Analyses of Elaeocarpus sphaericus Extract for Antioxidant, Antiproliferative and Gene Repression Activities against HIF-1α and VEGF.

The study presents antioxidant, phytochemical, anti-proliferative, and gene repression activities against Hypoxia-inducible factor (HIF-1) alpha and Vascular endothelial growth factor (VEGF) of Elaeocarpus sphaericus extract. Elaeocarpus sphaericus dried and crushed plant leaves were extracted using water and methanol by ASE (Accelerated Solvent Extraction) method. Total phenolic content (TPC) and total flavonoid content (TFC) were used to measure the extracts' phytochemical activity (TFC). Antioxidant potential of the extracts was measured through DPPH, ABTS, FRAP, and TRP. Methanolic extract of the leaves of E. sphaericus has shown a higher amount of TPC (94.666±4.040 mg/gm GAE) and TFC value (172.33±3.21 mg/gm RE). The antioxidant properties of extracts in the yeast model (Drug Rescue assay) showed promising results. Ascorbic acid, gallic acid, hesperidin, and quercetin were found in the aqueous and methanolic extracts of E. sphaericus at varying amounts, according to a densiometric chromatogram generated by HPTLC analysis. Methanolic extract of E. sphaericus (10 mg/ml) has shown good antimicrobial potential against all bacterial strains used in the study except E. coli. The anticancer activity of the extract in HeLa cell lines ranged from 77.94±1.03 % to 66.85±1.95 %, while it ranged from 52.83±2.57 % to 5.44 % in Vero cell lines at varying concentration (1000 µg/ml-31.2 µg/ml). A promising effect of extract was observed on the expression activity of HIF-1 and VEGF gene through RT-PCR assay.

In vitro antioxidant profiling of seabuckthorn varieties and their adaptogenic response to high altitude-induced stress.

In the past few years, seabuckthorn plants have gained special attention due to their ability to grow in the harshest of the environment. This adaptability may be contributed by various antioxidants present in the plants besides other morphological adaptation. As in vivo studies cannot be justified without in vitro studies, the present investigation carried out evaluation of both in vitro and in vivo antioxidant potentials of aqueous and alcoholic extracts of the leaves of Hippophae salicifolia (HS) and Hippophae rhamnoides mongolica (HRM) in comparison with Hippophae rhamnoides turkestanica (HRT). The results had clearly depicted that in vitro antioxidant potential of the extracts was responsible for the in vivo adaptogenic performance in animals during cold and hypoxia exposure under restraint stress. Total phenolic content (TPC), total flavonoid content (TFC), total protein content, and antioxidant potential were determined. For adaptogenic studies, rats with oral drug supplementation were exposed to Cold-hypoxia-restraint (C-H-R) stresses-induced hypothermia, as a measure of endurance. Aqueous extracts of HS showed maximum (99 %) resistance compared to HRT (81 %) and HRM (29 %). The levels of biochemical parameters such as malondialdehyde (MDA), reactive oxygen species (ROS), lactate dehydrogenase (LDH), superoxide dismutase (SOD), glutathione (GSH/GSSG), and catalase (CAT) in blood samples also revealed that the aqueous leaf extract of HS has better antioxidant and adaptogenic potential compared to HRM.

Phytochemical and antimicrobial activities of Himalayan Cordyceps sinensis (Berk.) Sacc.

