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Journal of Medicinal Plants Research Vol. 4(19), pp. 1991-1995, 4 October, 2010 Available online at http://www.academicjournals.org/JMPR Analgesic, antipyretic and anti-inflammatory effects of
Tacca chantrieri Andre
Kittipong Keardrit1, Chaiyong Rujjanawate2* and Duangporn Amornlerdpison3
1Faculty of Science and Technology, Surindra Rajabhat University, Surindra, Thailand. 2School of Health Science, Mae Fah Luang University, Thailand. 3Faculty of Fisheries Technology and Aquatic Resources, Maejo University, Chiang Mai, Tacca chantrieri Andre is an indigenous perennial of the tropics which is used by local healers to
relieve pains of the body and stomach, and as an antidote for food poisoning. The present study was
undertaken to investigate the analgesic, antipyretic and anti-inflammatory activities of T. chantrieri as
claimed in traditional medicine. The ethanolic extract of the plant’s rhizome was prepared and tested in
experimental animals. It was found that the extract significantly inhibited pain caused by acetic acid
injection in the writhing response test in mice and the tail flick test in rats. This finding suggests that
the extract exerts analgesic effect through both peripheral and central mechanisms. The analgesic
effect was not antagonized by pretreatment with naloxone, an opiod antagonist and this signifies a
mechanism other that of the opioid system was utilized. The extract also significantly decreased the
yeast-induced hyperthermia in rats. Anti-inflammatory effect of T. chantrieri extract was demonstrated
in ethylphenylpropiolate-induced ear edema and formalin tests in mice. These findings indicate that the
ethanol extract of T. chantrieri possesses analgesic, antipyretic and anti-inflammatory effects, which is
in accord with its use in traditional medicine.
Key words: Tacca chantrieri Andre, analgesic, anti-pyretic, anti-inflammatory.

INTRODUCTION
Tacca chantrieri Andre (Taccaceae) is an indigenous Therefore, the aim of the current study was to investigate perennial of the tropics. The plant can be ornamental due theses activities of the plant in experimental animals. to its queer looking flower that is shaped like a flying bat. Crude ethanolic extract of T. chantrieri (TCE) was pre- The decoction of T. chantrieri rhizomes, alone or in pared from the plant’s rhizome. The TCE was tested for combination with other herbs, is used by local healers in analgesic activity using writhing responses in mice and South-east Asia to relieve pains of the body and tail flick test in rats. Anti-inflammatory activity was stomach, and as an antidote for food poisoning. Phyto- evaluated by formalin test in and ethyl phenylpropiolate- chemical studies of the rhizomes of T. chantrieri have induced ear edema in rats. Lastly, the TCE was tested for yielded a wide array of saponins and glycosides the antipyretic activity using yeast-induced hyperthermia (Yokosuka et al., 2002; 2005), and some compounds (spirostanol saponins and diarylheptanoid glucosides) However, there has been no pharmacological study to MATERIALS AND METHODS
support the beneficial effect of T. chantrieri. As per the traditional use, we hypothesized that the plant possesses Plant material and extraction
analgesic antipyretic and anti-inflammatory activities. The rhizomes of T. chantrieri were col ected from Payao province in April 2009. The plant was authenticated by one of the authors (Ruj anawate) and the voucher specimen (no. 143) has been deposited at the school of Health Science, Mae Fah Luang *Corresponding author. E-mail: [email protected]. University, Thailand. The air dried powdered rhizome was macerated with 95% ethanol overnight and filtered. The filtrate was (first 5 min) and late phase (last 45 min) with a 10 min lag period in concentrated in vacuo at 55˚C and lyophilized to obtain a dry between phases. Acetyl salicylic acid (ASA) at a dose of 150 mg/kg ethanolic extract (15.5% yield) which from now on is referred as TCE. The TCE was subsequently reconstituted in water to required Ethyl phenylpropiolate-induced ear edema in rats
The method was modified from that of Brattsand et al. (1982). Male rats weighing 30 to 50 g were used. Ear edema was induced by Male Sprague-Dawley rats weighing 150 - 200 g (or stated other- topical application of ethyl phenylpropiolate (EPP) dissolved in wise) and Swiss albino mice weighing 25 to 30 g were purchased acetone (50 mg/ml) in a volume of 10 µl to the inner and outer from the National Laboratory Animal Center, Salaya Mahidol surfaces of both ears (20 µl/ear). The test sample was topical y University, Thailand. They were acclimatized for at least 7 days in applied to the ear just before the irritant. The thickness of each ear an animal room where the temperature was maintained at 22 ± 3˚C was measured with a vernier caliper at 15, 30, 60 and 120 min after and there was a 12 h light-dark cycle. The food was supplied by the edema induction. The edema thickness of the sample-tested Pokphan Animal Feed Co. Ltd. Bangkok. The bedding was group was compared to that of the vehicle group using autoclaved. The animals had free access to food and water. Al phenylbutazone (PHBZ) at a dose of 1 mg/ear as a positive control. animals received humane care in compliance with the ethics in the use of animals issued by the National Research Council of Thailand Yeast-induced hyperthermia in rats
