Inonotus Obliquus Research Articles

Inhibitory activity of tea compositions and their constituent ingredients on SARS‐COV‐2 replication in vitro

Aim. In vitro analysis of the inhibitory activity of aqueous extracts of ready‐made tea compositions and their constituent ingredients on SARSCOV‐ 2 replication.Material and Methods. The laboratory strain SARS‐CoV‐ 2/human/RUS/Nsk‐FRCFTM‐1/2020 was passivated on Vero cell culture. For the preparation of water extracts six types of experimental tea compositions (not containing flavorings) based on black and green tea or flowers of hibiscus as well as individual ingredients were used. Antiviral (inhibitory) activity of plant raw materials was studied in vitro according to the classical scheme of neutralization (inactivation) of the virus.Results. When compared with the main control sample of an aqueous extract of chaga (Inonotus obliquus) with a 50% effective concentration equal to 13.72±2.99 μg/ml against 103 TCPD50/ml of SARS‐CoV‐2, superior activity was found for extracts of the spice Syzygium aromaticum L. and black tea from Nepal. Approximately equal inhibitory activity was detected for extracts of tea compositions based on black tea with the addition of the grass Thymus altaicus and the spice S. aromaticum L. or with the addition of seeds of Carum carvi L. and the root of Angelica archangelica L. In addition, such activity was detected for extracts of tea compositions based on green tea from China with the addition of leaves of Mentha piperita L. and flowers of Lavandula angustifolia MILL. or with the addition of the peel of Citrus sinensis L. and seeds of Pimpinella anisum L. Extracts from individual plant raw materials used to make tea compositions were black teas from India, Argentina, Vietnam and Sri Lanka, leaves of M. piperita L. and flowers of L. angustifolia MILL. Cetraria islandica lichen, green tea and berries of Oxycoccus also showed antiviral activity.Conclusion. The results obtained suggest that tea compositions based on black and green tea with the addition of various plant raw materials in the form of a regular drink can be useful to people with COVID‐19 infection.

The Anti-Inflammatory Properties of Chaga Extracts Obtained by Different Extraction Methods against LPS-Induced RAW 264.7.

Chaga, a sclerotia formed by the Inonotus obliquus fungus, has been widely recognized for a number of medicinal properties. Although numerous scientific investigations have been published describing various biological activities of chaga from different geographical locations, little work has focused on chaga harvested in the USA or extraction techniques to maximize anti-inflammatory properties. The aim of this study was to investigate the anti-inflammatory properties of chaga collected in Maine (USA) extracted using traditional aqueous (hot water steeping) methods against lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. Chaga extracts obtained from both conventional (ethanol/water) extraction methods and an accelerated solvent extraction method (ASE) at optimized conditions were compared to aqueous extracts (tea) obtained from chaga in the powder form (P) and powder form in tea bags (B) based on their effect on both nitric oxide (NO) production and pro-inflammatory cytokine expression, in particular, the expression of TNF-α, interleukin-6 (IL-6), and interleukin-β (IL-1β). Phenolic acid extracts from chaga and individual phenolic acid standards were also investigated for their effect on the same parameters. Results indicated that various chaga extracts have significant anti-inflammatory activity on LPS-stimulated RAW 264.7 cells. The inhibitory effect was through a decrease in the production of NO and the downregulation of TNF-α, IL-6, and IL-1β in RAW 264.7 macrophages. ASE1 (novel, optimized ethanol/water extraction) and P6 (six-minute steeping of powder in 100 °C water) extracts showed the highest inhibitory activity on NO production and on the expression of the inflammatory cytokines, compared to extracts obtained by conventional extraction methods.

A polysaccharide from Inonotus obliquus ameliorates intestinal barrier dysfunction in mice with type 2 diabetes mellitus

Type 2 diabetes mellitus is a global disease that endangers human health, and the need for the development of nontoxic treatment candidates is urgent. In the present work, one homogeneous polysaccharide from Inonotus obliquus (IN) was isolated, and the protective effect and mechanism of IN on type 2 diabetes mellitus were investigated from the aspects of the intestinal barrier. IN mainly consisted of 9 monosaccharides with a Mw of 373 kDa. IN attenuated body weight loss, alleviated pathological damage, and suppressed the production of proinflammatory cytokines. Additionally, IN repaired the intestinal barrier by upregulating the expression of Ki-67, ZO-1 and MUC2. Furthermore, the abundance of Firmicutes was significantly increased with IN treatment, while the levels of Bacteroidetes were significantly inhibited. In conclusion, IN protected against type 2 diabetes mellitus by ameliorating intestinal barrier dysfunction and might serve as a novel drug candidate for type 2 diabetes mellitus.

