Establishment of TLC identification and HPLC fingerprint of Inonotus obliquus and analysis of chemical pattern recognition / 中国药房

OBJECTIVE To establish thin-layer chromatography （TLC） identification method and high-performance liquid chromatography （HPLC） fingerprint of Inonotus obliquus， and to evaluate the quality of I. obliquus by chemical pattern recognition. METHODS TLC method was used to identify trametenolic acid and inotodiol in I. obliquus qualitatively. HPLC fingerprint of I. obliquus was established； Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine （2012 edition） was used to determine the common peaks and evaluate the similarity； chemical pattern recognition analysis [cluster analysis， principal component analysis and orthogonal partial least squares-discriminant analysis （OPLS-DA）] of 22 batches of I. obliquus was performed with SPSS 23.0 software and SIMCA14.1 software. RESULTS In the TLC， the same color spots were found at the same position in the chromatograms of test sample and substance control. A total of 10 common peaks were marked in the HPLC fingerprints of 22 batches of I. obliquus， with similarities of 0.942-0.995. No. 3 peak was identified as trametenolic acid， No.4 peak as inotodiol， No. 9 peak as ergosterol and No. 10 peak as lanosterol. Results of cluster analysis showed that S1-S15， S19， S21 and S22 could be clustered into the first category， and S16-S18 and S20 were clustered into the second category. Results of principal component analysis showed that top 4 samples in the list of comprehensive score were S17， S18， S16 and S20. Results of OPLS-DA showed that three marking components that may affect the quality of I. obliquus were screened according to the standard of VIP＞1， i.e. No. 4 peak （inotodiol， VIP value of 1.86）， No. 3 peak （trametenolic acid， VIP value of 1.62） and No. 7 peak （VIP value of 1.27）. CONCLUSIONS This study establishes TLC method and HPLC fingerprint of I. obliquus successfully， which can provide reference for the quality control of I.obliquus by combining with chemical pattern recognition.

Study on the purification technology of total triterpenoid extracts from Inonotus obliquus and their anti -tumor activity in vitro / 中国药房

OBJECTIVE To optimite the purification technology of total triterpenoid extracts from Inonotus obliquus ，and to investigate the anti -tumor activity of its purified products . METHODS Using inotodiol as control ，the method was established for the content determination of total triterpenoid in I. obliquus. The type of macroporous adsorption resin ，sample volume ，sample concentration，sample flow rate ，eluent volume ，eluent dosage and elution flow rate were selected by single factor experiments . The purification technology of the crude extract was determined and verified . The effects of total triterpenoid purified from I. obliquus on the proliferation ，migration and apoptosis of human cervical cancer HeLa cells were detected by cell proliferation test ， migration test ，flow cytometry and AO/EB kit . RESULTS The best purification technology of total triterpenoid crude extracts from I. obliquus was as follows ：AB-8 macroporous adsorption resin was used ；mass concentration of the sample solution was 2.0 mg/mL；sample volume was 140 mL，and the flow rate was 1.0 mL/min；the impurity was removed with 50% ethanol 40 mL， then eluted with 95% ethanol 160 mL，at the elution flow rate of 3.0 mL/min. After purification ，mass concentration of total triterpenoid from I. obliquus increased from 34.36% to 73.39%. The total triterpenoid of I. obliquus could inhibit the proliferation of HeLa cells ，and the 50% inhibitory concentration was 184.20 μg/mL. Compared with control group ，the purified products could significantly inhibit the migratio n and promote the apoptosis of HeLa cells （P＜0.05 or P＜0.01）. CONCLUSIONS The purification technology of total triterpenoids extracts from I. obliquus is successfully optimited . The purified product could inhibit the proliferation and migration of HeLa cells and induce their apoptosis.