Rats were restrained individual y in a plastic cage. A probe (model Writhing responses in mice
it-rr4) of an EXACON electronic thermometer (model MC8940, EXACON Scientific Instrument Aps, Denmark) was inserted to a This is a basic screening test for analgesic activity. Briefly, a typical depth of 2 cm into the rectum in order to record the initial rectum writhing response in mice was produced by an intraperitoneal temperature. The animals were then fevered by injection of 20% injection of 0.75% acetic acid aqueous solution at a dose of 0.1 suspension of Brewer’s yeast in 0.9% NaCl at a dose of 10 ml/kg ml/10 g body weight. The TCE was administered intraperitoneal y at subcutaneously in the back below the neck. Eighteen hours later, 30 min before the acetic acid injection (Nakamura et al., 1986). the rectal temperatures were recorded again. Those animals with Indomethacin and morphine at a dose of 10 mg/kg were served as rectal temperatures lower than 38.0˚C were excluded from the positive controls for local y and central y acting, respectively. Five experiment. The TCE was administered intraperitoneal y and the minutes later, the number of writhes was counted over a period of rectal temperature was then again recorded every 30 min for 2 h. 15 min. In the other two groups, mice were pretreated with an opiod antagonist naloxone (5 mg/kg, i.p.), 15 min before administration of the TCE (500 mg/kg, i.p.) or morphine (10 mg/kg i.p.). Statistical analysis
Data were subjected to statistical analysis using ANOVA and Tail flick test in rats
statistical comparison was done using Duncan Multiple Range Test. The value exceeding 95% confidence limits was considered to be The response to thermal pain was evaluated according to the tail flick test described by D’Amour and Smith (1941). The rat was placed on the tail-flick unit (Ugo Basile), so that the tail occluded a slit over a photocel . Heat was applied by a 100-W lamp mounted in a reflector. The apparatus was arranged so that when the operator turned on the lamp a timer was activated. When the rat felt pain and flicked its tail, light fel on the photocel then the timer was Given intraperitoneal y, TCE at doses of 250 and 500 automatical y stopped. The light intensity was adjusted to give a mg/kg significantly inhibited acetic acid-induced writhing normal reaction time of 2 to 4 s. A 10 s cut-off time was used in responses as shown in Table 1, which also shows the order to prevent tissue damage. Two control readings, taken 30 min similar effect of reference compounds, indomethacin and apart, were averaged and constituted the control reaction time. The morphine. Pretreatment of the animals with naloxone extract was administered (i.p.) immediately after this step, and 30 could significantly reverse the analgesic effect of min later, the post-extract reaction time was measured. The analgesic response was calculated as a percentage of the morphine but not that of the TCE. Table 2 shows that maximum possible response time. Morphine at a dose of 10 mg/kg TCE at a dose of 50 mg/kg could not significantly inhibit the pain response in the tail flick test while at higher doses (125, 250 and 500 mg/kg) the extract significantly prolonged the reaction time as did morphine, the positive Formalin test in rats
The formalin test comprised the early phase and the late phase In the formalin test, intraperitoneal administration of assessment of the analgesic effect which were performed TCE at doses of 125, 250 and 500 mg/kg significantly separately according to the method of Dubuisson and Dennis lowered the number of flinches in both the early and late (1977). Rats were injected intraperitoneal y with the TCE. Thirty phases while TCE at a dose of 50 mg/kg could minutes later, they were administered 50 µl of a 2.5% solution of significantly lowered the number of flinches only in the formalin, subcutaneously under the plantar surface of the left hind paw. They were then placed in an observation chamber and late phase (Table 3). TCE showed significant inhibition in monitored for 1 h. The number of flinches indicated the severity of the EPP-induced rat ear edema model as shown in Table pain. Analgesic effect was determined in two phases: early phase 4. The information obtained also indicated that TCE is Table 1. Effect of Tacca chantrieri ethanolic extract (TCE) on acetic acid-induced writhing
Treatment (i.p)
No. of writhes
Inhibition (%)
Data are expressed as mean ± S.E.M. (n = 8). ∗significant different from control (p < 0.05). Table 2. Effect of Tacca chantrieri ethanolic extract (TCE) on radiant heat-induced tail flick in rats.
Treatment (i.p.)