This study evaluated the phytochemical and antimicrobial activities and also quantified bioactive nucleoside using high performance thin layer chromatography (HPTLC) of five extracts of Indian Himalayan Cordyceps sinensis prepared with different solvents employing accelerated solvent extraction (ASE) technique. The phytochemical potential of these extracts was quantified in terms of total phenolic and total flavonoid content while antioxidant activities were determined by 1,1-diphenyl-2-pycryl-hydrazyl (DPPH) and 2,2 -azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and ferric-reducing antioxidant power (FRAP) assays. Total reducing power (TRP) was determined by converting iron (III) into iron (II) reduction assay. CS(50%Alc) (15.1 ± 0.67mg/g of dry extract) and CS(100%Alc) (19.3 ± 0.33 mg/g of dry extract) showed highest phenolic and flavonoid content, respectively while CS(Aq) extract showed maximum antioxidant activity and the highest concentration of the three nucleosides (adenine 12.8 ± 0.49 mg/g, adenosine 0.36 ± 0.28 mg/g and uracil 0.14 ± 0.36 mg/g of dry extract) determined by HPTLC. The evaluation of extracts for antimicrobial activity against gram-negative and gram-positive bacterial strains showed CS(25%Alc), CS(75%Alc) and CS(100%Alc) extract to be more effective against E. coli, P. aerugenosa and B. subtilis giving 9, 7 and 6.5 mm of zone of inhibition (ZOI) in 93.75, 93.75 and 45 µg concentration, respectively, whereas CS(Aq) extract showed minimal inhibition against these.

Subcritical water extraction of antioxidant compounds from Seabuckthorn (Hippophae rhamnoides) leaves for the comparative evaluation of antioxidant activity.

A novel environmentally friendly technique, subcritical water extraction (SWE) was employed for the extraction of antioxidant compounds from Seabuckthorn leaves (SBT). Antioxidant activity of the extracts was evaluated using commonly accepted chemical assays. Also, present study reports the cytoprotective and antioxidant properties of SBT against tertiary-butyl hydroperoxide (tert-BOOH) induced oxidative stress in murine macrophages (Raw 264.7). Exposure of cells to tert-BOOH resulted, increase in cytotoxicity, reactive oxygen species (ROS) production and decrease in mitochondrial membrane potential, which is responsible for fall in intracellular antioxidant levels. Pretreatment of cells with SBT extracts inhibited cytotoxicity, ROS production and maintained antioxidants levels similar to that of control cells. The chemical composition of the SWE extracts studied showed total phenol content (76.07-93.72mg/g GAE) and total flavonoid content (47.06-66.03mg/g rutin). Further, some of its phenolic constituents; (1) Quercetin-3-galactoside, (2) Kaempferol and (3) Isorhamnetin were quantified by RP-HPLC.

Supercritical Carbon Dioxide Extracts of Cordyceps sinensis: Chromatography-based Metabolite Profiling and Protective Efficacy Against Hypobaric Hypoxia.

The toxicity and disposal concerns of organic solvents used in conventional extraction purposes has entailed the need for greener alternatives. Among such techniques, supercritical fluid extraction (SFE) has gained popularity by yielding extracts of high purity in a much faster manner. Carbon dioxide (CO2) is generally preferred as a supercritical solvent because of its lower temperature requirements, better diffusivity and easy removal. The present study describes the characterization of supercritical CO2 extracts of Indian variety of Cordyceps sinensis (CS)- a high-altitude medicinal mushroom widely revered in traditional medicine for its extensive anti-hypercholesterolemic, anti-inflammatory, anti-proliferative and energy-enhancing properties. Experimental parameters viz. 300 and 350 bar of extraction pressure, 60°C of temperature, 0.4°L/h CO2 of flow rate and use of 1% (v/v) of ethanol as entrainer were optimized to prepare three different extracts namely, CSF1, CSF2 and CSF3. High-performance thin-layer chromatography (HPTLC) was used for assessing the quality of all the extracts in terms of cordycepin, the pivot biomarker compound in CS. Characterization by HPTLC and GC-MS confirmed the presence of flavonoids and nucleobases and, volatile organic compounds (VOCs), respectively. The chromatographic data acquired from metabolite profiling were subjected to chemometric analysis in an open source R studio which illustrated interrelatedness between CSF1 and CSF2 in terms of two major principal components. i.e. Dim 1 and Dim 2 whose values were 40.33 and 30.52% in variables factor map plotted using the HPTLC-generated retardation factor values. The factor maps based on retention times of the VOCs exhibited a variance of Dim 1 = 43.95% and Dim 2 = 24.85%. Furthermore, the extracts demonstrated appreciable antibacterial activity against Escherichia coli and Salmonella typhi by generation of reactive oxygen species (ROS), protein leakage and efflux pump inhibition within bacterial pathogens. CSFs were elucidated to be significantly cytoprotective (p < 0.05) in a simulated hypobaric hypoxia milieu (0.5% oxygen). CSF2 showed the best results by effectively improving the viability of human embryonic kidney (HEK 293) cells to 82.36 ± 1.76% at an optimum dose of 100 µg/ml. Levels of hypoxia inducible factor-1 alpha (HIF-1α) were modulated four-fold upon supplementation with CSF2. The results collectively evinced that the CSF extracts are substantially bioactive and could be effectively utilized as mycotherapeutics for multiple bioeffects.

