Ganoderma lucidum total triterpenes attenuate DLA induced ascites and EAC induced solid tumours in Swiss albino mice.

G. lucidum total triterpenes were assessed for its apoptosis-inducing and anti-tumour activities. The ability of the total triterpenes to induce apoptosis was evaluated in Dalton's lymphoma ascites (DLA) and Ehrlich's ascites carcinoma (EAC) cell lines. Total triterpenes were found to be highly cytotoxic to DLA and EAC cell lines with IC50 values 5 ± 0.32 and 7.9 ± 0.2 µg/ml respectively. Total triterpenes induced apoptosis in both cell lines which is evident from the DNA fragmentation assay. Anti-tumour activity was accessed using DLA induced solid and EAC induced ascites tumour models in Swiss albino mice. Administration of 10, 50 and 100 mg/kg b. wt. total triterpenes showed 11.86, 27.27 and 40.57% increase in life span of animals in ascites tumour model. Treatment with 10, 50 and 100 mg/kg b. wt. total triterpenes exhibited 76.86, 85.01 and 91.03% inhibition in tumour volume and 67.96, 72.38 and 77.90% inhibition in tumour weight respectively in the solid tumour model. The study reveals the significant dose-dependent anti-tumour activity of total triterpenes in both models. Total triterpenes were more active against the solid tumour than the ascites tumour. The anti-oxidant potential and ability to induce cell-specific apoptosis could be contributing to its anti-tumour activities.

Ganoderma lucidum total triterpenes prevent γ-radiation induced oxidative stress in Swiss albino mice in vivo.

OBJECTIVES: The in vivo radio-protective effect of total triterpenes isolated from Ganoderma lucidum (Fr.) P. Karst was evaluated using Swiss albino mice, by pre-treatment with total triterpenes for 14 days, followed by a whole body exposure to γ-radiation. METHODS: The activities of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), and the level of reduced glutathione (GSH) were analysed in liver and brain homogenates. The extent of lipid and protein peroxidation was also estimated in liver and brain homogenates after irradiation. Protection of radiation-induced DNA strand breaks in peripheral blood lymphocytes and bone marrow cells was assessed using the comet assay. RESULTS: Total triterpenes were highly effective in reducing the levels of lipid peroxidation and protein oxidation to near normal values in both liver and brain tissues. Total triterpenes, when administered in vivo, were also found to be successful in restoring the antioxidant enzyme activities and GSH level in liver and brain of irradiated mice. Administration of total triterpenes, prior to radiation exposure, significantly decreased the DNA strand breaks. DISCUSSION: The results of the present study thus revealed the potential therapeutic use of Ganoderma total triterpenes as an adjuvant in radiation therapy.

Ganoderma lucidum ameliorate mitochondrial damage in isoproterenol-induced myocardial infarction in rats by enhancing the activities of TCA cycle enzymes and respiratory chain complexes.

BACKGROUND: Decreased mitochondrial function has been suggested to be one of the important pathological events in isoproterenol (ISO)-induced cardiotoxicity. In this communication, we have evaluated the protective effect of Ganoderma lucidum against ISO induced cardiac toxicity and mitochondrial dysfunction. METHODS: Cardiac toxicity was assessed by determining the activities of creatine kinase (CK) and lactate dehydrogenases (LDH) after subcutaneous injection of ISO (85 mg/kg) at an interval of 24h for 2 days. The animals were sacrificed 24h after last ISO administration. G. lucidum (100 and 250 mg/kg, p.o.) was given to the rats once daily for 15 days prior to the ISO challenge. Similarly, α-Tocopherol (100mg/kg, p.o) was kept as the standard. To assess the extent of cardiac mitochondrial damage, the activities of Krebs cycle dehydrogenases and mitochondrial complexes I, II, III, and IV as well as the level of ROS and mitochondrial membrane potential (ΔΨmt) were evaluated. RESULTS: Administration of G. lucidum and α-tocopherol significantly protected the elevated activities of CK and LDH. Further, the activities of mitochondrial enzymes and the level of ΔΨmt were significantly enhanced and the level of ROS was significantly declined in the G. lucidum and α-tocopherol treatments. CONCLUSION: The present study concluded that the cardiac mitochondrial enzymes are markedly declined by the ISO challenge and the administration G. lucidum and α-Tocopherol significantly protected mitochondria by preventing the decline of antioxidant status and ΔΨmt or by directly scavenging the free radicals.