Ziołolecznictwo w infekcjach wirusowych

The study is a review of natural raw materials that can prevent infection and help treat viral infections, including those that cause COVID-19. The condition of not getting infected with pathogens that cause infections of the upper and lower respiratory tract is high the body resistance. An important element that influences the proper immunity of the body is the diet. The functioning of the immune system is improved by bee products, and plant materials: purple coneflower herb, flower and root, licorice root, aloe gel and Baikal skullcap root, as well as black cumin seed oil, chaga mushroom, lemon balm leaves and chamomile flowers. Strengthening immunity is conducive to maintaining a good mood and reducing stress. The antiviral activity has been confirmed for many plant materials, especially those containing essential oils. Natural products can be used for prevention and treatment. The country that copes best with the coronavirus epidemic is China, thanks to a combination of academic and natural medicine. The study quotes an excerpt from the “Handbook of COVID-19 Prevention and Treatment”, prepared by Chinese doctors, with particular attention to the recipes used by them.

Study of the sterile conk of Inonotus obliquus using 13C CPMAS NMR and FTIR spectroscopies coupled with multivariate analysis

The sterile conk caused by the infection of the basidiomycete Inonotus obliquus (Chaga) is an important source of bioactive compounds. However, its structure and biochemistry are only generally understood. Solid state 13C NMR and FTIR spectroscopy have been utilized for the first time to investigate the sterile conk with non-invasive methods. The application of multivariate data analysis techniques and spectral distance algorithm to the obtained datasets showed clear distinction between the outer and inner layers of the sterile conk. Moreover, the sterile conk bark, compared to the inner layers, was spectroscopically more similar to wood tissues. The fungal tissue was proven to be concentrated below the bark. The similarity of the sterile conk inner layers to both decayed wood and hyphae of I. obliquus was shown by the multivariate data analysis of both spectra datasets. The spectroscopic data indicated lack of lignin degradation in the heart rot, except for demethoxylation, and a slight preference for hemicellulose degradation. Therefore, the results obtained suggest that the classification of I. obliquus as preferential lignin degrader (white-rot fungus) should be revised and clarified by further studies.

Inonotsuoxide B suppresses hepatic stellate cell activation and proliferation via the PI3K/AKT and ERK1/2 pathway

Hepatic stellate cells (HSCs) serve a pivotal role in the formation and degradation of the extracellular matrix during liver fibrosis. Inonotsuoxide B is a tetracyclic triterpenoid that can be extracted from Inonotus obliquus and has been previously reported to inhibit the growth of liver and gastric cancer cells. However, its effect on liver fibrosis remain poorly understood. Therefore, in the present study, the potential antiproliferative effects of inonotsuoxide B on HSCs was investigated. Initially, cells were divided into the following five groups: Control; platelet-derived growth factor (PDGF)-BB (10 ng/ml); and PDGF-BB + inonotsuoxide B (5, 10 and 20 µg/ml) groups. Inonotsuoxide B treatment (5, 10 and 20 µg/ml) was revealed to reverse PDGF-BB-induced HSC proliferation. Furthermore, the protein expression of α-smooth-muscle actin (α-SMA) and type I collagen was significantly decreased in the inonotsuoxide B (10 and 20 µg/ml) groups compared with the PDGF-BB group. Inonotsuoxide B (5, 10 and 20 µg/ml) was also revealed to suppress PDGF-BB-induced α-SMA mRNA expression and activation of the PI3K/AKT and ERK signaling pathways in HSCs. These findings suggest that inonotsuoxide B suppresses the proliferation and activation of HSCs by inhibiting the PI3K/AKT and ERK1/2 signaling pathways.

In Vitro Immunomodulatory Effects of Inonotus obliquus Extracts on Resting M0 Macrophages and LPS-Induced M1 Macrophages.

