard drug aspirin (77% at
100 mg/kg i.p.).
3.3. Effect of formalin-induced pain
The formalin-induced flinching behaviour was strongly attenuated by PR in both phases, as compared to control. As shown in
(Fig. 3A and B), the pain relieving potential of the PR was more
prominent and significant (P < 0.01) in the first phase of injected
formalin as compared to second phase. The antinociceptive effect
was found dose-dependent in both phases. Morphine (5 mg/kg s.c.)
offered marked and significant (P < 0.01) reduction in pain induced
by formalin in both the phases.
Fig. 2. Inhibition of writhes in percentage of intraperitoneal administration of PR
(50, 100, 200 mg/kg) in acetic acid-induced writhing in test in mice. Data is expressed
as mean ± S.E.M. (n = 6).
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H. Khan et al. / Journal of Ethnopharmacology 127 (2010) 521–527
Table 1
Effect of the PR on the hot plate (thermal stimuli) test with and without Naloxone in mice at 50, 100 and 200 mg/kg, i.p.
Latency time (mean ± S.E.M.)
Group
Dose mg/kg
Saline
PR
10 mL/kg
50
100
200
10
7.10
7.20
7.20
7.00
7.10
±
±
±
±
±
0.49
0.49
0.35
0.44
0.40
7.20
7.35
7.40
7.40
7.90
±
±
±
±
±
0.13
0.15
0.17
0.13
0.40
7.35
7.65
8.50
11.50
12.70
±
±
±
±
±
0.20
0.15
0.44
0.44**
0.17**
7.40
8.00
9.70
13.70
16.10
±
±
±
±
±
0.42
0.44
0.89
0.84**
0.89**
50
100
200
10
7.30
7.40
7.40
7.60
±
±
±
±
0.44
0.17
0.13
0.26
7.30
7.30
7.10
7.10
±
±
±
±
0.58
0.31
0.89
0.58
7.60
8.20
10.00
7.10
±
±
±
±
0.26
0.44
0.89*
0.40***
7.50
8.10
9.26
6.85
±
±
±
±
0.44
0.53
0.51**
0.60***
0 min
Morphine
With naloxone
PR
Morphine
30 min
60 min
90 min
120 min
7.60
8.00
9.10
12.90
14.36
±
±
±
±
±
0.49
0.44
0.89
0.84**
0.93**
7.50
8.15
9.40
6.85
±
±
±
±
0.89
0.51
1.11*
0.89***
Without naloxone, data was compared with control (Saline 10 ml/kg). With naloxone (2 mg/kg s.c.), was administered 15 min prior to extract or morphine. All the results
were compared with their respective test substances in the absence of naloxone. Values are reported as mean ± S.E.M. for group of six animals. The data were analyzed
by ANOVA followed by Dunnett’s test. Asterisks indicated statistically significant values from control. *P < 0.05, **P < 0.01, ***P < 0.001. The independent t-test was used for
comparison between 2 groups.
of naloxone (2 mg/kg) subcutaneously, 15 min prior to the administration of test samples. The administration of naloxone caused
significant (P < 0.01) attenuation of antinociceptive activity of PR,
as shown in Table 1, establishing the participation of opioid receptor in the central analgesic activity. Moreover, morphine induced
analgesia was completely abolished by the presence of naloxone.
3.5. Effect of diuretic activity
The diuretic activity of PR was evaluated at 300 and 600 mg/kg, is
presented in Fig. 4. The data indicating that PR exhibited mild but
statistically insignificant diuretic activity at 300 mg/kg p.o. compared to control. Surprisingly, the diuretic activity was markedly
decreased at the dose of 600 mg/kg as compared to 300 mg/kg and
was quite similar to normal saline (Fig. 5).
3.6. Effect of total saponins and alkaloids content
The crude extract and subsequent solvent fractions of PR possessed considerable concentration of total saponins and alkaloids
as shown in Figs. 6 and 7.
