The Optimization Of Ganoderma Lucidum Fermentation Condition And Immune Regulating Effects To Immunosuppressive Mice

This experiment changed the concentration of the Ganoderma lucidun liquid in the domestication culture medium to inoculating Bacillus subtilis, and screen out the suitable concentration of Bacillus subtilis growing. Taking the extracellular polysaccharide content as indexes, the test selected the inoculation quantity, stirring speed, initial pH, fermentation temperature and fermentation time five experiment factors to optimize the fermentation condition. The structure change of Ganoderma lucidum power after fermentation was observed by using HIT A CHI S-3400 N scanning electron microscope.And the fermentated Ganoderma lucidum polysaccharide and common Ganoderma lucidum polysaccharide were scanned by Bruker Tensor 27 infrared spectrometer to observe the absorption peak.This experment selected 80 mices, and randomly divided into 8 groups, each group of 10 mices. Except the blank control group, the rest of groups were injected cyclophosphamide of 80mg/kg for 3 days continuously to build immunosuppressive mouse models. At the fourth day, two kinds of the high-,medium-, low-dose Ganoderma lucidum polysaccharide groups were given to mices with different contents of 200, 100, 50mg/(kg?d) corresponding polysaccharide solutions, the blank control group and experimental control group were given correspondin volume of normal saline, once a day, and given 0.1mL/10 g dose in 7 days continuously. And at 24 h after the last administration in the end of the 10 th day,killed the mices, detected the organ indices; using the ELISA method to determine the concentrations of IL-2, IL-4, IFN-γ, TNF-α, IgG, IgM and IgA in serum; and using ultraviolet spectrophotometer to detect the levels of liver glutathione peroxidase(GSH-Px), super-oxide dismutase(SOD), total antioxidant capacity(T-AOC), catalase(CAT) and maleic dialdehyd(MDA).Results: The experiment found that the Bacillus subtilis could stably grow in the 14%concentration of the Ganoderma lucidun, but could not exist stably when the the concentration of the Ganoderma lucidun liquid was 16% liquid in liquid domestication culture medium. The best conditions of produced Ganoderma lucidum exopolysaccharide were 4% inoculation quantity, 180r/min stirring speed, 6.0 initial pH, 30℃ fermentation temperature, and 16 fermentation time. By scanning electron microscopy(SEM), the results showed that the structure of fermentated Ganoderma lucidum power was collapsed, crushed into fibrous, and existed in hyphal fragments scattered; but the structure of control sample still existed quite a certain lot of bacteria meat hyphae organization, its structure was not completely destroyed, maintained a certain integrity. Through the scanning by infrared spectrometer, the results showed that the most of absorption peaks were in the similar wavenumber range of the two kinds of Ganoderma lucidum polysaccharides, but there had absorption peak in the wavenumber range of3000~2800 cm-1in the fermentated Ganoderma lucidum polysaccharide, and the common Ganodermalucidum polysaccharide almost could not see absorption peak that caused by C-H key stretching vibration in this range, also the location, quantity and shape of the infrared spectrogram had a certain degree of changes of Ganoderma lucidum polysaccharide after fermentation. Compared with the blank control group, the spleen and thymus indexs were very significantly reduced, the concentrations of IL-2,IL-4, IFN-γ, TNF-α, IgG, IgM and IgA in serum, and the levels of liver GSH-Px, SOD, T-AOC, CAT were very significantly decreased, but the level of liver MDA was very significantly increased in the experimental control group(P<0.01). And compared with the experimental control group, the thymus index was significantly increased in the low-dose fermentated Ganoderma lucidum polysaccharide group(P<0.05), but was very significantly increased in the medium-,high-dose fermentated Ganoderma lucidum polysaccharide groups(P<0.01). The spleen index, the concentrations of IL-2, IL-4, IFN-γ,TNF-α, IgG, IgM and IgA in serum, and the levels of liver GSH-Px, SOD, T-AOC, CAT were very significantly increased, but the level of liver MDA was very significantly decreased(P<0.01) in each dose fermentated Ganoderma lucidum polysaccharide group in immunosuppressed mice. As the same time, the effects of the immunoregulatory and antioxidant fuction of the fermentated Ganoderma lucidum polysaccharide had obvious enhanced than the common Ganoderma lucidum polysaccharide.Conclusion: Used the domesticated Bacillus subtilis to fermente Ganoderma lucidum, through screening of the five signal factor, optimized the best fermentated technology of Ganoderma lucidum exopolysaccharides yield. The experiment found that the bacteria meat hyphae organization of fermentated Ganoderma lucidum power was collapsed, most existed in hyphal fragments scattered, and could not keep complete form structure. And the test found that the functional groups of fermentated Ganoderma lucidum polysaccharide had a certain degree of changes. Compared with common Ganoderma lucidum polysaccharide, the effects of immunoregulatory and antioxidant function had significantly enhanced with the fermentated Ganoderma lucidum polysaccharide.