Background Inonotus obliquus (Chaga) is a parasitic fungus that is distributed mainly in northeast China. Our literature research showed chaga polysaccharides have bilateral effects on tumor necrosis factor (TNF)-α and interleukin (IL)-1β levels when they exert antitumor and antidiabetic activities. The current research tried to explore the influence of chaga extracts on inflammatory factors via macrophage polarization which has bilateral immune-regulation not only on healthy tissue homeostasis but also on pathologies. Methods Chaga was extracted with 100°C water and precipitated with 80% ethanol. The extracts were studied on RAW264.7 macrophage at resting condition (M0) and lipopolysaccharide (LPS)-activated subtype (classic activated macrophage, M1). The IL-1β, TNF-α, nitric oxide (NO) level, and the protein expressions of M1 and alternative activated macrophage (M2) markers including IL-1β, inducible NO synthase (iNOS), mannose receptor (CD206), and arginase (Arg)-1 were compared. Results The 100 g extracts contained 13.7 g polysaccharides and 1.9 g polyphenols. Compared with M0, the 50 μg/mL extracts increased NO level (P < 0.05) and decreased CD206 and Arg-1 expression significantly (P < 0.05). The extracts at 100–200 μg/mL increased NO and TNF-α level (P < 0.05), but increased iNOS and IL-1β expression significantly (P < 0.05). Compared with M1, the extracts decreased NO level at 25, 50, 100, and 200 μg/mL and decreased IL-1β and TNF-α level at 100–200 μg/mL significantly (P < 0.05). At 25–200 μg/mL, the extracts significantly increased CD206 and Arg-1 expression and decreased IL-1β and iNOS expression separately (P < 0.05). Conclusions Our research suggested that the bilateral effects of the chaga extracts on iNOS, IL-1β, and NO level on M0/M1 macrophages might be related with chaga polysaccharides and chaga polyphenols. Some in vivo anticancer and antidiabetic research of purified chaga polysaccharides related to macrophage differentiation should be conducted further.

Triterpenoids as bivalent and dual inhibitors of acetylcholinesterase/butyrylcholinesterase from the fruiting bodies of Inonotus obliquus

Inonotus obliquus, an edible and medicinal mushroom parasitic on birches, has been used in human diet and for traditional therapies in the high latitude regions of Europe and Asia for a long time. Our phytochemical study of this fungus led to the identification of fourteen triterpenoids including four undescribed ones, and two pairs of undescribed phenolic enantiomers. The undescribed compounds were elucidated by extensive spectroscopic analysis including 1D and 2D NMR and HRESIMS, quantum chemical NMR and ECD calculations, as well as single-crystal X-ray diffraction analysis. Bioassays revealed that eight compounds showed dual inhibition against acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) with IC50 values ranging from 2.40 ± 0.05 to 28.72 ± 0.46 μM, while 3β-hydroxy-lanosra-8,24-dien-21-al and trametenolic acid only presented BuChE inhibitory activities with IC50 values of 22.21 ± 1.01 and 7.68 ± 0.13 μM, respectively. In the kinetic studies, the most active three compounds acted as non-competitive inhibitors for both cholinesterases. Furthermore, molecular docking simulations revealed that three compounds demonstrated dual-sites bounding to AChE/BuChE. These triterpenoids emerged as bivalent and dual inhibitors of AChE/BuChE and could be effective drug candidates to prevent and treat Alzheimer's disease in the future.

Interference of Chaga mushroom terpenoids with the attachment of SARS-CoV-2; in silico perspective

Finding a potent inhibitor to the pandemic SARS-CoV-2 is indispensable nowadays. Currently, in-silico methods work as expeditious investigators to screen drugs for possible repurposing or design new ones. Targeting one of the possible SARS-CoV-2 attachment and entry receptors, Glucose-regulated protein 78 (GRP78), is an approach of major interest. Recently, GRP78 was reported as a recognized representative in recognition of the latest variants of SARS-CoV-2. In this work, molecular docking and molecular dynamics simulations were performed on the host cell receptor GRP78. With its many terpenoid compounds, Chaga mushroom was tested as a potential therapeutic against the SARS-CoV-2 receptor, GRP78. Results revealed low binding energies (high affinities) toward the GRP78 substrate-binding domain β (SBDβ) of Chaga mushroom terpenoids. Even the highly specific cyclic peptide Pep42, which selectively targeted GRP78 over cancer cells in vivo, showed lower binding affinity against GRP78 SBDβ compared to the binding affinities of terpenoids. These are auspicious results that need to be tested experimentally. Intriguingly, terpenoids work as a double sword as they can be used to interfere with VUI 202,012/01, 501.V2, and B.1.1.248 variants of SARS-CoV-2 spike recognition.

