Polysaccharide-Protein Complex Isolated from Fruiting Bodies and Cultured Mycelia of Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Agaricomycetes), Attenuates Doxorubicin-Induced Oxidative Stress and Myocardial Injury in Rats.

This study aimed to evaluate the effect of the polysaccharide-protein complex isolated from the fruiting bodies (GLFPPC) and cultured mycelia (GLMPPC) of a highly valued medicinal mushroom, Ganoderma lucidum, to alleviate doxorubicin (DOX)-induced cardiotoxicity. GLFPPC and GLMPPC were isolated from aqueous-alcoholic extracts of fruiting bodies and cultured mycelia of G. lucidum by repeated ethanol precipitation, dialysis, treatment with Sevag reagent, and freeze drying. The polysaccharide component was confirmed by assays with anthrone and phenol-sulphuric acid regents and protein moiety with Bradford reagent. The amino acid profile of protein moiety was determined by high-performance liquid chromatography analysis. DOX-induced cardiotoxicity was determined using Swiss albino mice. DOX administration caused a marked increase of creatine kinase and lactate dehydrogenase enzyme activities, indicating injury to the myocardium. The polysaccharide-protein complex downregulated cardiac injury marker enzymes, enhanced activities of endogenous antioxidants (namely, superoxide dismutase, catalase, glutathione peroxidase, and reduced glutathione levels), and significantly attenuated lipid peroxidation. The results indicated that GLFPPC and GLMPPC imparted protection against DOX-induced oxidative stress. Biochemical assays coupled with histopathological observations supported this conclusion. These experimental findings suggest that the polysaccharide-protein complex isolated from G. lucidum might be a useful therapeutic agent to ameliorate DOX-induced cardiomyopathy.

Morel mushroom, Morchella from Kashmir Himalaya: a potential source of therapeutically useful bioactives that possess free radical scavenging, anti-inflammatory, and arthritic edema-inhibiting activities.

Morel mushrooms, Morchella species are highly nutritious and excellently edible wild mushrooms abundantly growing in Kashmir Himalayas. The free radical scavenging, anti-inflammatory, and arthritis edema-inhibiting properties of bioactive extract of Morchella elata (EAE) were evaluated. EAE inhibited 53.2% formalin-induced paw edema at a dose of 500 mg/kg b.wt and 75.0% croton oil-induced skin inflammation at a dose of 50 mg topical application. EAE exhibited 51.8% COX inhibiting activity at a concentration of 100 µg/ml when assayed using LPS-stimulated RAW 264.7 cells exposed to the extract. NF-kB inhibiting activity of EAE was assayed using Lentix-293T P65 Ds Red stable cell line. High-throughput fluorescent imaging and flow cytometry showed profound ability of EAE to inhibit NF-kB activity. HPTLC analysis revealed that EAE is composed of several chemical components. The mushroom is a source of therapeutically useful functional food that can provide relief in arthritis.

Caterpillar Medicinal Mushroom, Cordyceps militaris (Ascomycetes), Mycelia Attenuates Doxorubicin-Induced Oxidative Stress and Upregulates Krebs Cycle Dehydrogenases Activity and ATP Level in Rat Brain.

Post-chemotherapy-induced cognitive dysfunction remains one of the challenges in cancer survivors. Cytokine-induced neurotoxicity manifests in subjects at any time after doxorubicin (DOX) chemotherapy. We examined the effect of bioactive Cordyceps militaris mycelia extract (CM) on the energy status, oxidative stress, and acetylcholinesterase activity in the brain of DOX treated rats. The CM (150 and 300 mg/kg b.w.) and DL-α lipoic acid (LA, 100 mg/kg b.w) were administered orally once daily for 5 days to male Wistar rats prior to the DOX administration (18 mg/kg as 3 doses of 6 mg/kg, i.p. b.w.) and continued for 6 more days. Cellular antioxidant status, Krebs cycle dehydrogenases, electron transport chain complexes (ETC) (I, III, and IV), adenosine triphosphate (ATP) level, advanced oxidation of protein products (AOPP), and acetylcholinesterase (AchE) activities were determined in the brain homogenate. The DOX alone treated group of animals showed significant decrease (p < 0.05) of brain antioxidant levels, Krebs cycle dehydrogenases activities, ETC complex activities, and decreased ATP level, while lipid peroxidation and AOPP levels were elevated. CM at 300 mg/kg b.w. or LA at 100 mg/kg b.w. elevated antioxidant status, Krebs cycle dehydrogenases, and complex activities and thus alleviated the toxicity. CM also inhibited the AchE activity in brain. The experimental results thus reveal that CM possessed excellent capacity to attenuate oxidative stress, upregulate respiratory chain complex activity and ATP levels, as well as inhibition of AchE activity.

Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Agaricomycetes), Prevents Doxorubicin-Induced Cardiotoxicity in Rats.

Doxorubicin (DOX) is an anticancer drug used extensively to treat a variety of human malignancies. DOX chemotherapy often leads to serious cardiotoxicity. We examined the ability of a Ganoderma lucidum extract (GLE) to prevent DOX-associated cardiotoxicity. DOX treatment of cardiac tissue drastically increased levels of creatine kinase (CK), lactate dehydrogenase (LDH), lipid peroxidation (thiobarbituric acid-reactive substances), advanced oxidation protein products (AOPPs), and protein carbonyls (PCOs), and significantly decreased reduced glutathione (GSH), superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase activities. Administration of GLE restored CK, LDH, AOPPs, and PCOs to almost normal levels and significantly enhanced the activity of SOD, GPx, catalase, and GSH; it also downregulated lipid peroxidation. Histopathological observations, hematology profiles, and electrocardiography parameters supported the protective effect of GLE against cardiotoxicity associated with DOX treatment.

Antiproliferative and Apoptotic Activities of the Medicinal Mushroom Phellinus rimosus (Agaricomycetes) on HCT116 Human Colorectal Carcinoma Cells.

Phellinus rimosus is a host-specific wood-rotting polypore that has been reported to be used by some tribes in Kerala, India, for curing mumps. We isolated a novel polysaccharide-protein complex from Ph. rimosus (PPC-Pr) that possessed significant antitumor activity. In this study, we examined the antiproliferative and apoptotic properties of PPC-Pr using human colon cancer cell line HCT116 as a model system. HCT116 cells were cultured in the presence of PPC-Pr at various concentrations (100-1000 µg/mL) for 24-96 hours, and the percentage of cell viability was evaluated using a 3-(4-5 dimethylthiozol-2-yl) 2-5-diphenyl-tetrazolium bromide (MTT) assay. The results showed that PPC-Pr inhibited cell viability in a time- and concentration-dependent manner. The antiproliferative effect of PPC-Pr was associated with apoptosis on HCT116 cells. We analyzed the morphological changes in the PPC-Pr-treated HCT116 cells using fluorescent DNA-microscopy with 4',6-diamidino-2-phenylindole dihydrochloride hydrate (DAPI) and acridine orange-ethidium bromide (AO/EB) staining. With DAPI staining, the cells treated with PPC-Pr showed nuclear shrinkage and chromatin condensation. Apoptotic morphology in cellular bodies as well as chromatin condensation were also confirmed by AO/EB double staining. Oligonucleosomal DNA fragmentation in PPC-Pr-treated cells was evaluated using a comet assay. The comet assay showed that control cells had few nuclei with fragmented chromatin, whereas apoptotic nuclei were more frequent in PPC-Pr-treated cells. These investigations indicate that the polysaccharide-protein complex from Ph. rimosus possesses antiproliferative activity and induces marked apoptosis in tumor cells in vitro.

Poly-MVA attenuates 7,12- dimethylbenz[a]anthracene initiated and croton oil promoted skin papilloma formation on mice skin.

