Study On The Pharmacologic Activity And Toxicological Evaluation Of Polysaccharide From Fermentation Extract Of Inonotus Obliquus

Inonotus obliquus (Fr.) pilat is a very rare and valuable medicinal fungus, it is called Chaga in Russia, the conventional medicinal part is the sclerotia. From the16-17th century, Inonotus obliquus has been used to prevent and control various intractable diseases by folk in the Eastern Europe and Russia, such as digestive system inflammation, heart disease, cancers and so on. In traditional application, most of the active components of Inonotus obliquus were extracted from the fruit body. But the wild resources were scarce and the large-scale artificial cultivation was still difficult to achieve, so the liquid fermentation cultivation of Inonotus obliquus had the advantages of short production cycle and low costs, thus, it was signaficant to develop and utilize Inonotus obliquus. In this study, the fermentation extract polysaccharide of Inonotus obliquus was abtained through the liquid fermentation cultivation by fermentors, and the effects including anti-inflammatory、analgesia、immunological hepatic injury and anti-aging of the fermentation extract polysaccharides of Inonotus obliquus were studied. It laid the foundation for deeply study of the medicinal fungi on the treatment of hepatitis and the improvement of physical function for aging population. Meanwhile, the toxicological evaluation was researched by the methods of acute toxicity test and genetic toxicity test, it supplied the safety basis for application in the area of edible and medicinal health products.The results were as follows:1. To study the anti-inflammatory effects including both acute and chronic inflammation of the fermentation extract polysaccharide of Inonotus obliquus, the mice modles were built of increasing abdominal capillary permeability, ear edema, paw edema and granuloma induced by acetic acid, xylene, carrageenan and cotton balls, respectively. The results showed that, the fermentation extract polysaccharide of Inonotus obliquus could significantly inhibit the elevation of capillary permeability (p<0.01orp<0.05); the highest inhibition rate in the modle of mice ear edema induced by xylene was40.79%; in the test of mice paw edema caused by carrageenan, it showed significant inhibitory effect to the mice paw edema in the3th,4th and5th hour (p<0.0lor p<0.05); all dose groups of the fermentation extract polysaccharide of Inonotus obliquus showed highly significant inhibitory effect to mice granuloma (p<0.01), and the inhibitory rate of the high dose group was52.10%.2. Writhing test and hot plate test were used to study analgesic effects of the fermentation extract polysaccharide of Inonotus obliquus. The results showed that, in the writhing test, all dose groups of the fermentation extract polysaccharide of Inonotus obliquus could significantly extend the pain incubation period and reduced the times of writhing, the inhibitory rate of the high dose group was48.38%, that showed a better effect than the middle and low dose group; in the hot plate test, all dose groups of the fermentation extract polysaccharide of Inonotus obliquus showed no significant inhibition to the pain in the feet of mice.3. To study the protection of the fermentation extract polysaccharide of Inonotus obliquus against the immunological liver injury induced by injecting Bacille-Calmette-Guerin (BCG) and lipopolysaccharide (LPS), the indexes of spleen, thymus and liver were measured. And the levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), interleukin-12(IL-12) and interleukin-18(IL-18) in serum were examined, as well as superoxide dismutase (SOD), malondialdehyde (MDA), tumor necrosis factor-a (TNF-α), interferon-y (IFN-γ) and nitric oxide (NO) in liver homogenates. Histopathologic changes of the liver tissues were analysed. The results showed that, each dose of the fermentation extract polysaccharide of Inonotus obliquus could effectively decrease the contents of ALT, AST, IL-12, IL-18in serum; reduce the level of NO, TNF-α, INF-γ in liver homogenates, inhibit the accumulation of MDA; and enhance the activity of SOD. In the morphology, the phenomenon of the liver cell degeneration and necrosis was significantly improved. In conclusion, fermentation extract polysaccharide had remarkable hepatoprotective effect on immunological liver injury. The mechanism might due to increasing the activity of antioxidant enzymes, reducing the level of lipid peroxides and cytokines, adjusting immune function and so on.4. To study the effects of fermentation extract polysaccharide of Inonotus obliquus on various aging indicators of mice treated with D-galactose. The indexes of immune organs, activities of CAT, SOD, GSH-PX, and contents of MDA in serum, liver and brain were chose as indicators. The results showed that, after6weeks, compared with model group, fermentation extract polysaccharide could resist immune organs atrophy, enhance the activity of CAT, SOD and GSH-Px, reduce the content of MDA in serum, liver and brain of aging mice, its optimum dosage was500mg/kg. In conclusion, fermentation extract polysaccharide had remarkable anti-aging effect.5. Acute toxicity test, the mice sperm deformity test and the micronucleus in bone marrow cells test were used to evaluate the acute toxicity and genetic toxicity of fermentation extract polysaccharide of Inonotus obliquus. The results showed that, in the acute toxicity test, the mice did not show any toxic reaction under the administered dose of24g/kg, indicated that the maximum tolerated dose of fermentation extract polysaccharide of Inonotus obliquus was more than24g/kg, exceed of the100times of possible intake of People, has no acute toxicity; in the mice sperm deformity test, compared with the negative group, the rate of deformity sperm of mice in the group of cyclophosphamide had significant differences, but the rate of deformity sperm of mice in the group of fermentation extract polysaccharide had no significant differences with the negative group, the test results were negative; in the micronucleus in bone marrow cells test, the rate of micronucleus in polysaccharide group had significant differences with the cyclophosphamide group, but had no differences with the negative group, the test results were negative. In conclusion, fermentation extract polysaccharide of Inonotus obliquus neither have acute toxicity, mutagenicity nor genetic toxicity.In conclusion, the fermentation extract polysaccharide of Inonotus obliquus showed well effects on anti-inflammatory, analgesia, immunological hepatic injury and anti-aging, had a good development prospect in hepatitis treatment and auxiliary treatment. The toxicological evaluation experiments also showed that fermentation extract polysaccharide of Inonotus obliquus neither have acute toxicity, mutagenicity nor genetic toxicity, meant that it has safety basis and guarantee in the development and utilization of health care field.