Antinociceptive activities of aqueous and ethanolic extracts of Alpinia calcarata rhizomes in rats

doi:10.1016/j.jep.2004.07.015

Journal of Ethnopharmacology

Volume 95, Issues 2–3, December 2004, Pages 311-316

https://doi.org/10.1016/j.jep.2004.07.015 Get rights and content

Abstract

Rhizomes of Alpinia calcarata Roscoe (Zingiberaceae) posses several bio-activities and are used in traditional medicine of Sri Lanka. However, their antinociceptive activity has not been investigated so far. The aim of this study therefore, was to examine the antinociceptive activity of hot water extract (HWE) and hot ethanol extract (HEE) of Alpinia calcarata rhizomes using rats and three models of nociception (tail flick, hot plate and formalin tests). Different concentrations of HWE (100, 250, 500, 750, 1000 mg/kg) and HEE (100, 250, 500, 750, 1000 mg/kg) were made and orally administrated to rats and the reaction times determined. The results showed that the extracts have marked dose-dependent antinociceptive activity, when evaluated in the hot plate and the formalin tests but not in the tail flick test. The antinociceptive effect was slightly higher in HEE than that of HWE. The antinociceptive effect was mediated via opioid mechanisms.

Introduction

Alpinia calcarata Roscoe (Zingiberaceae), Heen-araththa in Sinhala, Amkolinji in Tamil and Kattuchena in Malay is a rhizomatous perennial herb. The mature rhizomes are branched and dense with a light to dark brown color. The leaf of the plant is simple, alternative, 25–32 cm long, 2.5–5 cm broad. The flowers are irregular, bisexual and pendanculate. Terminal densed flowers are found in panicles 8.5 cm long (Dassanayake and Fosberg, 1981, Jayaweera, 1982). Alpinia calcarata is cultivated in tropical countries including Sri Lanka, India and Malaysia. Especially the rhizomes of Alpinia calcarata are used for the medicinal purposes (Jayaweera, 1982).

Kong et al., 2000, Kong et al., 2002 have isolated some diterpenes such as calcaratarins A–E, sesquiterpenes such as shyobunone and coumarins such as herniarin from the rhizomes of Alpinia calcarata grown in China. On the other hand, some benzenoids such as protocatechuic acid, vanillic acid and syringic acid, flavonoids and alkaloids were isolated from the leaves of Alpinia calcarata grown in India (Merh et al., 1986). In our studies (Arambewela et al., in press) we have isolated 18 volatile constituents in essential oils of Sri Lankan grown Alpinia calcarata rhizomes, roots and leaves. 1,8-Cineol was found to be the major constituent in the oils of rhizomes and leaves while in the roots, it was α-fenchyl acetate.

Indian researches have shown antibacterial (George and Pandalai, 1949), antifungal (Pushpangadan and Atal, 1984) and anthelmintic activity (Kaleysa, 1975) in extracts of Alpinia calcarata. However, in Indian traditional ayurveda medicine Alpinia calcarata is not generally used. On the other hand, in Sri Lankan traditional ayurveda medicine rhizome of Alpinia calcarata is recommended as an aphrodisiac and a decoction is widely used in the treatment of bronchitis, cough, respiratory ailments, diabetics, asthma (Ramanayake, 1994) and arthritis (Ramanayake, 1994, Arambewela et al., 1995). Generally drugs that are used for arthrities have antinociceptive and anti-inflammatory properties. However, antinociceptive activity of Alpinia calcarata rhizomes has not been investigated using scientifically controlled experiments. Therefore, this study was undertaken to examine whether extracts of rhizomes of Alpinia calcarata possess antinociceptive activity. This was tested in rats using oral administration of hot water and hot ethanolic extracts.

Section snippets

Plant material

Fresh Alpinia calcarata rhizomes were collected from home gardens in Kaluthara district, Sri Lanka between August and November 2002. The plant material was identified and authenticated by Mr. D.H.P. Peramunagama, Curator of National Herbarium, Royal Botanical Gardens, Peradeniya, Sri Lanka. A voucher specimen (AS 01) was deposited in the Industrial Technology Institute, Colombo 7, Sri Lanka.

Animals

Healthy adult cross-bred male albino rats (weighing 200–250 g) were used throughout the experiment. They

Phytochemical screening of HWE and HEE

Phytochemical screening revealed the presence of alkaloids, steroids, coumarins, reducing sugars and flavonoids in both extracts (results not shown).