Novel formulation development from Ophiocordyceps sinensis (Berk.) for management of high-altitude maladies.

Ophiocordyceps sinensis (Berk.) is a fungus closely related to medicinal mushroom, which belongs to the family Ophiocordycipitaceae. It is a well-known and rich herbal source of bioactive active constituents. The medicinal mushroom has garnered worldwide attention owing to its multifarious bioactivities. This mushroom grows on the larva of ghost moths (Hepialidae) and produces fruiting bodies, which serve as a vital natural source of medicine and supplementary diets. On account of the diverse pharmacological and bioactive constituents present in O. sinensis, it has been established as a potential antioxidant, anticancer, antibacterial, anti-proliferative, anti-inflammatory agent that has been successfully used for treating several health issues, including hypoxia-related problems encountered by mountaineers, pilgrims, tourists and soldiers occurring at high-altitude regions such as acute mountain sickness (AMS), high-altitude pulmonary edema (HAPE), high-altitude cerebral edema (HACE), frostbite, chilblains, hypothermia, etc. The most important pharmacologically active compounds present in the O. sinensis include nucleobases and its derivatives (adenosine, cordycepin, 3-deoxyadenosine, AMP, GMP, UMP, guanosine, uridine), polysaccharides (mannose, glucose, galactose, rhamnose, arabinose, xylose, galactose), proteins, peptides and steroids. This article focuses on the various research endeavors undertaken to scientifically establish the medicinal properties of O. sinensis, highlighting the various principally active compounds, their pharmacological action, drug designing and development and future perspective for various health benefits.

Effects of Extraction Temperature on Efficacy of Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Agaricomycetes), Aqueous Extract against Oxidative Stress.

This study was designed to understand the effect of extraction temperature, i.e., room temperature (GLRT), 50°C (GL50), 100°C (hot water; GL100), and 200°C (GL200) on antioxidant and biological activity of G. lucidum. The % yield obtained was 5.3%, 7.6%, 10.7%, and 13.2% at various extraction temperatures; room temperature, 50°C, 100°C and 200°C, respectively. Similarly, phenolic content (51.6, 57.9, 82.9, and 93.1 mg/g extract) and flavonoid content (18.8, 23.2, 34.3, and 36.3 mg/g extract) were observed to be increased with rise in extraction temperature. However, extraction temperature resulted in loss of antioxidant activities above 100°C as evident by chemical assays such as DPPH, FRAP, ABTS, and TRP conducted on extracts. In contrast, three bioactive compounds, i.e., adenine (3.26, 3.48, 2.16, and 1.45 mg/g extract), uracil (3.99, 3.21, 2.51, and 1.47 mg/g extract), and adenosine (5.92, 5.62, 2.22 and 0.7 mg/g extract), quantified by high performance thin layer chromatography showed decrease in their content with increasing extraction temperature. Extract prepared at room temperature and 50°C prevented loss of cell viability and generation of reactive oxygen species resulted after hydrogen peroxide exposure; however, cytoprotective efficacy was not significant at 100°C and 200°C The order of cytoprotective effects observed by these extract were in the following order: room temperature ≥ 50°C > 100°C > 200°C. Overall, the optimal temperature conditions for the efficient extraction of G. lucidum with water retaining bioactive compounds and biological activity was found to be below 100°C.

