Study On Chemical Constituents And Pharmacological Activity Of Inonotus Obliquus

The mushroom Inonotus obliquus (Fr.) Pilát (IO),which name is Chaga in Russian, is a black parasitic fungus that grows on living trunks of the mature birch, elm, alder or beech trees. It is mainly found at latitudes of 45?N–50?N, such as Western Siberia, Poland, Finland,Changbai Mountain and Heilong River in China. Traditionally, it has been used for the treatment of gastrointestinal cancer, cardiovascular disease and diabetes since the 16th century in Russia, Poland and most of the Baltic countries. However, its chemical components and pharmacological actions have not been well documented, in spite of its increasing usage.A comprehensive analytical study on the chemical constituents of IO was performed. The stepwise extractions of IO were performed using petroleum ether, acetone, methanol and water orderly. The main chemical constituents in the extract with petroleum ethe(rIOPE) and extract with acetone(IOA) from IO were analyzed by GC-MS. 25 chemical constituents (relative content: 95.6%) of IOPE were identified by using GC-MS. The primary component were 6 steroids (relative content: 91.6%). 58 chemical constituents (relative content: 86.6%) of IOA were identified by using GC-MS. The primary components were 14 steroids (relative content: 77.1%).Several factors that affecting the extraction process of polysaccharide were studied through single factor and orthogonal experiments. The optimal technological conditions of polysaccharide extraction from IO were decided: the extraction time being 240 minutes; the temperature being 90 oC; the ratio of solid material to water being 1:14; the times of extraction being 7. 4 purified polysaccharides and 4 purified acid polysaccharides were prepared with process as the following: extracting, ethanol precipitating, Sevag method deproteinizing, DEAE-cellulose chromatographic fractionating, sephadex chromatographic purifying.The structure of Polysaccharide IOPS-2 was analyzed. Molecular weight of IOPS-2 estimated to be 2.76×104 detected by gel chromatography. IOPS-2 has been investigated by GC analysis, periodate oxidation, Smith degradation, methylation and partial acid hydrolysis analysis. The results showed that it is a glucomannan, composed of terminal Glcp, (1→3)-linked Glcp, (1→3)-linked Manp, (1→4,6)-linked Manp, (1→4,6)-linked Glcp in the molar ratios of 1.0: 2.5: 8.3: 3.2: 0.5.The pharmacological activities of extracts from IO (IOPE, IOA, IOM and IOW ) were studied. In vivo, the test showed that the extracts of IO have inhibition effects on tumor, the inhibition ratio increased with the polar of solvent. In vitro test, the result show that IOPE and IOA have cell toxicity on tumor cell, at the same time the IOW have no cell toxicity. In vitro test, the results also show that IO extracts containing serums have cytotoxic activity on K562 cell. The higher polarity the extract solvents have, the higher activity the serums have.To study the immunoregulatory effects of IO extracts, the results show that the extracts can increase the spleen index and thymus index of tumor-bearing mice, induce the proliferation of spleen lymphocytes, and increase the activities of NK and LAK cell.