Hepatoprotective activity of cultured mycelium of Morel mushroom, Morchella esculenta.

The hepatoprotective activity of cultured mycelium of morel mushroom Morchella esculenta against CCl(4) and ethanol induced chronic hepatotoxicity was investigated. Hepatotoxicity was induced by challenging the animals with CCl(4) (1:5, v/v, 3.75 ml/kg body weight, i.p., 30 doses) and ethanol (36%, v/v, 6 ml/animal, p.o., 35 doses) and the extract was administered at two concentrations (250 and 500 mg/kg body weight). Hepatoprotection was evaluated by determining the activities of liver function marker enzymes and antioxidant status of liver and also by histopathological observations of liver tissue. Administration of both ethanol and CCl(4) elevated the levels of liver function enzymes, GOT, GPT and ALP in serum drastically. The treatment with the extract decreased the elevated serum GOT, GPT and ALP activities in a dose dependent manner. The extract also restored the depleted levels of antioxidants in liver consequent to CCl(4) and ethanol challenge. The results indicated that aqueous-ethanolic extract of M. esculenta mycelium possessed significant hepatoprotective activity. The conclusion is also supported by the biochemical determinations and histopathological observations. The findings thus suggest the potential therapeutic use of morel mushroom mycelium as a novel hepatoprotective agent.

Antioxidant activity and toxicity profile of total triterpenes isolated from Ganoderma lucidum (Fr.) P. Karst occurring in South India.

The total triterpene fraction isolated from Ganoderma lucidum, a highly nutritional and popular medicinal mushroom occurring in South India, was evaluated for its antioxidant activity in vitro and in vivo. Total triterpenes successfully scavenged DPPH(+), ABTS(+) and superoxide radicals, showed significant ferric reducing activity and was highly effective in reducing the in vitro lipid peroxidation. Activities of the antioxidant enzymes in blood and tissue were increased by the administration of total triterpenes to Swiss albino mice in vivo. The ability of total triterpenes to scavenge the free radicals and to enhance body's antioxidant defence systems indicates its potential use as an antioxidant. An attempt was also done to gauge the toxicity of total triterpenes using acute and sub acute study models in Swiss albino mice. The results showed that Ganoderma triterpenes did not possess significant toxicity. The findings thus reveal the possible therapeutic use of Ganoderma triterpenes.

Evaluation of free radical scavenging activity of morel mushroom, Morchella esculenta mycelia: a potential source of therapeutically useful antioxidants.

Cellular damage caused by reactive oxygen species (ROS) has been implicated in several diseases and antioxidants are known to protect the body from this damage. Antioxidants thus, have gained significant importance in human health. The search for effective, non-toxic natural compounds with antioxidant activity has intensified in recent years. Mycelia of a number mushrooms have recently been successfully used for the development of novel pharmaceutical products. We examined the aqueous-ethanol extract of cultured mycelia of the morel mushroom, Morchella esculenta (L.) Pers. (Morchellaceae) for its ability to scavenge super oxide, hydroxyl, nitric oxide, 2,2'-diphenyl-1-picrylhydrazyl (DPPH), and 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radicals as well as for inhibition of lipid peroxidation. The extract efficiently scavenged all these radicals and also inhibited lipid peroxidation. Ferric reducing antioxidant power (FRAP) assay indicated the hydrogen donating capacity of the extract. The pulse radiolysis studies using ABTS and carbonate radical (CO(3)(*-)) showed that the extract significantly carried out the decay of these radicals in a concentration-dependent manner. In conclusion, the investigation showed that the morel mushroom mycelium is an excellent source of antioxidants which are capable of imparting protection at different levels. The findings suggest the potential therapeutic use of morel mushroom, M. esculenta mycelia as an efficient antioxidant.