OBJECTIVE: Chemopreventive agents which exhibit activities such as anti-inflammation, inhibition of carcinogen induced mutagenesis and scavenging of free radical might play a decisive role in the inhibition of chemical carcinogenesis either at the initiation or promotion stage. Many synthesized palladium (Pd) complexes tested experimentally for antitumor activity are found effective. Poly-MVA is a liquid blend preparation containing B complex vitamins, ruthenium with Pd complexed with alpha lipoic acid as the major ingredients. The antitumor effect of Poly-MVA was evaluated against 7,12-dimethylbenz[a] anthracene-initiated croton oil-promoted papilloma formation on mice skin. Skin tumor was initiated with a single application of 390 nmol of DMBA in 20 µl acetone. The effect of Poly-MVA against croton oil- induced inflammation and lipid peroxidation on the mice skin was also evaluated. Topical application of Poly-MVA (100 µl, twice weekly for 18 weeks) 30 minutes prior to each croton oil application, significantly decreased the tumor incidence (11%) and the average number of tumor per animals. Application of Poly-MVA (100 µl) before croton oil significantly (p &#60; 0.05) protected the mouse skin from inflammation (36%) and lipid peroxidation (14%) when compared to the croton oil alone treated group. Experimental results indicate that Poly-MVA attenuate the tumor promoting effects of croton oil and the effect may probably be due to its anti-inflammatory and antioxidant activity.

Antioxidant, Anti-Inflammatory, and Antitumor Activities of Cultured Mycelia and Fruiting Bodies of the Elm Oyster Mushroom, Hypsizygus ulmarius (Agaricomycetes).

Ethanoic extracts from the fruiting bodies and mycelia of the elm oyster mushroom, Hypsizygus ulmarius, were evaluated for their antioxidant, anti-inflammatory, and antitumor properties. Ethnolic extracts of fruiting body and mycelia showed 88%, 85%, 71%, and 85%, 65%, 70% 2,2-diphenyl-1-picrylhydrazyl, hydroxyl (DPPH) and 2,2'-azinobis (3-ethyl benzothiazolin-6-sulfonic acid) (ABTS) radical-scavenging activities, respectively, at a concentration of 1000 µg/mL. The anti-inflammatory activity was determined using carrageenan- and formalin- induced paw edema models. Diclofenac was used as the standard drug. In both models, the mycelia extract showed higher activity than the fruiting body extract. The antitumor effect of the extracts against Dalton's Lymphoma Ascites cell-line-induced tumors showed significant antitumor activity. Mycochemical analysis confirmed the presence of many pharmacologically active compounds such as phenol, alkaloids, proteins, tannins, and polysaccharides. Among these, polysaccharides and phenolic compounds were present at a higher concentration in both extracts. These compounds might be largely responsible for the mushroom's medicinal properties. The results of this study indicate that H. ulmarius possesses significant antioxidant, anti-inflammatory, and antitumor properties.

Hypoglycemic and Hypolipidemic Effects of the Cracked-Cap Medicinal Mushroom Phellinus rimosus (Higher Basidiomycetes) in Streptozotocin-Induced Diabetic Rats.

Phellinus rimosus is a parasitic host specific polypore mushroom with profound antioxidant, antihepatotoxic, anti-inflammatory, antitumor, and antimutagenic activities. This study investigated the hypoglycemic and hypolipidemic activities of the wood-inhabiting polypore mushroom Ph. Rimosus in streptozotocin (STZ)-induced diabetic rats. Diabetes was induced by single intraperitoneal injection of STZ (45 mg/kg) to Wistar rats. The effects of 30 days treatment with Ph. Rimosus (50 and 250 mg/ kg) and glibenclamide (0.65 mg/kg) on blood glucose level, serum insulin, serum lipid profile, liver glycogen, liver function enzymes, and non-enzymic and enzymic antioxidants activities in pancreas, liver, and kidney were evaluated in STZ-induced diabetic rats. Oral administration of Ph. Rimosus extract exhibited a significant reduction in blood glucose, triacylglycerol, total cholesterol, LDL-cholesterol, and liver function enzymes, and increased serum insulin, liver glycogen, and HDL-cholesterol levels in STZ-induced diabetic rats. Furthermore, Ph. Rimosus treatment increased antioxidant status in pancreas, liver, and kidney tissues with concomitant decreases in levels of thiobarbituric acid- reactive substances. Results of this study indicated that Ph. Rimosus possessed significant hypoglycemic and hypolipidemic activities and this effect may be related to its insulinogenic and antioxidant effect.

Medicinal Mushroom Cracked-Cap Polypore, Phellinus rimosus (Higher Basidiomycetes) Attenuates Acute Ethanol-Induced Lipid Peroxidation in Mice.