Bioactive Fractions from the Chinese Caterpillar Mushroom, Ophiocordyceps sinensis (Ascomycetes), Elucidate Adaptogenic Role against Hypoxia Stress.

Ophiocordyceps sinensis, a high-altitude medicinal mushroom, is widely revered in traditional medicine for its antiproliferative, antihypercholesterolemic, energy enhancement, etc. properties. These properties are attributed to the presence of steroids, terpenoids, polyphenolics, glycosides, and glycoproteins in it. The current study presents characterization of three phenolic rich fractions (PRFs) separated from aqueous extract of O. sinensis using diethyl ether and ethyl acetate. These fractions possessed considerable antioxidant potential in terms of total phenolic content, total flavonoid content, ferric ion reducing power, and free radical scavenging efficiency. Chemical characterization of the PRFs was carried out using HPTLC and GC-MS. The diethyl ether and ethyl acetate fractions elucidated appreciable antibacterial activity against Salmonella typhi. The aqueous extract and residual phenolic fraction displayed protective effect on HEK 293 cell lines against hypoxia stress and also bestowed appreciative adaptogenic role against cold, hypoxia, and restraint stress in vivo, by decreasing levels of malondialdehyde and superoxide dismutase and incrementing reduced glutathione concentrations.

Antioxidant-Rich Peptide Fractions Derived from High-Altitude Chinese Caterpillar Medicinal Mushroom Ophiocordyceps sinensis (Ascomycetes) Inhibit Bacterial Pathogens.

Ophiocordyceps sinensis (=Cordyceps sinensis), a medicinal mushroom native to the Orient, has been extensively used for the past few centuries in traditional Chinese medicine because of its immunomodulatory, antiinflammatory, and nutraceutical properties. In the present study, three antioxidant and antibacterial Ophiocordyceps peptide fractions (COPs) were separated from the Indian variety of O. sinensis on a Sephadex G-25 resin. Amide bonds in the COPs were confirmed by Fourier transform infrared spectroscopy and nuclear magnetic resonance. Cationic and hydrophobic amino acids, which are reported to be the major constituent amino acids of antimicrobial peptides, were identified in the COPs by matrix-assisted laser desorption/ionization-mass spectrometry and high-performance liquid chromatography. Putative secondary structures were predicted by circular dichroism to be ß-sheets and random coils. The COPs demonstrated substantial antioxidant potential by scavenging 2,2-diphenyl-l-picryl-hydrazyl-hydrate (median inhibitory concentration [IC50] values, 4.79-18.7 mg/mL) and 2,2'-azino-bis-3-ethylbenzothiazoline-6-sul-phonic acid (IC50 values, 4.51-14.05 mg/mL) free radicals and also by chelating heavy metal ions. Additionally, the peptide fractions were capable of significantly inhibiting bacterial pathogens viz. Escherichia coli and Salmonella typhi. The potential antibacterial mechanisms of action were established to be generation of reactive oxygen species and intracellular protein leakage within the bacterial cells.

Role of Hydroalcoholic Extract of Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Agaricomycetes), in Facilitating Cellular Acclimatization in a Low-Oxygen Microenvironment.

Ganoderma lucidum is known to exert many health benefits including effects to improve oxygen utilization. Therefore, this study was designed to evaluate the role of a hydroalcoholic G. lucidum extract in providing tolerance to HT22 cells grown under hypoxic conditions. HT22 cells were exposed to 0.5% O2 in the presence or absence of the extract for 24 hours. At the end of the exposure period, we performed cell viability assays, cell cycle analysis, and biochemical and protein expression studies. The extract-treated cells revealed less cell death, minimized caspase 3 and reactive oxygen species levels, and relieved G0/G1 cell cycle arrest compared with hypoxic cells cultured without the extract. Further, extract-treated cells showed improved expression of Nrf2, heme oxygenase 1, and metallothionein and stabilized levels of hypoxia-inducible factor 1α. Moreover, lower levels of nuclear factor-κB and tumor necrosis factor a were evident in extract-treated cells. Overall, the G. lucidum extract reduced hypoxia-induced cell death and augmented transcription factors (HIF-1α and Nrf2), conferring tolerance to hypoxia.