Therapeutic potential of Ganoderma lucidum (Fr.) P. Karst. against the declined antioxidant status in the mitochondria of post-mitotic tissues of aged mice.

BACKGROUND & AIMS: Post-mitotic cells such as brain and heart cells are particularly vulnerable to oxidative damages during ageing. In this study, we evaluated the effect of Ganoderma lucidum on the antioxidant status in the mitochondria of heart and brain of aged mice. METHODS: The effect was evaluated by estimating the activities of manganese-superoxide dismutase (Mn SOD), glutathione peroxidase (GPx), glutathione-S-transferase (GST) and catalase (CAT) as well as levels of reduced glutathione (GSH), lipid peroxidation, advanced oxidation protein products (AOPP) and reactive oxygen species (ROS) in the heart and brain mitochondria of aged mice after oral administration of ethanolic extract of G. lucidum (50 and 250mg/kg), once daily for 15 days. The effect was compared with that of aged and young control animals. dl-alpha-lipoic acid (100mg/kg) was taken as the positive control. RESULTS: Administration of G. lucidum extract significantly (p<0.05) elevated the levels of GSH as well as activities of Mn SOD, GPx, and GST and decreased significantly (p<0.05) the levels of lipid peroxidation, AOPP and ROS. CONCLUSION: G. lucidum administration could improve the age-related decline of antioxidant status which was partly ascribed to free radical scavenging activity.

Palladium alpha-lipoic acid complex formulation enhances activities of Krebs cycle dehydrogenases and respiratory complexes I-IV in the heart of aged rats.

Age-related decline in the capacity to withstand stress, such as ischemia and reperfusion, results in congestive heart failure. Though the mechanisms underlying cardiac decay are not clear, age dependent somatic damages to mitochondrial DNA (mtDNA), loss of mitochondrial function, and a resultant increase in oxidative stress in heart muscle cells may be responsible for the increased risk for cardiovascular diseases. The effect of a safe nutritional supplement, POLY-MVA, containing the active ingredient palladium alpha-lipoic acid complex, was evaluated on the activities of the Krebs cycle enzymes such as isocitrate dehydrogenase, alpha-ketoglutarate dehydrogenase, succinate dehydrogenase, and malate dehydrogenase as well as mitochondrial complexes I, II, III, and IV in heart mitochondria of aged male albino rats of Wistar strain. Administration of 0.05 ml/kg of POLY-MVA (which is equivalent to 0.38 mg complexed alpha-lipoic acid/kg, p.o), once daily for 30 days, was significantly (p<0.05) effective to enhance the Krebs cycle dehydrogenases, and mitochondrial electron transport chain complexes. The unique electronic and redox properties of palladium alpha-lipoic acid complex appear to be a key to this physiological effectiveness. The results strongly suggest that this formulation might be effective to protect the aging associated risk of cardiovascular and neurodegenerative diseases.

Ganoderma lucidum (Fr.) P. Karst enhances activities of heart mitochondrial enzymes and respiratory chain complexes in the aged rat.

Aging is associated with increased oxidative damage at multiple cellular levels, decline in cellular energy production and enhanced free radical status. The effect of the medicinal mushroom, Ganoderma lucidum on the activities of tricarboxylic acid (Krebs) cycle enzymes and mitochondrial complexes I-IV of the electron transport chain in aged rats were investigated. The activity of Krebs cycle enzymes, isocitrate dehydrogenase, alpha-ketoglutarate dehydrogenase, succinate dehydrogenase, and malate dehydrogenase as well as mitochondrial complexes I, II, III, and IV were determined in heart of aged male Wistar rats orally administrated with 70% ethanolic extract (50 and 250 mg/kg) of G. lucidum. DL-alpha-lipoic acid (100 mg/kg) was taken as the positive control. Administration of the G. lucidum, once daily for 15 days, was significantly (P < 0.05) effective to enhance the Krebs cycle dehydrogenases, and mitochondrial electron transport chain complex IV activities in aged rats. The profound activity of the extract can be correlated to the significant antioxidant property of G. lucidum. The results of the study revealed that G. lucidum is effective to ameliorate the age associated decline of cellular energy status.