Anti-malarial activity in a Chinese herbal supplement containing Inonotus obliquus and Panax notoginseng.

Plasmodium falciparum, the most virulent human malaria parasite, causes serious diseases among the infected patients in the world and is particularly important in African regions. Although artemisinin combination therapy is recommended by the WHO for treatment of P. falciparum-malaria, the emergence of artemisinin-resistant parasites has become a serious issue which underscores the importance of sustained efforts to obtain novel chemotherapeutic agents against malaria. As a part of such efforts, thirty-nine herbal extracts from traditional Chinese medicine (TCM) were assayed for their anti-malarial activity using 3D7 strain of P. falciparum. Three herbal supplements appeared to possess higher specific anti-malarial activity than the others. One of them (D3) was separated by two sequential fractionations with reverse-phase (the first step) and normal-phase (the second step) liquid chromatography, in which some fractions resulted in higher specific activities than those of D3 or the previous fractions. Cell toxicity assay was performed with the fractions of the first fractionation and demonstrated no obvious cell toxicity. These results suggest that structure determination of the major compound for the anti-malarial activity in D3 may help the development of more potent chemicals in the future.

Qualitative analysis of secondary metabolites of chaga mushroom (Inonotus Obliquus): phenolics, fatty acids, and terpenoids

Chaga mushroom is a black perennial fungus that usually parasites on adult birch tree trunks. It has been conventionally used as a health-promoting supplement and nutraceutical in different cultures for centuries. The desired clarification of the profile of chaga secondary metabolites responsible for various bioactive properties has been continuously pursued for decades but has only partially been unveiled. Meanwhile, in recent years, attention to food safety, quality stability, authentication, and sustainability of chaga products from the wild has become increasingly popular in the current commercial market and related small/medium-size food industry enterprises. Phenolic, hydroxylated fatty acid, and terpenoid compounds produced by sclerotia of chaga mushrooms are bioactive constituents with antioxidant, anti-microorganism, and anti-tumor activities. Some new secondary metabolites of chaga mushroom have occasionally been reported previously, and effects of environment (e.g., cultivation method, harvesting region) on compositional characteristics noted. However, these have rarely and systematically compared the compositions of their material with a reliable database of known secondary metabolites of chaga. Therefore, this study aimed to achieve a rapid screening and characterization of secondary metabolites of Newfoundland chaga. A total of 111 phenolic, 63 fatty/aromatic acid, and 108 terpenoid constituents was primarily identified using HPLC-ToF-MS (high-performance liquid chromatograph coupled with time-of-flight mass spectra), among which 161 were newly reported. In addition, an update of the compositional database of chaga was provided as supplementary materials to help utilization and development of Newfoundland chaga mushroom as edible-fungi. Conclusively, chaga mushroom is a very promising food supplement abundant in numerous fungal secondary metabolites that were rarely found in other edible materials, even though its safety (e.g., oxalate content) aspects is still in need of additional investigation for being considered as a viable commercial nutraceutical.

Supercritical CO2 Extraction of Triterpenoids from Chaga Sterile Conk of Inonotus obliquus.

Triterpenoids are among the bioactive components of Chaga, the sterile conk of the medicinal fungus Inonotus obliquus. Supercritical fluid extraction of Chaga triterpenoids was carried out with supercritical CO2, while a modified Folch method was used as a comparison. Three temperature-pressure combinations were tested varying between 314–324 K (40–50 °C) and 281–350 bars, using time- and volume-limited extractions. Six triterpenoids were identified with GC-MS and quantified with GC-FID: ergosterol, lanosterol, β-sitosterol, stigmastanol, betulin, and inotodiol. The Folch extraction resulted in recovery of trametenolic acid, which was not extracted by supercritical CO2. Inotodiol was the major triterpenoid of all the extracts, with a yield of 87–101 mg/100 g and 139 mg/100 g, for SFEs and the Folch method, respectively. The contents of other major triterpenoids, lanosterol and ergosterol, varied in the ranges 59–63 mg/100 g and 17–18 mg/100 g by SFE, respectively. With the Folch method, the yields were 81 mg/100 g and 40 mg/100 g, respectively. The highest recovery of triterpenoids with SFE in relation to Folch was 56% and it was obtained at 324 K (50 °C) and 350 bar, regardless of extraction time or volume of CO2. The recoveries of lanosterol and stigmastanol were unaffected by SFE conditions. Despite the lower yield, SFE showed several advantages including shorter extraction time and less impact on the environment. This work could be a starting point for further studies on green extraction methods of bioactive triterpenoids from Chaga.