Alcohol abuse and alcoholism remain one of the major health issues worldwide, especially in developing countries. The protective effect of Phellinus rimosus against acute alcohol-induced lipid peroxidation in the liver, kidney, and brain as well as its effect against antioxidant enzyme activity such as superoxide (SOD) and catalase (CAT) in the liver was evaluated in mice. Ethyl acetate extract of Ph. Rimosus (50 mg/kg body wt, p.o.) 1 h before each administration of alcohol (3 mL/kg, p.o.; total 2 doses at 24-h intervals) protected against lipid peroxidation in all organs and attenuated the decline of SOD and CAT activity in the liver. The fold increase in lipid peroxidation, including conjugated diene and thiobarbituric acid reactive substance (TBARS) levels, was highest in the liver. There were 2.6- and 1.5- fold increases in TBARS levels in the liver of the alcohol alone- and alcohol+Ph. Rimosus-treated groups, compared with that of the normal group. Activity of SOD and CAT in the liver of alcohol- and alcohol+Ph. Rimosus- treated animals was 9.05±1.38, 18.76±1.71, and 11.26±1.02, 31.58±3.35 IU/mg protein, respectively. Extract at 1 mg/mL inhibited 50.6% activity of aniline hydroxylase (CYP2E1) in liver homogenate. From these results, we concluded that the extract significantly protected against the lipid peroxidation. Protection in the liver may be due to the inhibitory effect on CYP2E1 as well as the direct radical scavenging effect of Ph. Rimosus, which warrants further research.

The medicinal cracked-cap polypore mushroom Phellinus rimosus (higher Basidiomycetes) attenuates alloxan-induced hyperglycemia and oxidative stress in rats.

Diabetes is usually associated with increased production of reactive oxygen species (ROS), impaired antioxidant defense systems, or both, which result in oxidative damage and lead to ROS-mediated diabetic pathogenesis. This investigation was undertaken to evaluate the role of extract from the wood-inhabiting polypore medicinal mushroom Phellinus rimosus in an alloxan-induced diabetic model and the oral glucose tolerance test in rats. Oral administration of extract at doses of 50 and 250 mg/kg body weight/day for 10 days to rats with alloxan-induced diabetes was found to possess significant dose-dependent hypoglycemic activity. In the oral glucose tolerance test, hypoglycemic effect of P. rimosus (250 mg/kg) was significant (P < 0.01) and maximum at 90 minutes after the glucose challenge when compared with that of control group. The effect of extract on antioxidant status in the pancreas, liver, and kidney was estimated. The diabetic control rats exhibited elevated levels of lipid peroxidation and lower activities of copper/zinc superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and reduced glutathione (GSH) content in pancreatic, hepatic, and renal tissues compared with normal tissues. The activities of SOD, CAT, GPx, and GSH were found to be increased in diabetic rats treated with the extract. The increased level of lipid peroxidation in diabetic rats also was found to revert to near-normal status in groups treated with the extract. The findings thus suggest the therapeutic efficiency of Ph. Rimosus against declined antioxidant status as well as hyperglycemia associated with diabetes.

Antitumor activity of a polysaccharide-protein complex isolated from a wood-rotting polypore macro fungus Phellinus rimosus (Berk) Pilat.

A protein-bound, water-soluble polysaccharide-protein complex was isolated from a medicinal mushroom, Phellinus rimosus (Berk) Pilat (PPC-Pr). The isolation was achieved by hot water extraction, filtration, solvent precipitation, dialysis, and freeze-drying. The proximate analysis showed that PPC-Pr comprised 54.8% polysaccharide and 28.6% protein. The molecular weight of the compound was determined by gel filtration using a Sephadex G 100. The molecular weight of PPC-Pr was approximately 1,200,000 D. The thin-layer chromatography analysis of PPC-Pr after acid hydrolysis with trifluroacetic acid showed that it was composed of glucose as the only monosaccharide unit. The amino acid profile analysis of PPC-Pr revealed that it contained large amounts of aspartic acid, glutamic acid, alanine, glycine, and serine. Thus, the results indicated that PPC-Pr is a glucan-protein complex. The PPC-Pr did not show in vitro cytotoxic activity against Dalton's lymphoma ascites and Ehrlich's ascites carcinoma cell lines. The PPC-Pr was found to be effective in increasing the life span of ascites tumors induced by Ehrlich's ascites carcinoma cell line in mice. PPC-Pr also was found to have significant preventive and curative effects on solid tumors induced by the Dalton's lymphoma ascites cell line. The experimental results thus indicated that protein-bound polysaccharide (PPC-Pr) isolated from P. rimosus possessed profound antitumor activity. The findings suggest the potential therapeutic use of this compound as an antitumor agent.