Analysis on the compatible application of inonotus obliquus in the treatment of hepatobiliary disorders / 国际中医中药杂志

According to the relevant data of Russia, this paper summarized and analyzed the compatibility and application methods of Inonotusobliguus in the rehabilitation process of treating hepatobiliary disorders. In the rehabilitation process of a variety of hepatobiliary disorders, Inonotusobliguus can be used with Berberis vulgaris L., Achillea millefolium L., Hypericum perforatum L., Acorus calamus L., Calendula officinalis L., Cichoriumintybus L., Urticadioica L., Artemisia draculoides, Tanacetum vulgare L., Taraxacum officinale Wigg., Agropyronrepens (L.) Pal.Beauv, Rosa canina L., Carumcarvi L. and Majorana hortensis Moench. and other compatible applications. They have certain effect on jaundice, hepatitis, chronic cholecystitis, and gallbladder pain.

Studies on the anti-tumor efficacy of inonotus obliquus in Russia: A literature review / 国际中医中药杂志

In this paper, we reviewed the findings from previous studies on the anti-tumor efficacy of inonotus obliquus, one of the most famous edible and medical fungi, in Russia and the former Soviet Union since the 1950s. To be specific, the findings from clinical researches regarding the medicinal benefits of inonotus obliquus for treating gastric cancer, esophageal cancer, lung cancer, laryngeal cancer, breast cancer, and leukemia have been summarized, indicating inonotus obliquus may relieve symptoms, improve immune and neural function, improve life quality, and may prolong survival in the cancer patients.

Inonotus obliguus as medical fungi in Russia / 国际中医中药杂志

This paper reviewed characteristics of inonotus obliquus, and progress in medical research regarding inonotus obliquus in Russia. To be specific, we summarized morphological characteristics, distribution, growing conditions, history of application, active ingredients, and mechanism of actions of inonotus obliquus in this review, and illustrated specific notes for its potential medicinal value and health benefits. Our manuscript has provided new insight for further research and application of inonotus obliquus in China.

Further analysis of the compatibility application of Inonotus obliquus in the treatment of gastrointestinal disordersin Russia / 国际中医中药杂志

This paper introduces the application of Inonotus obliquus in the rehabilitation treatment of gastrointestinal diseases in Russia. It is found that Inonotus obliquus is widely used in the treatment of gastrointestinal diseases. Inonotus obliquus can be compatible with Mentha nigrum, green walnut peel, Hypericum perforatum, chamomile, red Sorbus, Daisy, yarrow, Schisandra, Tilia, chamomile, fennel seed, marigold, plantain juice, dried potted leaves, burdock root, etc., and it has a good effect on diarrhea, constipation, gastric and duodenal ulcer, low acid gastritis, acute gastritis, dodecadactyly Enteritis and so on have certain curative effect.

Analysis on the compatibility of Inonotus obliquus in the treatment of gastrointestinal disordersin Russia / 国际中医中药杂志

Inonotus obliquus is a commonly used edible fungus in Russia. For centuries, it has been used to treat digestive tract diseases and tumors in the northern Russian countryside. Based on the relevant researches of the Soviet Union and Russia, this paper summarizes and analyzes the compatibility application of inonotus obliquus in the rehabilitation of gastrointestinal diseases. In the rehabilitation treatment of various gastrointestinal diseases, inonotus obliquus can be used alone or in combination with other drugs. In China, the difference in the source and the dosages should be paid the attention before the application..

Analysis on the compatibility of Inonotus obliquus in the treatment of tumor in Russia / 国际中医中药杂志

Inonotus obliquus is a commonly used medicinal and edible fungus in Russian folk. It is often used in the rehabilitation treatment of cancer patients. Based on the relevant research data of the Soviet Union and Russia, this paper summarizes and analyses the compatibility application methods of Inonotus obliquus in cancer rehabilitation treatment. It is believed that Inonotus obliquus can be used alone as tea or in the combination with other drugs. The prescription is simple, practical, safe and easy for patients, but it should be tested in our country. Considering the different constitution of patients and the source of medicinal materials, the attention should be paid to adjust the dosage.