Hot plate and tail flick tests

In the hot plate test, significant prolongation of the reaction time was evident with both extracts at 1 h post-treatment (Fig. 1, Fig. 2). All doses of HWE significantly prolonged (P < 0.05) the reaction time for up to 4 h apart from the 1000 mg/kg dose which was only active for 1 h. On the other hand, all the tested doses of HEE significantly

Discussion

The results demonstrate that extracts of rhizomes of Alpinia calcarata have antinociceptive activity as evaluated in the hot plate test and not in the tail flick test. This indicates centrally mediated antinociceptive activity of the plant extracts against the acute pain: both hot plate and tail flick tests measure centrally mediated transient pain (Lopez-Munoz et al., 1993). It is claimed that the antinociceptive activity of extracts was genuine as there was no change in the retain times of

Acknowledgements

The authors express their gratitude to Mr. J.R.A.C. Jayakody, University of Colombo, Department of Zoology, for assisting in this study and National Science Foundation for the Research grant (SIDA (1L) 2000/BT/03).

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Ethnogynaecology is an emerging branch of science dealing with the treatment of gynaecological ailments by tribals, local healers, and traditional practitioners. The ethnogynaecological importance of medicinal plants in India is a fertile area to conduct more scientific studies to evaluate their potentialities, to isolate bioactive compounds, and thereby to develop drugs for the common gynaecological health-related issues faced by women everywhere.
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The detailed investigation of ethnobotanical and ethnogynaecological-related literature published between 1985 and 2021 by different scientific tools such as journals, books, and current electronic databases like Springer Link, SciFinder, Google Scholar, Web of Science, Wiley, ACS, Science Direct and Pubmed have been considered for the present study. The study included 300 articles published between 1985 and 2021 by scientific search using various standard databases. The tribals, vaidyas, traditional practitioners, indigenous medical healers, and local people of different regions in India have recognized the importance of ethnogynaecological uses of plants. The study on ethnogynaecology is limited to a few common but significant gynaecological issues including abortion, contraception, infertility, menstruation, leucorrhoea, and obstetrics. The phytocompound compounds isolated from various parts of the plants and responsibility for the gynaecological action were documented.
The major ethnogynaecological disorders recorded by various studies are leucorrhoea, abortion, contraceptives, infertility and related issues, and obstetrics including the irregular physiological process of menstruation. The ethnogynaecological and ethnobotanical information has been recorded from almost all the states of India; the highest number of records on ethnogynaecology was reported from the state of Madhya Pradesh. The most explored tribal populations to record ethnogynaecological knowledge belong to the following tribes: Bhil, Munda, Irula, Kani, Malayali, Meena, Paliyar, Muthuvar, Oraon, Narikuravar, Mannan, Malayarayan, and Malapandaram. Moreover, limited or no study has been attempted to prove the knowledge of ethnogynaecology of these tribes and the efficiency of their crude drugs against pharmacological actions. The paste prepared from various parts of the plants has been used widely as primary health care materials for abortion, obstetrics, menstruation, female infertility and male infertility. Phenols, glucoside, steroids and fatty acids reported with cytotoxic activities are connected to several gynaecological disorders whereas flavonoid, coumarin, sitosterol disrupt pregnancy. The phenolic compounds induced spontaneous abortion due to the major composition aristolochic acid, ceryl alcohol, β-sitosterol. Coreopsin, butin, isobutrin, monospermoside, palastrin, butrin. Mucunine, lecithin, prurieninine, gluthione and luteolin, Indicine, kaempferol, apigenin and quercetin effected therapeutic activity against leucorrhoea. Lignin, friedelin and beta-sitosterol are reported with abortifacient properties and therapeutic ability for leucorrhoea and menorrhagia. Tannins, mimusopsic acids, taraxerol and spinaserol effected fertility problems in women and tannins, saponins, flavonoids, steroids, terpenoids and alkaloids which effected infertility.
This review reported comprehensive data on ethnogynaecological knowledge published from available literature and evident that the indigenous medical system of Indian tribes has also contributed considerably to the healthcare system and drug development of India. The fresh plant parts were identified as effective materials against various gynaecological illnesses including infertility. The root is considered an excellent plant part against obstetrics followed by abortion, menstruation, and leucorrhoea. These studies need experimental proof as well as standardization to confirm their efficiency. Promoting the sustainable use and the equitable sharing of benefits to the knowledge provider is a pathway for harnessing the conservation of this knowledge.