Effect of Ganoderma lucidum on the activities of mitochondrial dehydrogenases and complex I and II of electron transport chain in the brain of aged rats.

Dysfunction of the mitochondrial respiratory chain, being direct intracellular source of reactive oxygen species (ROS), is important in the pathogenesis of number of ageing associated human disorders. Effect of ethanol extract of Ganoderma lucidum on the activities of mitochondrial dehydrogenases; complex I and II of electron transport chain have been evaluated in the aged rat brain. Aged male Wistar rats were administered with ethanol extract of G. lucidum (50 and 250mg/kg, p.o) once daily for 15 days. Similarly DL-alpha-lipoic acid (100mg/kg, p.o) administered group was kept as the reference standard. Young and aged rats administered with water were kept as young and aged control, respectively. The effect of treatment was assessed by estimating the activities of succinate dehydrogenase (SDH), malate dehydrogenase (MDH), alpha-ketoglutarate dehydrogenase (alpha-KGDH), pyruvate dehydrogenase (PDH), complex I and II in the mitochondria of rat brain. Results of the study demonstrated that the extract of G. lucidum (50 and 250mg/kg) significantly (p<0.01) enhanced the activities of PDH, alpha-KGDH, SDH, complex I and II when compared to that of the aged control animals. The level of the lipid peroxidation was significantly lowered (p<0.01) in the G. lucidum treated group with respect to that of aged control. However, we could not find any statistically significant difference between the activities of enzymes in groups treated with 50 and 250mg/kg of G. lucidum. The activity exhibited by the extract of G. lucidum in the present study can be partially correlated to its antioxidant activity. The results of the study concluded that the extract of G. lucidum may effective to improve the function of mitochondria in aged rat brain, suggest its possible therapeutic application against ageing associated neurodegenerative diseases.

Antimutagenic activity of methanolic extract of Ganoderma lucidum and its effect on hepatic damage caused by benzo[a]pyrene.

The antimutagenic activity of the methanolic extract of the fruiting bodies of Ganoderma lucidum (Fr.) P. Krast. occurring in South India was investigated. The activity was assayed by Ames Salmonella mutagenicity test using histidine mutants of Salmonella typhimurium tester strains, TA98, TA100 and TA102. The methanolic extract of the mushroom significantly inhibited (P<0.001) the in vitro sodium azide (NaN(3)), N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and 4-nitro-o-phenylenediamine (NPD), and benzo[a]pyrene (B[a]P) induced his(+) revertants in a dose dependent manner. In vivo antimutagenic activity of extract was also assayed by determining the mutagenicity of the urine of rats administrated with B[a]P as a mutagen. The prior administration of extract markedly inhibited mutagenicity induced by B[a]P. The results indicated that the methanolic extract of Ganoderma lucidum occurring in South India possessed significant antimutagenic activity. The effect of B[a]P on hepatic enzymes, such as serum glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT) and alkaline phosphtase (ALP), were also evaluated. The extract prevented the increase of SGOT, SGPT, and ALP activities consequent to B[a]P challenge, and enhanced the levels of reduced glutathione (GSH) and activities of glutathione peroxidase (GPx), glutathione-S-transferase (GST), superoxide dismutase (SOD), and catalase (CAT). The extract also profoundly inhibited lipid peroxidation induced by B[a]P. The results revealed that Ganoderma lucidum extract restored antioxidant defense and prevented hepatic damage consequent to the challenge by B[a]P.

Methanol extract of the oyster mushroom, Pleurotus florida, inhibits inflammation and platelet aggregation.

