Antinociceptive activity of the methanolic extract of Kaempferia galanga Linn. in experimental animals
Introduction
Medicinal plants are important sources of new chemical substances that potentially have strong therapeutic effects. Most people living in developing countries are almost completely dependent on traditional medical practices for their primary health care needs and higher plants are known to be the main source for drug therapy in traditional medicine (Calixto, 2005). Kaempferia galanga Linn. (Zingiberaceae), commonly called “Proh Hom” in Thai, is an acaulescent perennial growing in Southern China, Indochina, Malaysia, India and Thailand. This medicinal plant has been extensively used for treatment of various disorders including hypertension, rheumatism and asthma (Zakaria and Mustafa, 1994). In Thailand, the rhizome of Kaempferia galanga is used by people in many regions for relieving toothache, abdominal pain, muscular swelling and rheumatism (Sirirugsa, 1997). In addition, the ethnomedical uses of aerial part of Kaempferia galanga are claimed in Thai traditional text books; the stem is used for menstrual stimulation and in the treatment of dyspepsia, whereas leave and flower are used for the treatment of Tinea versicolor, and eye diseases and seizures, respectively (Pongboonrod, 1976, Thamaree and Tankeyoon, 1981, Sighabutra, 1993). The most common indications for its use, besides hypertension, include rheumatism, asthma, headaches, cough, toothaches and as a poultice for applying to bruises and wounds (Perry and Metzger, 1980). In Malaysia and Indonesia, this plant is used to make a gargle, the leaves and rhizomes are chewed to treat coughs, or pounded and used in poultices or lotions applied to relieve many ailments; the juice of the rhizome is used as an expectorant and carminative, and is often a part of children's medicine and tonics (Othman et al., 2006). The rhizome is also used to treat abdominal pain, and as an embrocation or sudorific to treat swelling and muscular rheumatism (Othman et al., 2006). In China, this plant is used as a remedy for toothache, as a stimulant, carminative to treat cholera, and to treat contusions, chest pains, headache and constipation (Ibrahim and Rahman, 1988, Mustafa et al., 1996). Additionally, the rhizomes of the plant that contains essential oils have been used in a decoction or powder for indigestion, cold, pectoral and abdominal pains, headache and toothache (Kanjanapothi et al., 2004). Its alcoholic maceration has also been applied as a liniment for rheumatism (Keys, 1976, Lieu, 1990). Some pharmacological activities of Kaempferia galanga have been reported, for example, as a smooth muscle relaxant and vasorelaxant (Mustafa et al., 1996, Othman et al., 2006).
Kaempferia galanga possesses several bioactivities and is widely used in Thai and other Asian traditional medicine but so far its antinociceptive activity has not been investigated. Phytochemical studies carried out with Kaempferia galanga revealed many different volatile oils. The major chemical constituents of the volatile oil extracted from dried rhizome were ethyl-p-methoxycinnamate (31.77%), methylcinnamate (23.23%), carvone (11.13%), eucalyptol (9.59%) and pentadecane (6.41%), respectively (Tewtrakul et al., 2005). Other constituents of the rhizome include cineol, borneol, 3-carene, camphene, kaempferal, cinnamaldehyde, p-methoxycinnamic acid, ethyl cinnamate and ethyl p-methoxycinnamate (Nakao and Shibu, 1924). A methanolic extract of the rhizome contains ethyl p-methoxy-trans-cinnamate, which is highly cytotoxic to HeLa cells (Kosuge et al., 1985).
In the present study, Kaempferia galanga was selected because it is one of the medicinal plants commonly used in remedies to treat abdominal pain, toothache, swelling and rheumatism in Thai traditional medicine and other countries in Southeast Asia. However, up to date no ethnopharmacological data have previously been systematically conducted to evaluate the antinociceptive action supporting traditional uses of this plant in folklore medicine. In this work we evaluate the “Proh Hom” antinociceptive activity in experimental animals using methanol extract. The reason to use methanol extract in this investigation is that methanol is more nonpolar than water, therefore, several substances including volatile oils, the major chemical constituents of Kaempferia galanga, would be expected to be more soluble in methanol fraction than in water extract.
Section snippets
Plant material
Kaempferia galanga were collected in January 2006 from the Chana District, Songkhla Province, Thailand. The plant was identified by Prof. Puangpen Sirirugsa, Department of Biology, Faculty of Science, Prince of Songkla University. A voucher specimen no. 200601 of the plant material has been deposited in the Prince of Songkla University Herbarium, Department of Biology, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, Thailand.
Experimental animals
Male Swiss albino mice and Wistar rats were used
Acetic acid-induced writhings in mice
The MEKG administered orally at different doses (50, 100 and 200 mg/kg) caused a significant inhibition (42.75%, 59.57% and 70.60%, respectively) compared to the control (p < 0.01) of the writhing responses induced by acetic acid (0.6% v/v, i.p.). The decrease in the number of writhings was dose-dependent. Aspirin (100 mg/kg, p.o.) produced a 51.06% reduction compared to the control. The results are provided in Table 1.
Formalin test in mice
MEKG at doses of 50, 100 and 200 mg/kg, p.o. had a significant dose-dependent
Discussion
In the present study, our results demonstrated that MEKG possesses significant antinociceptive activity as evaluated in the acetic acid-induced writhings, formalin test, hot plate test and tail-flick test. The acetic acid-induced writhing model is a chemical stimulus widely used for the evaluation of peripheral antinociceptive activity (Gene et al., 1998). In this model, pain is generated indirectly via endogenous mediators like bradykinin, serotonin, histamine, substance P and prostaglandins,
Conclusion
The MEKG exhibits antinociceptive activity. The proposed mechanisms of antinociceptive activity based on the pain models used in this study show that they are likely to be mediated peripherally and centrally (spinally and supraspinally) on the nervous system. In addition, the antinociceptive effect of the extract was abolished by naloxone in the same manner as for morphine both in the hot plate and tail-flick tests, indicating that the extract acts partly through opioid-mediated mechanisms. The
Acknowledgements
This research project was supported by grants from Prince of Songkla University (Contract No. SCI50200200180S), Graduate School of Prince of Songkla University, Hat Yai, Songkhla 90112, Thailand. Special thanks are extended to Dr. Brian Hodgson for his excellent reading and corrections to this manuscript.
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