Phenolic Rich Fractions from Mycelium and Fruiting Body of Ganoderma lucidum Inhibit Bacterial Pathogens Mediated by Generation of Reactive Oxygen Species and Protein Leakage and Modulate Hypoxic Stress in HEK 293 Cell Line.

Ganoderma lucidum (G. lucidum) fungus (Family Ganodermataceae) is widely used as a traditional medicine in China, Japan, and many Asian countries on account of its numerous medicinal properties such as antioxidant, anticancer, antimicrobial, energy enhancing, and immunostimulatory. This broad spectrum of therapeutic effects exhibited by G. lucidum is ascribed to its abundance in several classes of chemical constituents, namely, carbohydrates, flavonoids, minerals, phenolics, proteins, and steroids which possess substantial bioactivities. The aim of the current study was to prepare phenolic rich fractions (PRFs) from aqueous extract of the Indian variety of G. lucidum mycelium and fruiting body. These fractions were assessed for their antioxidant capacity by TPC (total phenolic content), TFC (total flavonoid content), FRAP (ferric reducing antioxidant power), and ABTS [2,2-azino-bis(3-ethylbenzothiazoline)-6-sulfonic acid] assays. Quantification of flavonoids and nucleobases present in the fractions was carried out by high-performance thin layer chromatography (HPTLC). The antibacterial activity of the fractions was evaluated against Escherichia coli, Salmonella typhi, and Staphylococcus aureus. The antibacterial mechanism of action of the PRFs was established to be generation of reactive oxygen species and leakage of proteins within bacterial cells. Additionally, the protective effect of the PRFs in counteracting hypoxia was observed in HEK 293 cell lines.

Valeriana wallichii extract inhibits tert-BOOH induced oxidative damage and cytotoxicity.

Oxidative stress is well known to be involved in pathophysiology of several disorders. Valeriana wallichii (VW) root extracts is known for its expedient activities but its antioxidant and cytoprotective efficacy need to be explored further. In present study tertiary-butyl hydroperoxide (tert-BOOH) was used to induce oxidative stress in C6 glioma cells. Antioxidant activity of the VW root extracts were evaluated by chemical assays i.e. DPPH, ABTS and FRAP assay. Further effect of VW on tert-BOOH induced oxidative stress and mitochondrial damage was studied. Result of present study revealed that exposure of cells to tert-BOOH resulted in increase in cytotoxicity, reactive oxygen species (ROS) production and decrease in mitochondrial membrane potential, super oxide dismutase, catalase, glutathione peroxidase and glutathione reductase. On the other hand, pretreatment of cells with VW extracts ameliorated these damaging effects. Additionally, HPLC analysis revealed hesperidin as an active ingredient and concentration of heavy metal was found within the maximum permissible limits prescribed by WHO. In conclusion present study revealed the antioxidant and cytoprotective property of VW against oxidative stress.

Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Agaricomycetes), as a Cardioprotectant in an Oxygen-Deficient Environment.