Palladium-α-lipoic acid complex attenuates alloxan-induced hyperglycemia and enhances the declined blood antioxidant status in diabetic rats.

BACKGROUND: Palladium α-lipoic acid (Pd-LA) complex has unique electronic and redox properties that appear to be the key to its physiological effectiveness. A proprietary liquid blend containing Pd-LA as the major component was demonstrated to be effective in improving the activities of mitochondrial enzymes in aged rats. METHODS: The Pd-LA complex was evaluated for its hypoglycemic effect against the alloxan-induced diabetic model, as well as in the oral glucose tolerance test in rats. The in vitro free radical scavenging activity of Pd-LA was also determined. RESULTS: Administration of Pd-LA (0.5 mL/kg; equivalent to 3.8 mg complexed α-lipoic acid/kg, p.o.) daily for 5 days to alloxan-induced diabetic animals significantly reduced the blood glucose level (P < 0.05). The blood antioxidant status in the diabetic animals was significantly improved by the treatment of Pd-LA (P < 0.05). Similarly, Pd-LA showed significant in vitro antioxidant activity in a concentration-dependent manner. CONCLUSIONS: Results of the study conclude that the Pd-LA complex is effective in lowering the blood glucose level and enhancing the declined antioxidant status in diabetic animals. Significant finding(s) of the study include: (i) Pd-LA significantly increased the tolerance of glucose and was also effective in ameliorating hyperglycemia induced by alloxan; (ii) Pd-LA significantly enhanced the activities of blood superoxide dismutase, catalase, glutathione peroxidase and level of glutathione in diabetic animals; and (iii) Pd-LA showed significant in vitro antioxidant activity. This study adds: The therapeutic efficiency of Pd-LA is demonstrated against declined antioxidant status as well as hyperglycemia associated with diabetes.

Free-radical scavenging and mitochondrial antioxidant activities of Reishi-Ganoderma lucidum (Curt: Fr) P. Karst and Arogyapacha-Trichopus zeylanicus Gaertn extracts.

Endogenous damage to mitochondrial DNA by free radicals is believed to be a major contributory factor to aging. The current study examined the effects of the extracts of two important anti-fatigue and rejuvenating medicinal herbs Ganoderma lucidum and Trichopus zeylanicus for their free-radical scavenging property and for their effects on liver mitochondrial antioxidant activity in aged mice. Both extracts were administrated orally to aged BALB/c mice at doses of 50 and 250 mg/kg body weight for 15 days. Super oxide dismutase (SOD) and catalase (CAT) activity and levels of reduced glutathione (GSH) and lipid peroxidation as equivalents of malondialdehyde (MDA) formed were determined. Groups of young mice and aged mice (more than 15 months old) were taken as controls. Both G. lucidum and T. zeylanicus extracts increased antioxidant status in liver mitochondria of aged mice compared with the aged control. Higher levels of GSH, increased activity of SOD and CAT, and decreased level of MDA in both treated groups compared with the controls were evident. Both extracts possessed significant 2,2-diphenyl-1-picrylhydrazil (DPPH), 2, 2'-azinobis (3-ethylbenzothiazolin-6-sulphonic acid) (ABTS) radical scavenging activities and ferric reducing antioxidant power (FRAP). The DPPH, ABTS, and FRAP activities were higher in G. lucidum extract than in T. zeylanicus. G. lucidum extract also showed superoxide and hydroxyl radical scavenging activities. T. zeylanicus had significantly higher lipid peroxidation inhibiting activity than G. lucidum. Thus, we conclude that the antioxidative effect of the G. lucidum extract was higher than that of T. zeylanicus. Our findings suggest a potential therapeutic efficacy of G. lucidum extract to protect against aging and to a certain extent against age-related degenerative diseases.