The antiinflammatory and platelet aggregation inhibiting activities of the methanol extract of Pleurotus florida Eger, an edible and commercially grown mushroom, were investigated. The extract showed significant activity in ameliorating acute inflammation induced by carrageenan and chronic inflammation by formalin at 500 and 1000 mg/kg body weight. The effect was comparable to the standard reference drug, diclofenac. The extract also showed significant platelet aggregation inhibiting activity of washed human platelets. Adenosine 5'-diphosphate (ADP) induced platelet aggregation was inhibited by 88% to 95% at a concentration of 500 microg/mL of the extract after a preincubation time of 5, 10 and 20 min. The marked antiinflammatory and platelet aggregation inhibiting properties of this mushroom suggest its potential therapeutic use against vascular disorders.

Potential risk of acute hepatotoxicity of kodo poisoning due to exposure to cyclopiazonic acid.

Kodo millet (Paspalum scrobiculatum L.) is a staple food of some sections of people of North India. Consumption of Kodo millet is often found to cause intoxication and poisoning. The grains are frequently infested with Aspergillus tamarii Kita, which produced substantial amount of a mycotoxin, cyclopiazonic acid (CPA). Investigations were carried out to evaluate the hepatotoxic/preneoplastic changes in rat liver following single and multiple dose administration of CPA. Results showed a marked increase in the activity of glutamate pyruvate transaminase (GPT) and glutamate oxaloacetate transaminase (GOT) following CPA exposures, suggesting acute hepatotoxicity. Significant increase was also observed in gamma glutamyl transpeptidase (GGT) activity following CPA exposures, indicating preneoplastic changes in the liver. The results reveal that Kodo poisoning might cause acute hepatotoxicity in men and animals. The findings thus suggest that the consumption of contaminated Kodo millet is a serious health hazard due to exposure to CPA produced by Aspergillus tamarii associated with the millet.

Cytotoxic and antitumor activities of a polypore macrofungus, Phellinus rimosus (Berk) Pilat.

Cytotoxic and antitumor activities of ethyl acetate, methanol and aqueous extracts of a wood inhabiting polypore macrofungus, Phellinus rimosus (Berk) Pilat. were studied. Ethyl acetate and methanol extracts showed in vitro cytotoxic activity against Dalton's lymphoma ascites (DLA) and Ehrlich's ascites carcinoma (EAC) cell lines. The aqueous extract did not exhibit cytotoxicity against the tested cell lines. All the three extracts were highly effective in inhibiting growth of solid tumor induced by DLA cell line in mice. However, the antitumor activity of ethyl acetate extract was higher than that of methanol and aqueous extracts. The ethyl acetate extract was also effective in preventing the EAC induced ascites tumor development in mice. The antitumor activity of all the three extracts against solid tumor at a dose of 50 mg/kg (p.o.) was comparable to the clinically used standard reference drug, cisplatin (4 mg/kg, i.p.). Pre-treatment of the extracts was also effective in inhibiting the tumor growth induced by DLA cell lines. The experimental results revealed that ethyl acetate extract of P. rimosus possessed significant antitumor activity. The findings thus suggest the potential use of this mushroom as antitumor agent.

Antiperoxidative, anti-inflammatory, and antimutagenic activities of ethanol extract of the mycelium of Ganoderma lucidum occurring in South India.

