Preclinical toxicological evaluations of the sclerotium of Lignosus rhinocerus (Cooke), the Tiger Milk mushroom
Graphicak abstract
Introduction
Lignosus rhinocerus (Synonym Lignosus rhinocerotis (Cooke) Ryvarden), commonly known as Tiger Milk mushroom, belongs to the Polyporaceae family. Its geographical distribution is only in the tropical rainforest in the region of South China, Thailand, Malaysia, Indonesia, Philippines and Papua New Guinea (Tan et al., 2012). In Malaysia, it is also known as ‘Cendawan susu rimau’ and is the most popular medicinal mushroom used by the indigenous communities of Peninsular Malaysia to relieve fever, cough, asthma, cancer, food poisoning and as a general tonic (Lee et al., 2009). In China, the sclerotium of the mushroom is an expensive traditional medicine used for treatment of liver cancer, chronic hepatitis and gastric ulcers (Wong and Cheung, 2008). Phylogenetic analysis indicated that the mushroom is closely related to Ganoderma lucidum and Trametes versicolor, the two most popular medicinal mushrooms used in Asia (Tan et al., 2010).
The sclerotium of Lignosus rhinocerus has been demonstrated to exhibit anti-proliferative activity. Lai et al. (2008) demonstrated that sclerotial polysaccharides from Polyporus rhinocerus (synonym of Lignosus rhinocerus) exhibited antiproliferative effects on several kinds of leukemic cell lines. Wong et al. (2009) reported that the hot water extract of Polyporus rhinocerus exhibited immunomodulatory effect by stimulating human innate immune cells. Our studies demonstrated that the cold water extract of Lignosus rhinocerus sclerotium exhibited direct cytotoxicity on human breast carcinoma (MCF-7) and human lung carcinoma (A549) cell lines (Lee et al., 2012a). It has also been demonstrated that the sclerotial extracts of Lignosus rhinocerus exhibited anti-acute inflammatory activity in an animal model study (Lee et al., 2012b). Gao et al. (2009) showed that the non-digestible carbohydrates may function as novel prebiotics. Recently, aqueous sclerotial extract of Lignosus rhinocerus was reported to contain neuroactive compounds that stimulated neurite outgrowth in PC-12 cell line (Eik et al., 2012).
In view of its wide ethno-botanical usages as well as proven in vitro anti-proliferative and anti-inflammatory activities, Lignosus rhinocerus sclerotium may be used as a health supplement. It is therefore necessary to carry out in depth safety evaluation of the sclerotium. Earlier studies to evaluate the subacute toxicity of the sclerotial powder of Lignosus rhinocerus cultivar (termed TM02) demonstrated its no-observed-adverse-effect level (NOAEL) was higher than 1000 mg/kg in a 28 days animal studies using rats. In the present study, we carried out a 180 day chronic toxicity study of the sclerotial powder, as well as evaluating its possible anti-fertility and teratogenic effects and genetoxicity.
Section snippets
Preparation of Lignosus rhinocerus sclerotial powder
Sclerotial powder of the Lignosus rhinocerus cultivar TM02 was provided by Ligno Biotech Sdn. Bhd. (Selangor, Malaysia). The fungus was identified by internal transcribed spacer (ITS) regions of the ribosomal DNA (Tan et al., 2010). The sclerotial powder was freeze-dried and milled into powder using 0.2 mm sieve. The powder is light brown, dry fluffy powder with milk like taste.
Animals
Sprague Dawley (SD) rats aged 5 weeks old (male and female) were supplied by Chenur Supplier (Selangor, Malaysia). The
Body weight and general clinical observations
Oral administration of the sclerotial powder of Lignosus rhinocerus (cultivar TM02) at all the doses tested did not produce any treatment related abnormality in the rats, and there were no death observed. Fig. 1, Fig. 2 show the body weight of male and female rats treated with various doses of the sclerotial powder, and that of the control group for 180 days (26 weeks). The net body weight gain of the all the treated groups were not significantly different from the control animals (data not
Discussion
Changes in body weight have been used as an indicator of adverse effects of drugs and chemicals (Hilaly et al., 2004, Mukinda and Eagles, 2010). In the present studies, the similar growth pattern as shown by body weight (Fig. 1, Fig. 2) and body weight gain (data was not shown) for the treated and the control groups indicated that oral administration of sclerotial powder of Lignosus rhinocerus (TM02) at a daily dose of up to 1000 mg/kg and for 180 days had no adverse effect on the growth of the
Conclusions
Our results showed that there are no treatment-related chronic toxicity in rats of both sexes following the long term (180-days) oral administration of 250, 500 and 1000 mg/kg of Lignosus rhinocerus (TM02) sclerotial powder, as shown by the clinical observations, body weight gain, hematological analysis, clinical biochemistry, urinalysis, absolute organ weight as well as relative organ weight, and histological examinations of the organs. Thus, the no-observed-adverse-effect level (NOAEL) dose of
Acknowledgements
This research is supported by MOHE-UM HIR E20040-20001 and PV054-2011B from University of Malaya. The authors would like to express gratitude to staff from Laboratory Animal Center, Faculty of Medicine, University of Malaya for their assistance.
References (22)
- et al.
Acute and chronic toxicological studies of Ajuga iva in experimental animals
Journal of Ethnopharmacology
(2004) - et al.
Utilization of macrofungi by some indigenous communities for food and medicine in Peninsular Malaysia
Forest Ecology and Management
(2009) - et al.
Evaluation of the sub-acute toxicity of the sclerotium of Lignosus rhinocerus (Cooke), the Tiger Milk mushroom
Journal of Ethnopharmacology
(2011) - et al.
Acute and sub-chronic oral toxicity profiles of the aqueous extract of Polygala fruticosa in female mice and rats
Journal of Ethnopharmacology
(2010) - et al.
Pharmacologic and toxicologic effects of two Maytenus species in laboratory animals
Journal of Ethnopharmacology
(1991) - et al.
Concordance of the toxicity of pharmaceuticals in humans and in animals
Regulatory Toxicology and Pharmacology.
(2000) - Commission Regulation (EC), 2008. Mutagenicity-Reverse Mutation Test Using Bacteria. No. 440/2008 (B.13/14.). Adopted...
- et al.
Lignosus rhinocerus (Cooke) Ryvarden: a medicinal mushroom that stimulates neurite outgrowth in PC-12 cells
Evidence-Based Complementary and Alternative Medicine
(2012) - et al.
Nondigestible carbohydrates isolated from medicinal mushroom sclerotia as novel prebiotics
International Journal of Medicinal Mushrooms.
(2009) - et al.
Antiproliferative effects of sclerotial polysaccharides from Polyporus rhinocerus Cooke (Aphyllophoromycetideae) on different kinds of leukemic cells
International Journal of Medicinal Mushrooms
(2008)
The antiproliferative activity of sclerotia of Lignosus rhinocerus (Tiger Milk Mushroom)
Evidence-Based Complementary and Alternative Medicine
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