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At present information regarding traditional medicine is a fast step for the development of modern medicine. Alpinia purpurata is the medicinal plant which belongs to the family of Zingiberaceae, it constituents Antidermatophytic [16], antinociceptive [17], hepatoprotective [18], immunostimulatory [19], and anticancer [20] activities. In order to validate these new therapies, animal models are needed.
Numerous Chinese herbal extracts have been proved to inhibit the development of prostate cancer. Zingiberaceae family is one type of plant used as an ingredient in traditional Chinese medicine (TCM). The genus Alpinia species of the Zingiberaceae family are well-known traditional herbal medicine for anti-inflammatory, antioxidant, antimicrobial, anti-dermatophytic, anti-nociceptive, hepatoprotective, immune stimulatory and anticancer activities. In vivo anticancer activity of Alpinia purpurata were studied in MNU and testosterone induced prostate cancer rats.
The induction of prostate cancer by using Wistar strain rats. The animals were divided into five groups. Group 1- Control, normal healthy rats Group 2- Rats were induced prostate cancer by daily intraperitoneal injection of testosterone (T) (100 mg/kg bw) in 0.3 ml propylene glycol for 3 days. One day after the testosterone injection, all the rats received intravenous injection of N-methyl N-nitrosourea (MNU) (50 mg/kg bw) (dissolved in saline at 10 mg/ml), through the tail vein for a period of one week. One week after MNU administration, rats received daily intraperitoneal injection of 2 mg/kg bw testosterone for 60 days. Group 3 - Rats were treated as group 2 along with Finasteride (25 mg/kg bw) supplemented throughout the entire experimental period of 2 months. Group 4 - Rats were treated as group 2 and simultaneously treated with daily dose of 200 mg/kg bw of the crude leaf ethyl acetate extract of Alpinia purpurata through oral gavage for 2 months. Group 5 - Rats were received daily dose of 200 mg/kg bw of crude, leaf ethyl acetate extract of Alpinia purpurata through oral gavage for 2 months. After the experimental period of 4 months, the animals were sacrificed by cervical dislocation under mild chloroform anesthesia. All biochemical parameters and histopathological studies were carried out.
The biochemical parameters such as total protein, enzymatic and non-enzymatic antioxidants and membrane bound enzymes were decreased whereas LPO, tumor markers, nucleic acid constituents were increased in prostate and seminal vesicle tissues of cancer induced group. The hematological, lipid parameters were altered and prostate markers, renal markers and liver markers were increased in serum. Those levels were brought back to near normal upon treatment with ethyl acetate leaf extract of Alpinia purpurata and standard drug finasteride. No significant changes were observed on treatment with plant extract alone group which indicated that there is no toxic substance present in Alpinia purpurata which was confirmed by the histopathological analysis of organs liver, kidney, prostate gland and seminal vesicle.
On the basis of our findings, it is concluded that Alpinia purpurata might be used as an anticancer agent for the management of carcinoma in future.
Bioassay-guided fractionation of a dried commercial source, Alpinia galanga (L.) Willd rhizomes extract, against Culex pipiens (Diptera: Culicidae)
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Rhizomes of the Zingiberaceae family are used in traditional recipes in many ancient cultures for the treatment of various diseases (Chhabra et al., 1993; Meng et al., 2005). The plant possesses antimicrobial (Janssen and Scheffer, 1985), anti-amoebic (Sawangjaroen et al., 2006), anticancer (Samarghandian et al., 2014; Banjerdpongchai et al., 2011), anti-inflammatory, analgesic (Phitak et al., 2009), antiulcer (Yang, 1999), platelet-activating factor inhibitory (Jantan et al., 2005), antinociceptive (Bendjeddou et al., 2003), antioxidant (Juntachote and Berghofer, 2005), and hepatoprotective (Arambewela et al., 2004) activities. Several compounds have been isolated from the rhizome of A. galanga, including β-sitosterol diglucosyl caprate (Jaju et al., 2010), galangoisoflavonoid (Jaju et al., 2009), methyl eugenol, p-coumaryl diacetate, trans-p-acetoxycinnamyl alcohol, 1′-acetoxyeugenol acetate, trans-3,4-dimethoxycinnamyl alcohol, p-hydroxycinnamaldehyde, p-hydroxybenzaldehyde, trans-p-coumaryl alcohol, galangin, galanganol B, trans-p-coumaric acid (Kaur et al., 2010), hydroxychavicol acetate, and acetoxychavicol acetate (Min et al., 2009).