Imbalanced oxygen availability is detrimental to normal cell function. Oxygen-sensitive cells such as cardiomyoblasts experience severe irreversible pathophysiological damage under conditions of reduced oxygen availability, such as hypoxia. A number of natural therapeutic agents have been explored for their potential cytoprotective effects, of which medicinal mushrooms are an important source. Ganoderma lucidum, commonly known as lingzhi, is one such mushroom that has been elaborately studied for its potential pharmacological properties. In this study, aqueous and alcoholic extracts of a natural Himalayan variety of G. lucidum were evaluated for their efficiency as remedial agents in treating hypoxic injury to H9c2 cardiomyoblasts. The alcoholic extract of G. lucidum effectively restored cellular viability at a concentration of 600 µg/mL and aided in maintaining cellular redox balance under hypoxia. Substantial reduction in caspase-3 and -7 activation was observed with fluorescent-activated cell sorting. Alcoholic extract of G. lucidum minimized oxidative stress as indicated by measuring reactive oxygen species, lipid peroxidation, and reduced glutathione-to-oxidized glutathione ratio, and also by determining changes in hypoxia-inducible factor 1α and associated genes. To ascertain these positive outcomes of administration of G. lucidum extracts, certain phytoconstituents (nucleobases and flavonoids) were identified using high-performance thin-layer chromatography; antioxidant potential was also evaluated. Results indicated that both extracts contained notable quantities of nucleobases and flavonoids. The extracts also effected high free radical scavenging activities.

Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Agaricomycetes), Inhibits Candida Biofilms: A Metabolomic Approach.

This article presents a comparative gas chromatography (GC)-mass spectrometry (MS)-based metabolomic analysis of mycelia and fruiting bodies of the medicinal mushroom Ganoderma lucidum. Three aqueous extracts-mycelia, fruiting bodies, and a mixture of them-and their sequential fractions (methanolic and ethyl acetate), prepared using an accelerated solvent extractor, were characterized by GC-MS to determine volatile organic compounds and by high-performance thin-layer chromatography to quantify ascorbic acid, a potent antioxidant. In addition, these extracts and fractions were assessed against Candida albicans and C. glabrata biofilms via the XTT reduction assay, and their antioxidant potential was evaluated. Application of chemometrics (hierarchical cluster analysis and principal component analysis) to GC data revealed variability in volatile organic compound profiles among G. lucidum extracts and fractions. The mycelial aqueous extract demonstrated higher anti-Candida activity and ascorbic acid content among all the extracts and fractions. Thus, this study illustrates the preventive effect of G. lucidum against C. albicans and C. glabrata biofilms along with its nutritional value.

Screening of Indian Lingzhi or Reishi Medicinal Mushroom, Ganoderma lucidum (Agaricomycetes): A UPC2-SQD-MS Approach.

Oriental medicinal mushroom Ganoderma lucidum has been widely used for the promotion of health and longevity owing to its various bioactive constituents. Therefore, comprehending metabolomics of different G. lucidum parts could be of paramount importance for investigating their pharmacological properties. Ultra-performance convergence chromatography (UPC2) along with mass spectrometry (MS) is an emerging technique that has not yet been applied for metabolite profiling of G. lucidum. This study has been undertaken to establish metabolomics of the aqueous extracts of mycelium (GLM), fruiting body (GLF), and their mixture (GLMF) using ultra-performance convergence chromatography single quadrupole mass spectrometry (UPC2-SQD-MS). Aqueous extracts of G. lucidum prepared using an accelerated solvent extraction technique have been characterized for their mycochemical activities in terms of total flavonoid content, 1,1-diphenyl-2-picryl-hydrazyl scavenging activity, and ferric ion reducing antioxidant power. The UPC2-SQD-MS technique has been used for the first time for metabolite profiling of G. lucidum on a Princeton Diol column (4.6 × 250 mm; 5 µm) using supercritical CO2 (solvent) and 20 mM ammonium acetate in methanol (co-solvent). In the present study, UPC2-SQD-MS was found to be a rapid, efficient, and high-throughput analytical technique, whose coupling to principal component analysis (PCA) and phytochemical evaluation could be used as a powerful tool for elucidating metabolite diversity between mycelium and fruiting body of G. lucidum. PCA showed a clear distinction in the metabolite compositions of the samples. Mycochemical studies revealed that overall GLF possessed better antioxidant properties among the aqueous extracts of G. lucidum.