ELEMENTAL COMPOSITION OF CHAGA MUSHROOM (INONOTUS OBLIQUUS)

Birch chaga Inonotus obliquus is used as a component of many medicines. This is due to the versatile therapeutic effect associated with the content of a large number of biologically active substances of inorganic, organometallic and organic nature, a high content of macro- and microelements both in a free state and in the form of chelate complexes. To establish the qualitative and quantitative elemental composition of birch chaga, highly sensitive physicochemical methods of analysis were used, such as atomic emission spectroscopy (AES), inductively coupled plasma mass spectrometry (ICP-MS), and neutron activation analysis (NAA). The scientific novelty of this study lies in the conduct of a complex physicochemical elemental layer-by-layer analysis of chaga, the analogs of which have not been found in the available literature. The analyzed samples of birch chaga layers contain the following elements: K, Na, Mg, Ca, Zn, Mn, Cu, Fe, Mo, Co, Cr, P, Br, Se, V, Al, Ba, Li, Ag, Au, Rb, Cs, As, Sb, B, Sr, Pb, Ni, Ti, Be, Ta, U, Th, Sc; the concentration of elements is highest in the outer layer of the fungus. It is noted that the content of both biogenic elements (K, Co, Mn) and potentially toxic (Ag, Br, Rb) is significantly higher than the corresponding clarkes in bios. Differences in the content of elements correlate with modern biochemical concepts of the composition of chaga, and in most cases also correlate with the values of clarkes in bios. The value of the detected toxicant elements does not exceed the MPC.

Inonotus obliquus and its bioactive compounds alleviate non-alcoholic fatty liver disease via regulating FXR/SHP/SREBP-1c axis

Non-alcoholic fatty liver disease (NAFLD) is currently the most common chronic liver disease worldwide. However, there is still lack of specific drugs for treating NAFLD in clinic. Inonotus obliquus (IO), a folk medicinal fungus, has long been used to prevent against metabolic syndrome related diseases, such as hypertension and diabetes, etc. However, the study of IO anti-NAFLD effect has been reported rarely. This study aimed to investigate whether IO has an inhibitory effect on NAFLD, identify the active compounds in IO and clarify the underlying mechanisms of its anti-NAFLD effects. The results of Oil Red O(ORO) and Hematoxylin-Eosin (HE) staining, lipid extraction and determination showed that IO and its extracts, including inotodiol (Ino), lanosterol (Lan) and trametenolic acid (TA), could remarkably ameliorate lipid accumulation in MCD diet-induced mouse livers or OA-induced LO2 hepatocytes. Moreover, qPCR analysis revealed that IO and its compounds significantly downregulated the mRNA levels of lipogenic genes, such as SREBP-1c, ACC1 and FASN, and upregulated the mRNA levels of FXR and SHP. We found that the administration of guggulsterone (GS), a FXR inhibitor, abolished the inhibitory effect of Ino on lipid deposition in OA-induced LO2 cells. In conclusion, IO and its compounds attenuate hepatic lipid accumulation in NAFLD by inhibiting liver lipogenesis. The anti-NAFLD effects of Ino, a bioactive compound in IO, are through regulating FXR/SHP/SREBP-1c pathway. Our results suggested that IO and its bioactive compound Ino may become promising drugs to treat NAFLD.