The increasing risk of insecticide resistance in mosquito populations has led to the search for new larvicidal agents. Evaluation of bioassay-guided fractionation of the rhizome extract of Alpinia galanga (L.) Willd against Aedes aegypti was assessed. Bioactive fractions were isolated from the rhizome extract of A. galanga using a Soxhlet extractor and chromatography techniques, and subsequently tested against the fourth instar of Culex pipiens. The lethal concentration (LC) was calculated via log-probit analysis. The active fraction was evaluated by gas chromatography-mass spectroscopy (GC–MS) and infrared (IR) analysis. The highest larvicidal potential obtained from bioassays using the Soxhlet apparatus was observed in dichloromethane (DCM) and ethyl acetate (EtoAc) extracts, with LC₅₀ values of 124.49 and 176.30 ppm, respectively, after 24 h of exposure. Subsequently, the DCM extract was subjected to column and thin-layer chromatography. Results of the DCM extraction and the active TLC fraction (F133) of the Rf value 0.5 revealed that LC₅₀ and LC₉₀ values decreased over time. The F133 fraction of A. galanga exhibited zero hatchability (100% mortality) at LC₅₀ (63.416 ppm) and LC₂₅ (31.70 ppm) against Cx. pipiens eggs. GC–MS analysis of the active thin-layer chromatography TLC fraction (F133) revealed the presence of phenol 2 4-bis (1 1-dimethylethyl), which was identified as the major compound. Alpinia galanga extract is a promising candidate for the control of mosquito populations. Further study is required to determine the effect of the extracts on non-target organisms.
Alpinia calcarata Roscoe: A potent antiinflammatory agent
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Opioid receptor-mediated activities can suppress the “inflammation” in the carrageenin induced paw oedema (Planas et al., 2000). Alpinia calcarata extracts also exhibited a potent antinociceptive activity via opioid receptor mediators (Arambewela et al., 2004). Therefore, opioid receptor mediators also play an important role in inducing antiinflammatory activity of Alpinia calcarata extracts.
Alpinia calcarata Roscoe (Family: Zingiberaceae) rhizomes are often used in Sri Lankan traditional systems of medicine as a remedy for bronchitis, cough, respiratory ailments, diabetics, asthma and arthritis. Generally drugs that are used for arthritis have antinociceptive and antiinflammatory properties. However, validity of the antiinflammatory activity has not been scientifically investigated so far. Therefore, the aim of this study was to investigate the antiinflammatory potential of Alpinia calcarata rhizomes using hot water extract (AWE) and hot ethanolic extract (AEE).
The antiinflammatory activity of Alpinia calcarata was evaluated by use of the carrageenan-induced paw oedema model in rats. In addition, the mechanism/s by which Alpinia calcarata is mediated the antinflammatory activity was assessed by determining its effects on (a) membrane stabilizing, (b) antihistamine and (c) prostaglandin synthesis inhibition activity.
All the tested doses of AWE and AEE (250, 500, 750, and 1000 mg/kg) produced a significant (P ≤ 0.05) inhibition of the inflammation, most pronounced at 4 h after the injection of carrageenan. The antiinflammatory effect induced by 500 mg/kg of AEE was superior than the reference drug, indomethacin at 4 h. Inhibition of histamine and prostaglandin synthesis production is probable mechanisms by which Alpinia calcarata mediates its antiinflammatory action.
These findings rationalize the traditional usage of Alpinia calcarata as an antiinflammatory agent for the first time.
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To explore cytotoxic activity of ethanol extract of Alpinia calcarata Rosc (EEAC) rhizome against Ehrlich ascites carcinoma (EAC) tumor bearing Swiss Albino mice.
In the present study, its anti-neoplastic activity has been studied by monitoring parameters like tumor weight measurement, survival time, tumor cell growth inhibition, haematological characteristics etc.
It was found that EEAC at dose 8 mg/kg/day (i.p.) significantly decreased tumor weight (62.0%; P <0.01), increased life span (70.25%; P <0.01) and reduced tumor cell growth rate (85.7%; P <0.01) in comparison to those of EAC bearing mice. The plant extract also improved the depleted haematological parameters like RBC, WBC, Hb%, differential counts (e.g. lymphocytes, neutrophils, monocytes etc) of EAC bearing mice towards normal. The host toxic effects were not very high and recovered gradually towards normal within a few days after treatment.
EEAC exhibits potent in vivo cytotoxic activity against EAC tumor bearing Swiss Albino mice. So, the plant can be considered as a probable new source of antitumor agents.
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Antinociceptive activities of aqueous and ethanolic extracts of Alpinia calcarata rhizomes in rats

Abstract

Introduction

Section snippets

Plant material

Animals

Phytochemical screening of HWE and HEE

Hot plate and tail flick tests

Discussion

Acknowledgements

Journal of Ethnopharmacology

Journal of Pharmacological Methods

Neuroscience Letters

Journal of Ethnopharmacology

Pain

Traditional Treatment in Sri Lanka for Chronic Arthritis

Antinociceptive effects of the alkaloid epibatidine: further studies on involvement of nicotine receptors

Drug Developing Research

A Revised Hand Book to the Flora of Ceylon

The formalin test: a quantitative study of the analgesic effect of morphine, meperidine and brain stem stimulation in rats and cats

Pain

Biological and phytochemical screening of plants

Journal of Pharmaceutical Science

Validity of head dipping as a measure of exploration in modified hole-board

Psychopharmacology

Investigations on plant antibiotics

Indian Journal of Medicinal Research

Effects of an intrathecal dopamine agonists, apomorphine, on thermal and chemical evoked noxious responses in rats

Brain Research

Screening of indigenous plants for anthelmintic action against human Ascaris lumbricoides

Indian Journal of Physiological Pharmacology