Analysis on the compatibility of Inonotus obliquus in the treatment of gastrointestinal disordersin Russia / 国际中医中药杂志

Inonotus obliquus is a commonly used edible fungus in Russia. For centuries, it has been used to treat digestive tract diseases and tumors in the northern Russian countryside. Based on the relevant researches of the Soviet Union and Russia, this paper summarizes and analyzes the compatibility application of inonotus obliquus in the rehabilitation of gastrointestinal diseases. In the rehabilitation treatment of various gastrointestinal diseases, inonotus obliquus can be used alone or in combination with other drugs. In China, the difference in the source and the dosages should be paid the attention before the application..

Analysis on the compatibility of Inonotus obliquus in the treatment of tumor in Russia / 国际中医中药杂志

Inonotus obliquus is a commonly used medicinal and edible fungus in Russian folk. It is often used in the rehabilitation treatment of cancer patients. Based on the relevant research data of the Soviet Union and Russia, this paper summarizes and analyses the compatibility application methods of Inonotus obliquus in cancer rehabilitation treatment. It is believed that Inonotus obliquus can be used alone as tea or in the combination with other drugs. The prescription is simple, practical, safe and easy for patients, but it should be tested in our country. Considering the different constitution of patients and the source of medicinal materials, the attention should be paid to adjust the dosage.

Characteristics of potential effects and benefits of inonotus obliquus from the perspective of traditional Chinese medicine / 国际中医中药杂志

According to the literatures pertaining inonotus obliquus (commonly known as chaga) documented in Russia and the former Soviet Union, the research progress of inonotus obliquus since the 1950s in Russia and the former Soviet Union and latest research progress made over the last two decades in China, , the authors further elaborated potential effects and benefits of inonotus obliquus from the perspective of traditional Chinese medicine (TCM) based on prior understanding of Flavor Effect of Inonotus obliquus. It is believed that Inonotus obliquus can benefit the five internal organs, invigorating the circulation of qi, blood, and nutriment, and thus may improve the body's adaptability and self-protection when exposing to pathogenic factors. The mechanisms of the multiple effects of Inonotus obliquus have the characteristics of multi-component, multi-target, multi-system, and multi-pathway. Inonotus obliquus can be considered to be used as both TCM medication and TCM dietary supplement.

Understanding of traditonal Chinese medicine flavor effect of inonotus obliquus / 国际中医中药杂志

According to the literatures of the former Soviet Union and Russia about inonotus obliquus (commonly known as chaga),and the research progress in the past 20 years in China,and the author's 30-year experience on inonotus obliquus' clinical application,and the records of medicinal properties of birch in ancient China literatures,the potential effect of inonotus obliquus is analyzed according to traditional Chinese medicine theory.It is believed that the taste of inonotus obliquus is slightly bitter and cold.The main effect of inonotus obliquus is strengthening the body resistance,the specific benefits of which include nourishing qi,nourishing blood,nourishingyin,tonifying spleen,invigorating the spleen and stomach,nourishing liver,tonifying kidney and nourish the heart and calm the nerves.The effect of inonotus obliquus also includes eliminate evil (pathogenic factors),with clearing heat and detoxicating,soothing liver and relieving stagnation,and promoting blood circulation and resolving hard lump.

Studies on the anti-tumor efficacy of inonotus obliquus in Russia: A literature review / 国际中医中药杂志

This paper summarized the anti-tumor experimental researches of inonotus obliquus in Russia and former Soviet Union.Since the 1950s,a series of experimental studies of Inonotus obliquus have been carried out in Russia and the Soviet Union based on prior clinical studies to further explore its anti-tumor effects and mechanisms.The effects of Inonotus obliquus were inhibiting the growth and metastasis of tumors,and its specific mechanisms might be involving regulation of cancer cell dynamic phenotypes.