Protective Efficacy of the Caterpillar Mushroom, Ophiocordyceps sinensis (Ascomycetes), from India in Neuronal Hippocampal Cells against Hypoxia.

This study demonstrated the protective efficiency of extracts of the Indian variety of Ophiocordyceps sinensis (=Cordyceps sinensis) (CSEs) in HT22 (murine hippocampal) cells under hypoxic conditions. Various parameters such as cell viability, reactive oxygen species, levels of endogenous antioxidants, inflammatory cytokines, transcription factors, and oxidation of macromolecules were analyzed. In addition, the radical scavenging abilities of hydroxyl radicals, nitric oxide, and superoxide radicals were also studied. Antioxidant compounds, ascorbic acid, hesperidin, and rutin were quantified by high-performance thin-layer chromatography. The information acquired from high-performance thin-layer chromatography profiling was subjected to principal component analysis for data clustering. Findings of this research revealed that ascorbic acid and rutin were highest in aqueous CSE, whereas the maximum amount of hesperidin was found in 25% alcoholic CSE. In vitro studies showed that all the CSEs protected HT22 cells well by upregulating the level of endogenous antioxidants and preventing the oxidation of lipids and proteins. These extracts also reduced the amount of hypoxia-induced inflammatory cytokines and transcription factors on par with the normoxic control with more or less equal protection in the cells under hypoxia, and indicated significant radical scavenging potential.

Wound healing activity of an aqueous extract of the Lingzhi or Reishi medicinal mushroom Ganoderma lucidum (higher Basidiomycetes).

The Lingzhi or Reishi medicinal mushroom Ganoderma lucidum (higher Basidiomycetes) is popular because of its health-promoting properties. The effects of G. lucidum extract on cancer, hypertension, hypercholesterolemia, and hepatitis have been reported by many researchers. This investigation was undertaken to evaluate the healing efficacy of an aqueous lyophilized extract of G. lucidum from the Indian Himalayan region on dermal excision wound in experimental rats. The extract used in the study was found to be rich in total polyphenol and flavonoid contents. The healing efficacy was comparatively assessed with a reference povidone-iodine ointment. The G. lucidum extract showed significant enhanced healing activity, evidenced by an increase in wound contraction, collagen accumulation (hydroxyproline), hexosamine, and total protein contents. Histopathological findings further supported the biochemical indices. The results suggest that aqueous lyophilized extract of G. lucidum possesses significant wound-healing activity.

Scientific validation of the Chinese caterpillar medicinal mushroom, Ophiocordyceps sinensis (Ascomycetes) from India: immunomodulatory and antioxidant activity.

In the present study, we aimed to elucidate the antioxidant property and anti-inflammatory activity of the aqueous extract of the Indian species of Ophiocordyceps sinensis (AECS), which demonstrates medicinal activity against numerous diseases. The chemical composition of AECS was quantified using a colorimeteric technique to determine the total phenolic and flavonoid contents. Antioxidant activity was determined by assays for 2,2'-azinobis(3-ethylbenzothiazoline-6-sulphonic acid)diammonium salt (ABTS); 2,2-diphenyl-1-picryl-hydrazyl (DPPH); and ferric reducing antioxidant power (FRAP). Adenosine nucleoside and nitrogenous bases (adenine and uracil) were also quantified by high-performance thin layer chromatography (HPTLC). Furthermore, the aqueous extract was also analyzed for anti-inflammatory activity in vitro using THP1 cells. THP1 cells were treated with and without lipopolysaccharide (LPS) and AECS (at 25 µg/mL, 50 µg/mL and 100 µg/mL, respectively) for 24 h. After 24 h, supernatants were harvested and kept at -80°C until the cytokine assays were performed. Furthermore, nitric oxide (NO) content was also estimated in treated and untreated murine peritoneal macrophages using Griess reagent. AECS significantly suppressed LPS-induced release of TNF-α and IL-1ß in THP1 cells and significantly suppressed NO release in macrophage cells without exerting any toxic effect. These results indicate the anti-inflammatory activity of AECS. Additionally, this extract also has an antioxidant property, as high contents of phenols and flavonoids are present in the extract with considerable reducing power. The results of this study clearly demonstrate the potent antioxidant property and anti-inflammatory activity of AECS. Therefore, consumption of AECS may be clinically useful to protect against inflammatory diseases.

