Antinociceptive activity studies in mice with methanol extracts of two mushroom species: Lentinula edodes (Marasmiaceae) and Ganoderma lucidum (Ganodermataceae).
Lentinula edodes (Berk.) Pegler (Marasmiaceae) is commonly known as the shiitake mushroom and is prized throughout the world as a culinary item. It is native to East Asia, where it is also considered a mushroom with medicinal properties. Ganoderma lucidum (Curtis) P. Karst. (Ganodermataceae), commonly known as the reishi mushroom, is also edible. It is a polypore mushroom and has a red-varnished, kidneyshaped cap. This mushroom species is known to have been used as a medicinal mushroom in China for over 2,000 years.
Mushrooms in general are known for their various pharmacological effects, and the above two mushroom species are no exceptions. An exo-polymer produced from a submerged mycelial culture of Lentinula edodes reportedly demonstrated hypoglycemic properties through reduction of sugar levels in the plasma, and increasing concentrations of plasma insulin, when administered to streptozotocin-induced diabetic rats (Yang et al., 2002). Oral ingestion of mycelia extract of this mushroom species reportedly inhibited B16 melanoma growth in C57BL/6 mice (Tanaka et al., 2011). Shiitake mushrooms have also been shown to reduce the incidence and severity of collagen-induced arthritis in dilute brown non-agouti mice (Chandra et al., 2011). Antioxidant activity and hepatoprotective effects of shiitake mushroom against paracetamol-induced hepatotoxicity have been reported (Sasidharan et al., 2010).
Polysaccharides obtained from Ganoderma lucidum or the reishi mushroom reportedly antagonized the suppression on lymphocytes induced by culture supernatants of B16F10 melanoma cells, suggesting their potential for cancer immunotherapy (Sun et al., 2011). Anti-tumor effects against ovarian cancer cells and enhancement of their sensitivity to cisplatin have also been reported for this mushroom (Zhao et al., 2011). Pre-administration of spores of the mushroom led to alleviation of oxidative stress and mitochondrial dysfunction in rat hippocampus injected intracerebroventricularly with streptozotocin, protected neurons from apoptosis, and improved cognitive dysfunction in Sprague-Dawley rats, suggesting that the mushroom may prove useful in prevention of Alzheimer's disease (Zhou et al., 2011). A lanostanoid triterpene, ganodermanontriol, has been isolated from this mushroom, which reportedly suppressed growth of colon cancer cells (Jedinak et al., 2011). A polysaccharide extract of the reishi mushroom demonstrated immunomodulatory and adjuvant activities, both in vivo as well as in vitro (Lai et al., 2010). Methyl chloride extract of the mushroom showed presence of a number of compounds with antinociceptive activities, including ganoderic acids A, B, G and H (Koyama et al., 1997).
The objective of the present study was to evaluate the potential of crude methanol extracts of Lentinula edodes and Ganoderma lucidum in alleviating pain in acetic acid-induced gastric pain writhings in Swiss albino mice.
Materials and methods
Mushrooms and Methanol Extraction:
Fresh Lentinula edodes and Ganoderma lucidum mushrooms were collected from the National Mushroom Development and Extension Centre in Savar, Dhaka district, Bangladesh in December 2010. The mushrooms were cut separately into small pieces and air-dried in the shade for 120 hours, grounded into a fine powder, and were extracted with methanol at a ratio of 1:8 (w/v) for Lentinula edodes and at a ratio of 1:10 (w/v) for Ganoderma lucidum. After 24 hrs, the mixture was filtered; filtrate was collected and evaporated to dryness. The initial dry weight of Lentinula edodes used for extraction was 100g; the final weight of the extract was 17.0g. The initial dry weight of Ganoderma lucidum used for extraction was 100g; the final weight of the extract was 3.47g.
Chemicals and Drugs
Glacial acetic acid was obtained from Sigma Chemicals, USA; aspirin was obtained from Square Pharmaceuticals Ltd., Bangladesh. All other chemicals were of analytical grade.
In the present study, Swiss albino mice (male), which weighed from 20-25g were used. The animals were obtained from International Centre for Diarrheal Disease Research, Bangladesh (ICDDR,B). All animals were kept under ambient temperature with 12h light followed by a 12h dark cycle. The animals were acclimatized for three days prior to actual experiments. The study was conducted following approval by the Institutional Animal Ethical Committee of University of Development Alternative, Dhaka, Bangladesh.
Acetic Acid-induced Writhing Method
Antinociceptive activity of methanol extract of Lentinula edodes and Ganoderma lucidum was examined using previously described procedures of Shanmugasundaram and Venkataraman (2005) with minor modifications. Pain was induced in mice in the writhing test through intraperitoneal administration of 1% acetic acid at a dose of 10 ml/kg body weight. Mice were separated into nine groups of six mice each. Group-I served as control and was administered vehicle (1% Tween 80 in water, 10 ml/kg body weight). The standard drug, aspirin was administered to Group-II mice at a dose of 400 mg/kg body weight. Groups-III to V received extract of Lentinula edodes, respectively at 50, 200, and 400 mg extract/kg body weight orally 30 min before acetic acid injection. Groups-VI to X received extract of Ganoderma lucidum, respectively at 50, 100, 200, and 400 mg extract/kg body weight orally 30 min before acetic acid injection. A period of 5 minutes was given to each animal to ensure bio-availability of acetic acid, following which period, the number of writhings was counted for 10 min.