A study on apoptosis of fermented fnonotus obliquus on hepatocellular carcinoma HepG2 cells / 医学研究生学报

Objective A variety of secondary metabolites can be obtained after the fermentation of medicinal fungi.The con-tent of these metabolites is much higher than that of cultivated medicinal fungi. We aimed to preliminarily study on the apoptosis of fer-mented Inonotus obliquus(FIO) on hepatocellular carcinoma HepG-2 cells. Methods The experiment was divided into 3 groups:the control group(The same volume of drug-free medium),the UIO group(200 mg/L) and the FIO group(200 mg/L). The contents of polysaccharides,terpenoids and polyphenols in unfermented Inonotus Obliquus (UIO) and FIO were determined by sulfuric acid phenol method,Folin-Ciocalteu method and colorimetric method. Effects of Fermented Inonotus obliquus on apoptosis of HepG-2 Cells were measured by MTT Method and Flow Cytometry. Results The contents of polysaccharides,terpenoids and polyphenols were sig-nificantly increased in FIO group [(10.140±0.849)mg/mL,(1.774±0.001)mg/mL, (9.979±0.022)mg/mL] compared with thatin UIO group [(7.161±0.305)mg/mL,(1.358±0.004)mg/mL,(6.314±0.237)mg/mL](P<0.01). Both UIO and FIO group induced apoptosis of HepG-2 cells according to Annexin V/PI double staining. Compared with the control group(4.3%),the apoptosis rate in-creased in UIO and FIO group (23.14%,27.37%) (P<0.05). In addition, the apoptosis rate was higher in FIO group than that in UIO group (P<0.05). Compared with the control group, the ratio of G0/G1phase cells was significantly increased and the ratio of S phase and G2/M phase was significantly decreased in UIO and FIO group (P<0.01). G0/G1phase cell ratio differences were statistically signif-icant compared with the UIO group (P<0.05) Conclusion FIO can better induce the apoptosis of HepG-2 cells than UIO.

Research progress of inonotus obliquus in anti tumor / 医学研究生学报

Chemical synthetic drugs are the main therapeutic drugs for cancer, however, the time-consuming drug development, lengthy treatment, serious side effects and drug resistant of chemical synthetic drugs significantly increase the burden of the patients.In recent years, natural anti tumor substances with good treatment effectiveness and low toxicity, have attracted more and more attentions in studying anti tumor drugs.It is reported that Inonotus obliquus can significantly suppress tumors, while the antitumor components and mechanisms of Inonotus obliquus are rarely reported.In this paper, we will review the antitumor effect, antitumor components and mechanisms of Inonotus obliquus.

Simultaneous determination of seven nucleosides in inonotus obliquus by HPCE / 中国药学杂志

OBJECTIVE: To establish a high performance capillary electrophoresis (HPCE) method for simultaneous determination of seven nucleosides and nucleobases, including adenine, uracil, adenosine, hypoxanthine, uridine, guanosine, and inosine in Inonotus obliquus samples of different batches. METHODS: Based on the mode of HPCE, uncoated fused silica capillary (75 μm × 64.5 cm, 56 cm of effective length) was used with separation voltage of 20 kV. The 55 mmol·L-1 sodium borate solution was selected as the running buffer solution (pH 9.5). The detection wavelength was set at 260 nm. The sample was injected at 5 kPa × 6 s and column temperature was maintained at 30℃. RESULTS: Good linear relationship was obtained between the response peak areas and the concentrations of all the seven nucleosides (r > 0.998 8). The average recoveries of the method were between 96.46%-102.12%. The contents of the seven nucleosides in the Inonotus obliquus samples of different origins were different. CONCLUSION: The established method is reliable, accurate, and can be used for the quality control of Inonotus obliquus.

The research progress in antitumor mechansim of Inonotus obliquus / 实用肿瘤学杂志

Since the tumor has become one of the most deadly diseases to human life ,it is of great impor-tance to explore a new effective antitumor medicine .This paper summarizes the chemical constituents and the mechanism of Inonotus obliquus .The mechanism includes the details of the inhibition of tumor cell proliferation , inducing tumor cell apoptosis ,influence tumor cell cycle etc .Meanwhile,it describes Inonotus obliquus to the ad-juvant chemotherapy of tumor .