Efficacy of aqueous extract of Hippophae rhamnoides and its bio-active flavonoids against hypoxia-induced cell death.

OBJECTIVES: To investigate the protective efficacy of aqueous extract of Hippophae rhamnoides against chronic hypoxic injury using primary rat hepatocytes. MATERIALS AND METHODS: The extract was prepared using maceration method and characterized by its phenolic and flavonoid content and chemical antioxidant capacity using ferric reducing antioxidant power assay. Hepatocytes were maintained in hypoxia chamber (3% and 1% oxygen) for 72 h. The cells kept under normoxic condition served as control. The cells were treated with the extract and flavonoids; isorhamentin, kaempferol or qurecetin-3-galactoside. After the end of exposure period; cell survival, reactive oxygen species (ROS), leakage of lactate dehydrogenase (LDH), alanine aminotransferase (ALT), aspartate aminotransferase (AST), reduced glutathione (GSH), glutathione peroxidase (GPx), and superoxide dismutase (SOD) levels were measured. RESULTS: The extract showed presence of high phenolic and flavonoid content with significant antioxidant activity in chemical assay. The cell exposed to hypoxia showed concentration dependent cell death and harbored higher reactive oxygen species. In addition, these cells showed significant leakage of intracellular LDH, ALT, and AST accompanied by the diminished levels/activities of GSH, GPx, and SOD. The treatment of cells with aqueous extract of H. rhamnoides reduced hypoxia-induced cell death and prevented increase in ROS levels and leakage of intracellular LDH, ALT, and AST from cells. Moreover, these cells maintained better levels/activities of GSH, GPx, and SOD in comparison to the respective controls. The major flavonoids present in aqueous extract of H. rhamnoides; quercetin-3-galactoside, kaempferol, and isorhamentin also prevented hypoxia induced cell injury individually or in combination, however, the protection offered by these compounds taken together could not match to that of the extract. CONCLUSIONS: Overall the findings reveal significance of aqueous extract of H. rhamnoides in controlling ROS-meditated hypoxic injury in cells and can be useful in many hepatic complications.

Cordyceps sinensis increases hypoxia tolerance by inducing heme oxygenase-1 and metallothionein via Nrf2 activation in human lung epithelial cells.

Cordyceps sinensis, an edible mushroom growing in Himalayan regions, is widely recognized in traditional system of medicine. In the present study, we report the efficacy of Cordyceps sinensis in facilitating tolerance to hypoxia using A549 cell line as a model system. Treatment with aqueous extract of Cordyceps sinensis appreciably attenuated hypoxia induced ROS generation, oxidation of lipids and proteins and maintained antioxidant status similar to that of controls via induction of antioxidant gene HO1 (heme oxygenase-1), MT (metallothionein) and Nrf2 (nuclear factor erythroid-derived 2-like 2). In contrast, lower level of NF κ B (nuclear factor kappaB) and tumor necrosis factor- α observed which might be due to higher levels of HO1, MT and transforming growth factor- ß . Further, increase in HIF1 (hypoxia inducible factor-1) and its regulated genes; erythropoietin, vascular endothelial growth factor, and glucose transporter-1 was observed. Interestingly, Cordyceps sinensis treatment under normoxia did not regulate the expression HIF1, NF κ B and their regulated genes evidencing that Cordyceps sinensis per se did not have an effect on these transcription factors. Overall, Cordyceps sinensis treatment inhibited hypoxia induced oxidative stress by maintaining higher cellular Nrf2, HIF1 and lowering NF κ B levels. These findings provide a basis for possible use of Cordyceps sinensis in tolerating hypoxia.