EVALUATION OF ACUTE AND SUB-ACUTE TOXICITIES OF GANODERMA LUCIDUM
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1 EVALUATION OF ACUTE AND SUB-ACUTE TOXICITIES OF GANODERMA LUCIDUM Sheena.N Studies on the thekapeutic potential of Ganoderma lucidum P. Karst Reishi, occurring in Kerala Thesis. Amala Cancer Research Centre, University of Calicut, 2005
2 CHAPTER 7 I1 EVALUATION OF ACUTE AND SUB-ACUTE TOXICITIES i: OF GANODERMA L UCIDUM
3 7,1, INTRODUCTION Medicinal mushrooms and mushroom metabolites have notable place in the folklore throughout the world. They are traditionally used in China and Japan for medicinal and tonic properties (Wasser et al., 2000., Mizuno, 2000., Begell and Wasser, 2001., Cimerman, 1999). Species of genus Ganoderma P. Karst. is used in folk medicine for thousands of years in the folklore in China and persistent use of medicinal mushrooms among people of China could be considered a good evidence of their efficacy. Although there are several reports regarding the frequent use of mushrooms in the treatment of a number of disease conditions. Experimental studies, which validate the toxicity studies of mushrooms are rare. Mushrooms are promising resources of physiologically functional food and materials for the development of medicines, pharmaceutical products such as new drugs, dietary supplements and healthy beverages, cosmetic products, among others (Lomderg, et al., 2001). In the last decade medicinal mushrooms were intensively investigated for medicinal effects in in vivo and in vitro model systems, and pharmacologically active substances were identified. Ganoderma lucidum (W. Curt.: Fr.) Karst. most commonly known as reishi, is a wood rotting fungus generally found growing on trees and stumps is prescribed by practitioners of Traditional Chinese Medicine for the treatment of neurasthenia, debility from prolonged illness, insomnia, anorexia, dizziness, chronic hepatitis, hypercholesterolemia, coronary heart disease, hypertension, altitude sickness,
4 fatigue, carcinoma, and bronchial cough (Ma et al., 2002). Investigations were carried out on the acute and sub acute toxicity of aqueous methanol extract of G. lucidum occurring in Kerala. The findings are reported in this chapter. 7,2, MATERIALS AND METHODS PREPARATION OF THE EXTRACT Aqueous methanol extract of G. lucidum was prepared as described in the section ANIMALS Toxicity studies : Male Swiss albino mice weighing 25 k 2 g ACUTE TOXICITY STUDY Mice were divided into three groups of six animals each. The drug was administered orally as single dose as follows (Walum, 1998). Group 1 Group 11 Group I11 Normal Aqueous methanol extract 500 mglkg bodyweight Aqueous methanol extract 2500 mglkg bodyweight The animals were observed for toxic symptoms and mortality for 72 h SUB-ACUTE TOXICITY STUDY The mice were divided into three groups of six animals each. The drug was administered to animals once daily for 30 days (Parchment, 1998). Group 1 Group I1 Normal Aqueous methanol extract 500 mglkg bodyweight
5 Group I11 Aqueous methanol extract 1500 mglkg bodyweight The animals were observed for toxic symptoms and mortality. Group 1 treated with distilled water was kept as normal. One day after the last dose of extract administration animals were sacrificed. The blood was collected from the heart. Hematological parameters for total erythrocytes count (section ), total leukocytes count (section ) and haemoglobin (section ) were determined. Serum was used for the determination of liver function test, transaminases, SGOT (section ) and SGPT (section ), alkaline phosphatase (section ), and renal function tests such as urea (section ) and creatinine (section ) RESULTS ASSAY FOR ACUTE TOXICITY STUDY The animals administered with aqueous methanol extract of G.lucidum did not produce any external symptoms of toxicity or mortality up to the dose of 2500 mglkg bodyweight orally ASSAY FOR SUB-ACUTE TOXICITY STUDY In sub acute toxicity studies, treatment of extract also did not produce any statistically significant change in the hematological or biochemical parameters when compared to the normal group of animals. However, treatment of aqueous methanolic extract (1500 mglkg bodyweight) for 30 days produces a slight increase
6 in the WBC, RBC and Hb count (Table 7.1). Similarly the administration of extract did not produce any significant changes in the liver function tests (SGOT, SGPT) and renal function tests (urea and creatinine). The SGOT and SGPT activities in the normal and the extract treated (1500 mgkg body wt) were (35. 2 h 1.8 IUI1 and 37.5 * 2.7 IU11) and (100.2 * 6 IUIl and * 3.2 IU11) (Table 7.2). The ALP level in the normal and the extract treated (1500 mgkg body wt) was 70.7 h 16.4 IU11 and 74.5 * 6.7 IU11 (Table 7.2). The normal animals showed serum urea level 69.4 * 8.7 mgldl and creatinine level 0.43 * 0.02 mgldl, while the extract (1500 mgkg body wt) treated group showed 78.0 * 8.1 mgldl and 0.44 * 0.01 mgldl (Table 7.3).
7 Table 7.1. Effect of aqueous methanol extract of G. lucidum on total WBC, RBC counts and haemoglobin concentration. Groups Treatments WBC RBC (Cells1 p1) (Cells1 p])) (gfdl) Normal * * k k * 0.8 G. lucidum Extract * * * 0.54 Values are mean S. D. n= 6 animals. Treatments are not significantly different
8 Table 7.2. Effect of aqueous methanol extract of G. lucidum on serum SGOT, SGPT and ALP levels. Groups Treatments SGOT SGPT ALP (mglkg) (IUII) (IUII) (IUII) Normal k k k k k lk 14.3 G. lucidum Extract * * S 6.7 Values are mean S. D. n= 6 animals. Treatments are not significantly different
9 Table 7.3. Effect of aqueous methanol extract of G. lucidum on serum urea and creatinine concentration Groups Treatments Urea Creatinine (mglkg) (mgldl) (mg/dl) Normal f k * * 0.03 G. lucidum Extract * k 0.01 Values are mean k S. D. n= 6 animals. Treatments are not significantly different
10 Results of the study reveal that aqueous methanolic extract of G. lucidum showed no signs of toxicity. The extract administered up to 1500 mglkg body wt. was not lethal to animals. Similarly the administration of the extract is not found to produce any hematological toxicity. Liver function and renal function tests show no significant change in the serum SGOT, SGPT, ALP and also serum urea and creatinine levels in the extract treated group from the normal group support the safety of the extract for therapeutic use. Many pharmaceutical substances with potent and unique valuable properties have been isolated recently from mushrooms and distributed worldwide (Wasser et al., 2000). Biologically active polysaccharides are the best-known mushroom derived substances with antitumor properties (Reshetnikov et al., 2001). G. lucidum is most commonly used in traditional Chinese medicine for the treatment of various diseases (Ma et al., 2002). But a clear picture of its toxicokinetics is still obscure and the present study is an attempt to validate the safety of G. lucidum extract. The current experimental findings reveal non- toxic nature of G. lucidum occurring in Kerala and its potential therapeutic use.
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