Badania jakościowe i ilościowe wyciągów z Inonotus obliquus

Introduction. Inonotus obliquus (Ach. Ex Pers.) Pilát is a fungus found in North America, Finland, Poland, Russia, China, Japan and Korea on the bark of trees, most often Betula pendula, B. pubescens, B. carpatica and Fagus sylvatica. Mushroom extracts show a number of biological activities, due to the presence of: polysaccharides, phenolic compounds (phenolic acids and flavonoids), triterpenes, ergosterol and its peroxide. Aim. Determination of the content of the sum of polyphenolic compounds, including the sum of phenolic acids, flavonoids and flavonols in water, methanol and methanol-water extracts, determination of antioxidant activity and qualitative determination of the composition of the chloroform extract from the fruiting body of Inonotus obliquus, collected from a natural site. Material and methods. The raw material for research was collected in the Noteć Forest in the Sieraków Landscape Park. The fruiting body was dried and then ground into a fine powder. Using spectroscopic methods, the sum of polyphenols with the Folin-Ciocalteu reagent, the sum of phenolic acids using the Arnov method, the sum of flavonols with aluminum chloride, the sum of flavonoids according to the method described in FP X, antioxidant activity with the DPPH radical and the composition of the chloroform extract using GC-MS were determined. Results. The methanol extract was characterized by the highest content of the sum of phenolic acids per coffee acid (0.127%), and the lowest content of water-methanol extract (0.047%). The highest content of the sum of polyphenols in terms of caffeic acid was determined in the aqueous extract (1.85%), and the lowest in the methanol extract (1.01%). The methanol extract was characterized by the highest content of the sum of phenolic acids expressed as coffee acid (0.127%), and the lowest content of water-methanol extract (0.047%). The content of total flavonoids and total flavonols was determined only in methanol extracts, because the precipitation made it impossible to perform the determinations in the remaining extracts. The lowest IC50 value was determined for the water-methanol extract. GC-MS analysis allowed to identify, among others: isovanillin, santalene, α-selinene, syringaldehyde, linoleic acid, thymol, α-bergamotene and lanosterol. Conclusions. The highest content of the sum of polyphenols was determined in the water extract of Inonotus obliquus, the sum of phenolic acids in the methanol extract; the water-methanol extract was characterized by the strongest antioxidant properties. GC-MS analysis revealed the presence of numerous compounds with confirmed biological activity.

상업효소를 이용한 차가버섯 추출물의 생물전환 공정을 통한 생리활성물질 생산과 항산화 활성 증진

Background: Inonotus obliquus (Pers.:Fr.) Pilát is regarded as a functional food and medicinal resource due to the polyphenol content. However, the polyphenols in I. obliquus have a low absorption rate because they exist as a complex combined with proteins or carbohydrates. This study improves the production of free polyphenols through low molecularization in the bioconversion process using commercial enzymes including protease and carbohydrate-degrading enzymes.<BR>Methods and Results: The total polyphenol content and radical scavenging activity of I. obliquus extract was significantly enhacned after bioconversion by commercial enzymes when compared to a hot-water extraction (p < 0.05). In addition, the I. obliquus extract was isolated from the bioconverted product using high performance liquid chromatography to investigate the main substances in the bioconverted products. The main substance was syringic acid at a concentration 73.6% (using Protamex) and 60.4% (using Flavourzyme) higher than the hot-water extract (p < 0.05). <BR>Conclusions: The results confirm I. obliquus bioconverted using proteases contains a high anti-oxidant content when compared to the carbohydrate-degraded I. obliquus. Therefore, bioconverted I. obliquus using proteases should be developed as a high value-added raw material with excellent antioxidant potential for producing functional foods and medicines.

Isolation of chemical compositions as dietary antioxidant supplements and neuroprotectants from Chaga mushroom (Inonotus obliquus)

Excessive free radicals can cause oxidative stress (OS), which lead to neurodegenerative diseases (ND) are harmful to the human body. Therefore, it is necessary to explore dietary antioxidants and neuroprotectants from nutrient foods. Chaga mushroom (Inonotus obliquus) has been used as an important functional food in many countries for centuries. This present study was carried out to discover active compounds from I. obliquus and investigate the in vitro antioxidant activity and neuroprotective activity. Sixteen natural products including two new isocoumarins (1–2), a new 8-O-4′-neolignan (3), a new cyclic diarylheptanoid (4), and twelve known compounds (5–16) were separated from I. obliquus. Their structures were elucidated by extensive spectroscopic analyses of high resolution electrospray mass spectrometry (HRESIMS), ultra violet (UV), and nuclear magnetic resonance (NMR). Among them, compounds 1 and 2 revealed remarkable antioxidant activities through the comprehensive analysis of DPPH, ABTS, and ferric reducing antioxidant power (FRAP) methods. Meanwhile, compound 1 demonstrated significant neuroprotective activity by increasing SH-SY5Y cells viability and preventing mitochondrial damage. Apart from that, electronic analyses were performed using the highest occupied molecular orbital (HOMO), and the lowest unoccupied molecular orbital (LUMO) to analyze compound 1. These results indicated that compound 1 had a significant implication for the search of natural raw material to prevent OS and nerve damage in the field of functional foods, and the isocoumarin fragment as well as an oxygenated cis double-bond might be the active sites to play prominent parts in antioxidant and neuroprotective activities of compound 1.

