Establishment Of Bidirectional Fermentation System Of Liquorice Residue And Pharmacodynamics Stud

With the rapid development of Chinese medicine pharmaceutical industry,therefore the volume of herb dregs produced by industrial production is huge.Meanwhile,the disorderly emissions of herb dregs not only cause environment damage but also huge economic loss.The widely application of Glycyrrhizae radix in clinical medicine leads to considerable quantity of Glycyrrhizae radix dregs,while Glycyrrhizae radix dregs still contains many active ingredients,such as polysaccharides,flavonoids,etc.Thus,the problem that how to deal with Glycyrrhizae radix dregs benefitly and harmlessly needs to be solved urgently.Bidrectional fermentation is called the medicianal fungal new type(bidirectional)solidstate fermentation engineering,which refers to that medicinal fungus and traditional Chinese medicine(TCM)fermentation in order to gain both attenuation of toxicity and synergistic effects of efficacy.It is the sublimation of TCM fermentation processing methods.Many bidrectional fermentation products exhibt satisfied effect on clinical treatment,such as Trametes robiniophila Murr-Radix Astragali fermentation product and Ganoderma lucidumTripterygium wilfordii fermentation product,these results proved that bidrectional fermentation prosess enormous application value.However,bi-directional fermentation is used only for fermentation of TCM materials and medicinal fungus,except for dregs of TCM.Thus,the application of bi-directional fermentation for the processing of dregs could generate enormous economic benefits and broad application prospects.Studies have shown that medicinal fungi,Trametes robiniophila Murr,Penicillium cicada and Ganoderma sinense are all have strong adaptability on TCM.Moreover,these three contain a variety of active ingredients,such as fungal polysaccharides,flavonoids,have a good antioxidant,immunomodulatory and antitumor efficacy.This thesis combine Trametes robiniophila Murr,Penicillium cicada and Ganoderma sinense respectively with Glycyrrhizae radix dregs,set up three kinds of bi-directional fermentation systems,select the optimal fermentation system by determining polysaccharides and flavonoids in the process of fermentation,then explore the composition change in the fermentation process,so that we can provide ideas and direction for other fermentation systems.Compare immune adjustment ability of fermented with un-fermented product in vitro using pharmacodynamics exploration,determine the optimal fermentation process by using orthogonal design,and then conduct a targeted in vivo pharmacodynamics study,so as to provide new efficient disposal ways of Glycyrrhizae radix dregs.The methods and results of this paper are as follows:1.Screening optional Glycyrrhizae radix dregs feimentation strainsCombine Trametes robiniophila Murr,Penicillium cicada and Ganoderma sinense respectively with Glycyrrhizae radix dregs,set up three kinds of bi-directional fermentation systems,determine substrate consumption rate to evaluate the adaptability of these three fermentation system,detect the change of polysaccharides and flavonoids content in three kinds of fermentation systems by using colorimetric method,determine the optimal fermentation strains and use antioxidant experiment for validation.The results are as follows:according to the substrate consumption rates,three kinds of fermentation systems are all feasible.During the late fermentation period,polysaccharides and flavonoids content of Penicillium cicada/Glycyrrhizae radix dregs are significantly higher than that of the raw Glycyrrhizae radix dregs.Free radical clearance rates and ferrous ion chelating ratess have confirmed that Penicillium cicada/Glycyrrhizae radix dregs has the best oxidation resistance among these three.In conclusion,Penicillium cicada is the optimal frmentation strains of Glycyrrhizae radix dregs.2.Determination of component content changes of Penicillium cicada/Glycyrrhizae radix dregs system during the fermentation processIn order to reveal the components change rule of fermentation process,this paper determined the change of flavonoids ingredients(liquiritin,isoliquiritin,liquiritigenin,isoliquiritigenin)and triterpenoid saponins class composition glycyrrhizic acid of the optimal fermentation system(Penicillium cicada/Glycyrrhizae radix dregs)by using high performance liquid chromatography(HPLC).The results are as follows:flavonoid glycoside compounds significantly reduced while glycosides metaclass composition increased,which means that there’s existing material transformation during the fermentation process.At the same time,glycyrrhizic acid which has efficacy liver protection,anti-inflammatory,detoxification and antitumor increased.In conclusion,after fermentation,the type and concentration of active ingredients changed,prompting that bidirectional fermentation of Glycyrrhizae radix dregs has positive significance,and also provide new way for it’s dispose.3.Exploration of in vitro immune adjustment ability of Penicillium cicada/Glycyrrhizae radix dregs fermentation productTo explore the quality of Penicillium cicadalGlycyrrhizae radix dregs fermentation product and the application prospect,this thesis take polysaccharide as the research object,determine polysaccharides on macrophage proliferation and phagocytosis ability.Results are as follows:polysaccharides showed a certain quality of promoting proliferation and macrophage function,and the effects of the polysaccharide in the fermented 13 days is the strongest.It suggests that the fermentation product has a strong ability of immunity regulation and determine the fermentation end-point is 13 d.4.Determination of optimal fermentation process of Penicillium cicada/Glycyrrhizae radix dregs bi-directional fermentation systemPolysaccharide as index to optimize the optimal fermentation using the method of orthogonal design.The optimal fermentation conditions are as follows:relative humidity is 90%,fermentation temperature 28℃,inoculation amount of 3 mL/5 g,water content of 4 mL/5 g.5.Exploration of in vivo immune adjustment ability of Penicillium cicada/Glycyrrhizae radix dregs fermentation productCarry out the in vivo efficacy experiment to evaluate immune modulate ability of polysaccharide,through determination of polysaccharides on the immunodeficiency mice immune organ index and level of serum immune factors(IL-2,TNF-α,IL-12)to evaluation of immunity ability of Penicillium cicada/Glycyrrhizae radix dregs bi-directional fermentation system.Results show that polysaccharide can significantly improve immune organs index and the concentration of IL-2 and TNF-α factors,and there are significant differences between the experimental group and model group(P<0.01),which suggesting the polysaccharide can improve the immune function of mice,and this result laying a foundation for the industrial development of Glycyrrhizae radix dregs.