Experimental values are expressed as mean [+ or -] SEM. Independent Sample t-test was carried out for statistical comparison. Statistical significance was considered to be indicated by a p value < 0.05 in all cases.
Results and discussion
The methanol extract of Lentinula edodes dose-dependently reduced the number of writhings in mice injected intraperitioneally with acetic acid, as compared to control animals. At extract doses of 50, 200, and 400 mg per kg body weight, the number of writhings were respectively, reduced by 33.7, 51.8, and 54.2%. The results were statistically significant at the two higher doses of the extract tested. In comparison, the standard antinociceptive drug, aspirin, when administered to mice at a dose of 400 mg per kg body weight, reduced the number of writhings by 49.4%. Thus the two higher doses of Lentinula edodes demonstrated a higher efficiency in relieving pain than aspirin, the latter when administered at a dose of 400 mg per kg body weight.
The methanol extract of Ganoderma lucidum also exhibited significant reductions in the number of writhings in acetic acid-injected mice at doses of 200 and 400 mg per kg body weight. At the two lower doses of 50 and 100 mg per kg body weight, the extract did not show any effects in relieving gastric pain. The highest inhibition of writhings was observed at a dose of 400 mg extract per kg body weight. At this extract dose, the number of writhings was reduced by 36.1%. Taken together, extracts of both Lentinula edodes and Ganoderma lucidum exhibited significant antinociceptive properties, and can be considered for further studies towards isolation of relevant pharmacologically active components. It may be noted that the methyl chloride extract of Ganoderma lucidum has already been reported for containing compounds with antinociceptive effects. The present study indicates that the methanol extract may also contain compound(s) with antinociceptive effects.
Analgesia can be central or peripheral, and both central and peripheral analgesia can be detected with the acetic acid-induced writhing test (Shanmugasundaram and Venkataraman, 2005). Prostaglandins are considered as promulgators of pain. Production of prostaglandins [mainly prostacyclines ([PGI.sub.2]) and prostaglandin- (PG-E)] has been shown to be responsible for excitation of Ad-nerve fibers, leading to the sensation of pain (Reynolds, 1982; Rang and Dale, 1993). As such, the antinociceptive activity exhibited by crude methanolic extracts of the two mushroom species may be due to these extract's ability to block synthesis of prostaglandins. This, in turn, may be mediated through inhibition of cyclooxygenase and/or lipooxygenase activities. It is to be noted that a similar mechanism has been proposed for antinociceptive activity of Ficus deltoidea aqueous extract in acetic acid-induced gastric pain model (Sulaiman et al., 2008).
Mushrooms are not considered a delicacy by the general people of Bangladesh. Because of their nutritional properties and effects on improvement of health as a result of consumption, the Government is trying to promote cultivation and consumption of edible mushroom species within the general population of Bangladesh. At the same time, numerous studies have reported on the existence of chemical components in various mushroom species, which can be of value both as functional food as well as therapeutics. The present study suggests that the two mushroom species evaluated, can be of importance in the discovery of novel compounds for relief of pain. Pain due to various factors is a universal phenomenon, and any new compound which can relieve pain in an efficient manner without causing unwelcome side-effects can prove to be of real benefit to human beings.
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Corresponding Author: Mohammed Rahmatullah, Faculty of Life Sciences University of Development Alternative House No. 78, Road No. 11A (new) Dhanmondi, Dhaka-1205 Bangladesh Tele +88-01715032621; Fax: +88-02-815739 E-mail: email@example.com
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(1) Fatema Toz Zohra, (1) Nusrat Harun Antara, (1) Shahnaz Rahman, (1) Aynal Haq Rana, (1) Farjana Akther Noor, (1) Himel Nahreen Khaleque, (1) Sherezad Sanam, (1) Farhana Islam, (1) Alok Kumar Paul, (2) Nirod Chandra Sarker, (1) Mohammed Rahmatullah
(1) Faculty of Life Sciences, University of Development Alternative, Dhanmondi, Dhaka-1205, Bangladesh. (2) National Mushroom Development and Extension Centre, Savar, Dhaka, Bangladesh.
Table 1: Antinociceptive effect of crude methanol extract of Lentinula edodes and Ganoderma lucidum mushroom species in the acetic acid-induced gastric pain model mice. Dose (mg/kg Mean number Treatment body weight) of writhings % inhibition Control 10 ml 8.33 [+ or -] 0.56 -- Aspirin 400 mg 4.17 [+ or -] 1.38 49.4 * Lentinula edodes 50 mg 5.50 [+ or -] 1.73 33.7 Lentinula edodes 200 mg 4.00 [+ or -] 1.15 51.8 * Lentinula edodes 400 mg 3.83 [+ or -] 0.87 54.2 * Ganoderma lucidum 50 mg 8.67 [+ or -] 0.84 -- Ganoderma lucidum 100 mg 8.50 [+ or -] 0.56 -- Ganoderma lucidum 200 mg 5.50 [+ or -] 1.15 33.7 * Ganoderma lucidum 400 mg 5.33 [+ or -] 1.31 36.1 * All administrations (aspirin and extract) were made orally. Values represented as mean [+ or -] SEM, (n=6); * P < 0.05; significant compared to control.
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|Title Annotation:||Original Article|
|Author:||Zohra, Fatema Toz; Antara, Nusrat Harun; Rahman, Shahnaz; Rana, Aynal Haq; Noor, Farjana Akther; Kha|
|Publication:||Advances in Natural and Applied Sciences|
|Date:||Apr 1, 2011|
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