Inhibition (%)
Data are expressed as mean + S.E.M. (n = 10). Tc = control reaction time; Tr = reaction time after injection of test drugs. ∗ Significantly different from Tc (p < 0.05). markedly more potent than phenylbutazone (PHBZ), the analgesics such as morphine. In contrast, the late phase reference drug in this test. In the yeast-induced may be due to an inflammatory response partly mediated hyperthermia in rats, it was found that TCE at doses of by prostaglandins and can be inhibited by NSAIDs and 125, 250 and 500 mg/kg could produce significant anti- steroids, as wel as by the central y acting drugs. As inflammation occurs at the site of formalin injection, it is possible to elucidate the role of inflammation on the responses in the two phases. It was previously shown DISCUSSION
that ASA and indomethacin are anti-nociceptive through partial y different modes of action in this test (Hunskaar et The analgesic effect of the CAF was evaluated using the acetic acid-induced writhing, the tail flick and the formalin It was also shown that ASA does not have any delay of (early and late phases) tests. These procedures are used onset of its action in the early phase compared to to detect central and peripheral analgesia and to morphine (Hunskaar and Hole, 1987). Both of these distinguish analgesic from anti-inflammatory properties. studies suggested that ASA has some effects which The writhing test is used for screening of the analgesic cannot be attributed to the inhibition of prostaglandin activity regardless of the central or peripheral causes. In the tail flick test, which uses a thermal stimulus, an In this study, the analgesic effect of the extract was increase in reaction time is general y considered as an demonstrated in the writhing response in mice and tail important parameter of central analgesic activity (Chang flick tests in rats. The TCE at doses of 250 and 500 and Lewis, 1989). The formalin test (Hunskaar et al., mg/kg was active in both the writhing response and the 1985) is sensitive to NSAIDs and other mild analgesics. tail flick tests. This finding suggests that TCE exerts The test employs a chemical nociceptive stimulus that analgesic effect through both peripheral and central elicits a spontaneous response indicative of pain. The mechanisms. The finding that pretreatment of the mice test has two different phases, possibly reflecting different with an opiod antagonist naloxone could not reverse the types of pain (Dubuisson and Dennis, 1977; Hunskaar et analgesic effect of the TCE in acetic acid-induced pain al., 1985). The early phase may be due to direct effects on nociceptors and can be inhibited by central y acting Table 3. Effect of Tacca chantrieri ethanolic extract (TCE) on the formalin test in
No. of flinches
Treatment (i.p.)
Data are expressed as mean ± S.E.M. (n = 8). ∗ Significantly different from control (p Table 4. Effect of topical application of Tacca chantrieri ethanolic extract (TCE) on EPP-induced ear oedema in rats.
edema ear thickness (µm)
% inhibition
Treatment
12 ± 4∗ 105 ± 16∗ 130 ± 18∗ 142 ± 14∗ 21 ± 3∗ 98 ± 10∗ 120 ± 17∗ 134 ± 10∗ Data are expressed as mean ± S.E.M. (n = 10). ∗ Significantly different from control (p < 0.05); PHBZ = phenylbutazone. Table 5. Effect of Tacca chantrieri ethanolic extract (TCE) on yeast-induced hyperthermia in rats.
Rectal temperature (˚C)
Treatment
(mg/kg i.p.)
18 h after yeast
injection
Minutes after TCE injection
Data are expressed as mean + S.E.M. (n = 10). ∗Significantly different from control (p < 0.05). formalin test in mice was conducted to distinguish and antipyretic activities of the TCE. In conclusion, this analgesic from anti-inflammatory properties (Hunskaar et study confirms the ethnomedical use of T. chantrieri as al., 1985). The finding that TCE as wel as ASA exerted a an analgesic, antipyretic and anti-inflammatory agent. It’s marked analgesic activity in the late phase of the formalin worth noting that steroidal saponins have been isolated test suggests an effect on acute inflammation. Results from the rhizomes of the plant (Yokosuka et al., 2002; from the ear edema test using EPP as an inducer of 2005) but it is not clear at present whether these substances are responsible for the pharmacological Final y, the antipyretic effect of the extract was demonstrated in yeast-induced hyperthermia in rats. The production of prostaglandins appears to be a final com- mon pathway responsible for fever production induced by ACKNOWLEDGEMENTS
several pyrogens (Milton, 1982). Therefore, it is reasonable to assume that the inhibition of prostaglandin This work was financial y supported by Thailand biosynthesis may take part in the anti- inflammatory Luang University (grant no. 52107030011). Hunskaar S, Fasmer OB, Hole K (1985). Formalin test in mice, a useful technique in evaluating mild analgesics, J. Neurosci. Methods, 14: Hunskaar S, Berge OG, Hole K (1986). Dissociation between REFERENCES
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