Induction of human MCF-7 breast cancer cell apoptosis by Inonotus obliquus and study on its mechanism / 中草药

Objective: To study the induction of human MCF-7 breast cancer cell apoptosis by inotodiol isolated from Inonotus obliquus, a traditional Chinese medicinal fungi. Methods: Inotodiol was isolated from the crude ethanol extract of I. obliquus and purified. Inotodiol at different concentration and human breast cancer MCF-7 cell line were co-incubated. Relative cell viabilities were determined by MTT method; apoptosis was analyzed by Annexin V/PI dual staining with flow cytometry; The protein expression levels of Caspase-3 and Poly ADP ribose polymerase (PARP) were detected by Western blotting. Results: Inotodiol reduced the viability of MCF-7 cells dose-and time-dependently. Moreover, inotodiol induced the apoptosis in MCF-7 cells. The data of Western blotting showed that inotodiol up-regulated the expression levels of cleaved Caspase-3 and PARP, in a time-dependent manner. Conclusion: Inotodiol could induce apoptosis in human breast cancer cells, which provides novel information for clinical application of inotodiol and its preparations for the breast cancer patients.

Triterpenoids of Inonotus obliquus / 中草药

Objective: To study the triterpenoids from the fruiting body of Inonotus obliquus. Methods: The compounds were isolated and purified by silica gel, MCI gel, Sephadex LH-20 column chromatographies, and their structures were mainly elucidated on the basis of physicochemical characteristics and spectral analysis. Results: Fifteen compounds were isolated from 95% EtOH extract of I. obliquus, and were identified as 3-oxo-lanosta-8, 24-diene-21-al (1), lanosterol (2), 3β-hydroxy-lanosta-8, 24-diene-21-al (3), betulin (4), inotodiol (5), trametenolic acid (6), 3β, 21-dihydroxy-lanosta-8, 24-diene (7), oleanic acid (8), ursolic acid (9), betulinic acid (10), inonotusane A (11), inoterpene D (12), 3-O-acetyl-11α, 12α-epoxy-oleanan-28, 13β-olide (13), ergosterol (14), and ergosta-4, 6, 8, 22-tetraene-3-one (15), respectively. Conclusion: Compound 1 is a new triterpenoid named as inonotusane D, while compounds 9, 13, and 15 are isolated from I. obliquus for the first time.

Ethanol extract of Innotus obliquus (Chaga mushroom) induces G1 cell cycle arrest in HT-29 human colon cancer cells

BACKGROUND/OBJECTIVES: Inonotus obliquus (I. obliquus, Chaga mushroom) has long been used as a folk medicine to treat cancer. In the present study, we examined whether or not ethanol extract of I. obliquus (EEIO) inhibits cell cycle progression in HT-29 human colon cancer cells, in addition to its mechanism of action. MATERIALS/METHODS: To examine the effects of Inonotus obliquus on the cell cycle progression and the molecular mechanism in colon cancer cells, HT-29 human colon cancer cells were cultured in the presence of 2.5 - 10 microg/mL of EEIO, and analyzed the cell cycle arrest by flow cytometry and the cell cycle controlling protein expression by Western blotting. RESULTS: Treatment cells with 2.5 - 10 microg/mL of EEIO reduced viable HT-29 cell numbers and DNA synthesis, increased the percentage of cells in G1 phase, decreased protein expression of CDK2, CDK4, and cyclin D1, increased expression of p21, p27, and p53, and inhibited phosphorylation of Rb and E2F1 expression. Among I. obliquus fractions, fraction 2 (fractionated by dichloromethane from EEIO) showed the same effect as EEIO treatment on cell proliferation and cell cycle-related protein levels. CONCLUSIONS: These results demonstrate that fraction 2 is the major fraction that induces G1 arrest and inhibits cell proliferation, suggesting I. obliquus could be used as a natural anti-cancer ingredient in the food and/or pharmaceutical industry.