Fig. 3. Effect of intraperitoneal administration of PR (50,100 and 200 mg/kg) in
formalin-induced flinching behaviour in rats, in the 1st phase (A) and 2nd phase
(B) of formalin test. Values are reported as mean ± S.E.M. for group of six animals.
The data was analyzed by ANOVA followed by Dunnett’s test. Asterisks indicated
statistically significant values from control. *P < 0.05, **P < 0.01.
Fig. 4. Protection (%) of PR (50, 100 and 200 mg/kg i.p.) in the both phases of
formalin-induced pain in rats. Symbols represent mean ± S.E.M. (n = 6).
H. Khan et al. / Journal of Ethnopharmacology 127 (2010) 521–527
525
Fig. 5. Effect of oral administration of PR (300 and 600 mg/kg) in rats as urine volume per 100 kg body weight after 6 h. Hydrochlorothiazide (HCT; 10 mg/kg o.p.)
was standard drug. Values are reported as mean ± S.E.M. for group of six animals.
The data were analyzed by ANOVA followed by Dunnett’s test. Asterisks indicated
statistically significant values from control. *P < 0.05, **P < 0.01.
Fig. 7. Total alkaloids contents in crude extract and subsequent solvent fractions of
rhizomes of Polygonatum verticillatum All. PR-1: crude methanol extract, PR-2: hexane fraction, PR-3: chloroform fraction, PR-4: ethyl acetate fraction, PR-5: butanol
fraction, PR-6: aqueous fraction. Values are expressed as mean ± S.E.M. (n = 3).
4. Discussion
management of different painful conditions, the crude methanol
extract (PR) was screened in various pain models, along with
diuretic activity.
PR showed marked antinociceptive activity in various pain models including visceral pain model, formalin test and hot plate
test. Acetic acid-induced writhing is a highly sensitive and useful test for analgesic drug development especially peripherally
acting analgesics. Acetic acid induces pain by liberating endogenous substances (bradykinin, serotonin, histamine, substance P)
(Tsung-Chun et al., 2007). Hyperalgesia induced by the injection
of acetic acid is characterized by contraction of the abdominal
muscle accompanied by an extension of the forelimbs and body
elongation. These peripheral nociceptive fibers are sensitive to both
narcotics analgesic (morphine) and non-steroid anti-inflammatory
drugs like aspirin (Wibool et al., 2008). In our observation, PR significantly (P < 0.01) reduced the abdominal constriction response
induced by the acetic acid in a dose-dependent manner. Therefore, one possible mechanism of antinociceptive activity of PR
could be due to the blockade of the effect or the release of
endogenous substances (arachidonic acid metabolites) that excite
pain nerve endings by the pharmacologically active principles of
PR.
From a mechanistic point of view, the lack of specificity in
acetic acid-induced writhing test suggesting the involvement of
different nociceptive mechanisms in the reduction of muscular
constriction such as sympathetic system through the release of
biogenic amines, cyclooxygenases and their metabolites inhibition and through opioids receptors mechanisms (Andrade et al.,
2007). This deficiency can be overcome by using other paradigms.
The formalin test is used as a primary behavioural screen for
the assessment of the antinociceptive activity of compounds used
in moderate, long lasting clinical pain. In rats, the spontaneous
nociceptive responses are typically characterized by lifting, biting/licking and flinching of the hind paw after the injection of
formalin into either the dorsal or plantar surface of the paw (Nisar
et al., 2008). Formalin test produced a distinct biphasic response.