Free radical mediated genetic instability is widely thought to be a major etiological factor for initiation of carcinogenesis. Mushrooms represent a largely untapped source of powerful new pharmaceutical products. In the present study, we examined the antiperoxidative, anti-inflammatory, and antimutagenic activities of the ethanol extract of the mycelium of a medicinal mushroom, Ganoderma lucidum, occurring in south India. Antiperoxidative activity was evaluated using Fe(2+)-ascorbate-induced lipid peroxidation in rat liver homogenate and a phorbol ester (croton oil)-induced lipid peroxidation in mouse skin. Antiinflammatory activity was evaluated against carrageenan-induced acute and formalin-induced chronic inflammatory paw edema in mouse and phorbol ester-induced mouse skin inflammation. Antimutagenic activity was determined by the Ames mutagenicity assay using histidine mutant of Salmonella typhimurium strains TA 98, TA100, and TA102. Sodium azide (NaN(3)), N-methyl-N-nitro-N-nitrosoguanidine (MNNG), 4-nitro-o-phenylenediamine (NPD), and benzo[a]pyrene (B[a]P) were used as the mutagens. The extract showed significant inhibition of Fe(2+)-induced peroxidation of lipid in rat liver (IC(50) 510 +/- 22 microg/ml) and 37% inhibition of croton oil-induced peroxidation on the mouse skin at 20 mg/0.1 ml/skin. Carrageenan-induced acute and formalin-induced chronic inflammatory edema were inhibited by 56 and 60%, respectively, by the extract at 1,000 mg/kg body wt (i.p). The extract at a concentration of 5 mg/plate showed inhibition of mutagenicity elicited by direct acting mutagens, NaN(3) (55.5 and 75.7%) and MNNG (50.0 and 57.5%) for S. typhymurium strains TA100 and TA102, respectively. The extract at the same concentration also inhibited mutagenicity elicited by NPD (52.4 and 64.2%) and B[a]P (60.7 and 59.6%) for TA98 and TA100 strains, respectively. The B[a]P was activated in the presence of rat liver microsomal (S9) fraction. The results of our study revealed that ethanol extract of Ganoderma lucidum mycelium possessed significant antiperoxidative, antiinflammatory, and antimutagenic activities. The findings suggest a medicinal use for the ethanol extract of the mycelium of G. lucidum occurring in South India.

Amelioration of cisplatin induced nephrotoxicity in mice by ethyl acetate extract of a polypore fungus, Phellinus rimosus.

Pretreatment of ethyl acetate extract of Phellinus rimosus (Berk) Pilat protected cisplatin induced nephrotoxicity in mice. Single dose of the extract (25 and 50 mg/kg body weight) decreased cisplatin induced serum creatinine and urea levels. The cisplatin depleted renal antioxidant defence system, such as activity of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and concentration of reduced glutathione (GSH), were restored by the treatment of the extract. The enhanced renal antioxidant defence system prevented cisplatin induced tissue lipid peroxidation. The experimental results suggest that ethyl acetate extract of P. rimosus protected cisplatin induced nephrotoxicity, possibly by enhancing renal antioxidant status. The extract did not interfere with the antitumor efficacy of cisplatin.

Antioxidant and antihepatotoxic activities of Phellinus rimosus (Berk) Pilat.

The antioxidant and antihepatotoxic activities of a wood inhabiting macrofungus, Phellinus rimosus were studied. The superoxide anion scavenging, Fe(2+)-ascorbate induced lipid peroxidation inhibiting, hydroxyl radical scavenging and nitric oxide scavenging activities of the ethyl acetate extract were determined. The results indicated that ethyl acetate extract of P. rimosus exhibited significant in vitro antioxidant activity. The ethyl acetate extract of P. rimosus also showed potent antihepatotoxic activity against carbontetrachloride-induced acute toxicity in rat liver. The amelioration of liver toxicity by the ethyl acetate extract was evident from its significant effect on the levels of serum glutamate pyruvate transaminase (SGPT), serum glutamate oxaloacetate transaminase (SGOT) and serum alkaline phosphatase (ALP). The results suggest that hepatoprotective effect of P. rimosus is possibly related to the free radical scavenging activity.

A new commercial strain of ergot adapted from a wild grass.

A strain of Claviceps purpurea (Fr.) Tul. isolated from Arthraxon lancifolius Hochst. was successfully adapted on rye. Experimental cultivation on rye showed 370 kg/ha yield of sclerotia containing 0.5% total alkaloid which consisted of 33% ergometrine, 17.6% ergotamine, 18.7% ergocornine and 22.7% ergokryptine. The results also reveal that alkaloid profile of the ergot strain depends largely on its host. Significantly high yield of the sclerotia and the valuable alkaloid profile of the new strain indicate its commercial potential.