Chaga mushroom (Inonotus obliquus) induces G0/G1 arrest and apoptosis in human hepatoma HepG2 cells

AIM: To investigate the anti-proliferative and apoptotic effects of Chaga mushroom (Inonotus obliquus) water extract on human hepatoma cell lines,HepG2 and Hep3B cells. METHODS: The cytotoxicity of Chaga extract was screened by 3-4,5-dimethylthiazol-2-yl-2,5-diphenyltetrazolium bromide (MTT) assay. Morphological observation,flow cytometry analysis,Western blot were employed to elucidate the cytotoxic mechanism of Chaga extract. RESULTS: HepG2 cells were more sensitive to Chaga extract than Hep3B cells,as demonstrated by markedly reduced cell viability. Chaga extract inhibited the cell growth in a dose-dependent manner,which was accompanied with G0/G1-phase arrest and apoptotic cell death. In addition,G0/G1 arrest in the cell cycle was closely associated with down-regulation of p53,pRb,p27,cyclins D1,D2,E,cyclin-dependent kinase (Cdk) 2,Cdk4,and Cdk6 expression. CONCLUSION: Chaga mushroom may provide a new therapeutic option,as a potential anticancer agent,in the treatment of hepatoma.

Inonotus obliquus(Chaga)against HFD/STZ-induced glucolipid metabolism disorders and abnormal renal functions by regulating NOS-cGMP-PDE5 signaling pathway

Our prior investigations have established that Inonotus obliquus(Chaga)possesses hypoglycemic effects.Persistent hyperglycemia is known to precipitate renal function abnormalities.The functionality of the kidneys is intricately linked to the levels of cyclic guanosine-3',5'-monophosphate(cGMP),which are influenced by the activities of nitric oxide synthase(NOS)and phosphodiesterase(PDE).Enhanced cGMP levels can be achieved either through the upregulation of NOS activity or the downregulation of PDE activity.The objective of the current study is to elucidate the effects of Chaga on disorders of glucolipid metabolism and renal abnormalities in rats with type 2 diabetes mellitus(T2DM),while concurrently examining the NOS-cGMP-PDE5 signaling pathway.A model of T2DM was developed in rats using a high-fat diet(HFD)combined with streptozotocin(STZ)administration,followed by treatment with Chaga extracts at doses of 50 and 100 mg·kg^(−1)for eight weeks.The findings revealed that Chaga not only mitigated metabolic dysfunctions,evidenced by improvements in fasting blood glucose,total cholesterol,triglycerides,and insulin resistance,but also ameliorated renal function markers,including serum creatinine,urine creatinine(UCr),blood urea nitrogen,24-h urinary protein,and estimated creatinine clearance.Additionally,enhancements in glomerular volume,GBM thickness,podocyte foot process width(FPW),and the mRNA and protein expressions of podocyte markers,such as nephrin and wilms tumor-1,were observed.Chaga was found to elevate cGMP levels in both serum and kidney tissues by increasing mRNA and protein expressions of renal endothelial NOS and neural NOS,while simultaneously reducing the expressions of renal inducible NOS and PDE5.In summary,Chaga counteracts HFD/STZ-induced glucolipid metabolism and renal function disturbances by modulating the NOS-cGMP-PDE5 signaling pathway.This research supports the potential application of Chaga in the clinical prevention and treatment of T2DM and diabetic nephropathy(DN),with cGMP serving as a potential therapeutic target.

Effects of Inotodiol Extracts from Inonotus Obliquus on Proliferation Cycle and Apoptotic Gene of Human Lung Adenocarcinoma Cell Line A549

Objective： To observe the proliferation inhibition, apoptosis, and cell proliferation cycle of human lung carcinoma cell line A549 treated with Inotodiol extracts from Inonotus obliquus and explore the possibility of Inotodiol extracts from Inonotus obliquus as a new tumor chemopreventive drug. Methods： Human lung cancer cell line A549 was treated with different concentrations of Inotodiol, the effects of Inotodiol on cell apoptosis, the expression of Ki-67, Bcl-2, Bax, and p53 and cell cycle were detected by TUNEL assay, immunohistochemistry, and flow cytometry assay respectively. Results： Inotodiol extracts had antiproliferation effect on human lung carcinoma cell line A549. The expression of Ki-67 decreased with the increase of Inotodiol concentration and exposure time （P〈0.05）, in a close-dependent and time-dependent manner. The typical characteristics of the apoptosis of A549 cells treated with Inotodiol were observed, and the apoptotic rate of A549 cell at 48 h was the highest by TUNEL assay. Inotodiol arrested A549 cells in the S phase, and apoptotic peak was observed by flow cytometry. Immunocytochemistry indicated that the expression of Bcl-2 protein decreased, while the expression of p53 and Bax proteins increased in A549 cells treated with Inotodiol, compared with the control cells （P〈0.05）. Conclusion： Inotodiol can inhibit proliferation and induce the apoptosis of A549 cells, and its molecular mechanism may be associated with the up-regulating expression of p53 and bax proteins and down- regulating expression of Bcl-2 protein, which arrested A549 cells in S phase.

Progress of Research on Inonotus Obliquus

Inonotus obliquus has high nutritional and medicinal value,especially in treating malignant tumors,diabetes,cardiovascular disease and AIDS,attracting significant attention from scholars in recent years.In this paper,the biological characteristics,chemical composition and pharmacologic effects of Inonotus obliquus were summarized.And the applications in medicine and food were introduced.Future research on Inonotus obliquus was also discussed in order to make Inonotus obliquus obtain effective exploitation a...

Aqueous Two-phase Systems with Ultrasonic Extraction Used for Extracting Phenolic Compounds from Inonotus obliquus

Objective To optimize the extracting technology of assessing the maximum yield of phenolic compounds (PC) from Inonotus obliquus by single factor experiments and orthogonal array design methods through aqueous two-phase systems combined with ultrasonic extraction. Methods The range of the independent variables, namely levels of acetone and ammonium sulfate, and ultrasonic time were identified by a first set of single factor experiments. The actual values of the independent variables coded at four levels and three factors were selected based on the results of the single factor experiments. Subsequently, the levels of acetone and ammonium sulfate, and ultrasonic time were optimized using the orthogonal array method. Results The optimum conditions for the extraction of PC were found to use 7.0 mL acetone, 5.5 mg ammonium sulfate, with ultrasonic time for 5 min. Under these optimized conditions, the experimental maximum yield of PC was 37.8 mg/g, much higher than that of the traditional ultrasonic extraction (UE, 29.0 mg/g). And the PC obtained by this method had stronger anti-oxidative activities than those by traditional UE method. Conclusion These results indicate the suitability of the models developed and the success in optimizing the extraction conditions. This is an economical and efficient method for extracting polyphenols from I. obliquus.

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 Activity of Fomes fomentarius

In this work the antioxidant and antimicrobial activity of the fractions with different polarity,obtained from water-ethanol dry extracts from Fomes fomentarius,has been investigated.Fractions of different polarities,obtained from water-alcohol dry extract from Inonotus obliquus,were used as comparison samples.It has been shown,that the most phenol-rich(73.571±21.268 mg E/g)ethyl acetate and ethanol(36.343±10.241 mg E/g)fractions from Fomes fomentarius demonstrated the highest radical scavenging activity against DPPH(IС50:515.6±100.5μg/ml;751.2±19.3μg/ml,respectively)and ABTS(IС_(50):229.7±9.6μg/ml;423.3±0.66μg/ml,respectively).It has been found,that chloroform,ethyl acetate and ethanol fractions from Fomes fomentarius exhibit antimicrobial activity against Staphylococcus aureus.