First neurogenic phase (0–5 min) occurs within seconds of formalin
injection as a direct result of chemical stimulation of peripherally localized TRPA-1 containing nociceptors (McNamara et al.,
Pain management for patients with chronic pain is a difficult
task because of the risks associated with toxicity due to the drug
intervention (Sam, 2008). Non-steroidal anti-inflammatory drugs
(NSAIDs) are the most widely prescribed medications for the management of painful conditions, but they are frequently causing
gastrointestinal damage (Fiorucci et al., 2001). Ethnobotanicals to
treat diseases is a therapeutic modality, which has stood the test
of time for the treatment of various ailments (Gilani and Atta-urRahman, 2005). To explore new effective and safe analgesic for the
Fig. 6. Total saponins contents in crude extract and subsequent solvent fractions of
rhizomes of Polygonatum verticillatum All. PR-1: crude methanol extract, PR-2: hexane fraction, PR-3: chloroform fraction, PR-4: ethyl acetate fraction, PR-5: butanol
fraction, PR-6: aqueous fraction. Values are expressed as mean ± S.E.M. (n = 3).
526
H. Khan et al. / Journal of Ethnopharmacology 127 (2010) 521–527
2007). The second, later tonic phase (25–30) occurs as a result
of increased primary afferent drive with subsequent sensitization of nociceptive spinal neuron. From a mechanistic point of
view, different analgesics may act differently. Centrally acting drugs
such as opioid inhibit both phases equally but peripherally acting
drugs, such as cyclooxygenase inhibitors (aspirin, indomethacin
and corticosteroids) (Morteza et al., 2004) inhibits only the late
phase.
During our investigation in the formalin test, PR significantly
(P < 0.01) attenuated the hyperalgesia produced by formalin injection in both phases. The pain relieving effect was comparatively
more promising in the late phase and was dose-dependent. Therefore, the antinociceptive activity of PR in formalin test is strongly
attributed to peripherally acting as well as centrally acting pain
mediators. To confirm the participation of central system in the
antinociceptive activity of PR, hot plate test was employed. Hot
plate test is predominantly a spinal reflex and used to test supraspinal analgesia in compounds. It is therefore, selective for centrally
acting analgesic drugs, like morphine, while peripheral antinociceptive agents are found to be inactive on thermal stimulus
(Yu-Feng et al., 2008).
PR exhibited marked inhibition on thermally induced hyperalgesia. The PR possesses significant (P < 0.01) activity 200 mg/kg
after 60, 90 and 120 min of drug administration. Morphine
(10 mg/kg s.c.) being standard drug, showed more potent activity
and was increased with time up to 120 min. Therefore, the result
of our study showed the central mediation in the antinociceptive
activity of PR. For the assessment of opioid system involvement
in the analgesic activity of PR, a non-selective opioid receptor
antagonist, naloxone (2 mg/kg s.c.) was administered. The analgesic activity of PR was significantly (P < 0.001) antagonized by the
administration of naloxone and it is therefore, confirmed the opioid
receptors participation in the attenuation of thermal hyperalgesia.
PR and subsequent solvent fractions of the rhizomes of plant possessed responsible quantity of saponins and alkaloids (Figs. 6 and 7).
The antinociceptive potential of saponins and alkaloids are well
documented in literature (Dilipkumar et al., 2005; Kumar and
Nemmani, 2002; Farouk et al., 2008), therefore, the saponins and/or
alkaloids of PR could be the antinociceptive constituents. Moreover,
to the best of our knowledge on the basis of available literature,
Polygonatum verticillatum is the first member of the genus Polygonatum which demonstrated marked antinociceptive activity in the
present study.
In the traditional system of medicine, PR is used in combination
with other plant based diuretics for the enhancement of urine out
put. Therefore, PR was also screened for the possible diuretic activity. In our assessment on the diuretic activity of PR, mild diuretic
activity was computed at 300 mg/kg compared to saline. In contrast, the urine discharge was similar to saline at 600 mg/kg. The
mild diuretic potential of the plant could be the reason for its use
in polypharmacy for diuresis.
In conclusion, this study demonstrated that the opioid dependant central effect of the PR has synergistic effect by enforcing the
peripheral analgesic effects and thus substantiated the traditional
claim of the plant in the treatment of painful conditions. However,
the diuretic potential of the plant, as a mono-therapy, is not supported by this study. For further clarification of mechanism and
involvement of specific components in the present study, bioassay directed isolation of pharmacologically active molecules of the
plant is needed.
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