Biological Synthesis of Bioactive Gold Nanoparticles from Inonotus obliquus for Dual Chemo-Photothermal Effects against Human Brain Cancer Cells.

The possibility for an ecologically friendly and simple production of gold nanoparticles (AuNPs) with Chaga mushroom (Inonotus obliquus) (Ch-AuNPs) is presented in this study. Chaga extract’s reducing potential was evaluated at varied concentrations and temperatures. The nanoparticles synthesized were all under 20 nm in size, as measured by TEM, which is a commendable result for a spontaneous synthesis method utilizing a biological source. The Ch-AuNPs showed anti-cancer chemotherapeutic effects on human brain cancer cells which is attributed to the biofunctionalization of the AuNPs with Chaga bioactive components during the synthesis process. Further, the photothermal ablation capability of the as-prepared gold nanoparticles on human brain cancer cells was investigated. It was found that the NIR-laser induced thermal ablation of cancer cells was effective in eliminating over 80% of the cells. This research projects the Ch-AuNPs as promising, dual modal (chemo-photothermal) therapeutic candidates for anti-cancer applications.

Extraction possibilities of lipid fraction and authentication assessment of chaga (Inonotus obliquus)

Chaga (Inonotus obliquus) (Fr.) Pilát is a black perennial fungus that grows on adult birch trunks and has been traditionally used as a health promoting remedy in different countries. The lipophilic compounds of chaga were isolated applying conventional (Soxhlet), intensive (ultrasound assisted, accelerated solvent extraction) and environmentally friendly (supercritical fluid) extraction methods utilizing both polar and non-polar solvents. Soxhlet extraction showed the highest inotodiol yield (231 mg/100 g chaga) using cyclohexane. For authentication of chaga samples, isotope ratio mass spectrometry (IRMS) and multi-elemental analysis was performed. The extraction yield and profile of lipids, sterols and triterpenoids depend on the chaga origin. The amount of lipids in chaga samples varied in the range from 0.5 to 1.0%. The antimicrobial activity of chaga extracts against pathogenic and opportunistic pathogenic microorganisms was demonstrated. The results showed that extraction using supercritical CO2 delivered yields comparable to those obtained by conventional and intensive extraction techniques. Amount of target compounds (sterols and triterpenoids) obtained using supercritical CO2 extraction could represent the future for sustainable industrial-scale production of biologically active ingredients with antimicrobial activity. Authentication using IRMS allowed to differentiate between natural (fruiting bodies) and cultivated (mycelium) chaga.

Renal Protective Effects of Inonotus obliquus on High-Fat Diet/Streptozotocin-Induced Diabetic Kidney Disease Rats: Biochemical, Color Doppler Ultrasound and Histopathological Evidence.

Diabetic kidney disease (DKD) is the current leading cause of end-stage renal disease. Inonotus obliquus (chaga), a medicinal fungus, has been used in treatment of diabetes. Here, we aim to identify the renal protective effects of chaga extracts on a DKD rat model which was induced by a high-fat diet and streptozotocin injection. During the total 17-weeks experiment, the biological parameters of serum and urine were examined, and the color Doppler ultrasound of renal artery, the periodic acid-Schiff staining, and electron microscopy of kidney tissue were performed. The compositions of chaga extracts were analyzed and the intervention effects of the extracts were also observed. Compared with the normal control group, the biochemical research showed that insulin resistance was developed, blood glucose and total cholesterol were elevated, urinary protein excretion and serum creatinine levels were significantly increased in the DKD model. The ultrasound examinations confirmed the deteriorated blood flow parameters of the left renal interlobar artery in the rat models. Finally, histopathological data supported renal injury on the thickened glomerular basement membrane and fusion of the foot processes. 8 weeks intervention of chaga improved the above changes significantly, and the 100 mg/kg/d chaga group experienced significant effects compared with the 50 mg/kg/d in some parameters. Our findings suggested that Doppler ultrasound examinations guided with biochemical indicators played important roles in evaluating the renal injury as an effective, noninvasive, and repeatable method in rats. Based on biochemical, ultrasound, and histopathological evidence, we confirmed that chaga had pharmacodynamic effects on diabetes-induced kidney injury and the aforementioned effects may be related to delaying the progression of DKD.