Effects Of Pueraria Biofermentation Products On Alcoholic Fatty Liver Under Different Administration Methods

In recent years,the incidence rate of alcoholic fatty liver disease(AFLD)showed an upward trend around the worldwide,and there is no specific therapeutic drug at present.Pueraria lobata(P.lobata)is the dried root of pueraria legume.Its main bioactive substances include puerarin,daidzein,daidzein and other isoflavones,which have a wide range of pharmacological activities.P.lobata has been shown to relieve alcohol-related diseases,reduce oxidative stress,inflammatory response and apoptosis caused by AFLD,and has a good liver protection effect.Probiotics,as active biologics,promote host absorption of nutrients,enhance immunity,and protect liver cells from damage by inhibiting oxidative stress and inflammatory responses.At the same time,the probiotic fermentation processes could lead to decomposition or transformation of complex substrates,thereby improving product properties or altering the amount of certain bioactive compounds.In the early stage of Chinese traditional medicine,fermentative herbs such as Liushen Qu,Pinellia Qu and Pien Tze Huang have been proved to have good pharmacological activity.Recently,the production and enrichment of bioactive metabolites with medicinal value to improve herbal medicines by fermentation transformation has attracted wide attention worldwide.In addition,in terms of drug delivery,in addition to oral,injection,transdermal absorption and other traditional methods,atomized inhalation drug delivery has been widely used due to its advantages of high bioavailability,convenient drug delivery and good portability,but it is still in a blank state in the treatment of AFLD.In this thesis,different Pueraria raw materials and probiotics were screened to prepare Pueraria fermentation,and the alleviating effects of oral and atomized inhalation administration on AFLD were investigated.Specific research contents and results were as follows:(1)Boiling extraction,enzymatic hydrolysis extraction and compound extraction(enzymatic hydrolysis extraction followed by boiling extraction)were used to prepare the extracts of P.lobata and Pueraria thomsonii.The contents of puerarin,total flavonoids,total polyphenols and total sugars were determined.Extracellular metabolites of 15 strains of probiotics including Phellinus igniarius and Lactobacillus plantarum were prepared,and the above strains were used for fermentation and transformation of P.lobata,and then the scavenging ability of DPPH,superoxide anion and hydroxyl radical of Pueraria extract,extracellular metabolites of probiotics and P.lobata fermentation was analyzed.The results showed that: The antioxidant capacity of puerarin,total flavonoids and total polyphenols were the highest,which were increased by 31.86%,16.58% and 32.76% compared with traditional boiling extraction technology.The extracellular metabolites of Poria cocus,Ganoderma lucidum,Cordyceps militaris,Ganoderma sinense,Phellinus igniarius,Boletus edulis,Lactobacillus plantarum,Lactobacillus paracasei and Lactobacillus rhamnosus showed good free radical scavenging ability,and the free radical scavenging ability of the above strains was further enhanced after fermentation of P.lobata,reaching more than 50%.(2)Hep G2 cell injury model was induced by 200 mmol/L ethanol,and the repair effects of P.lobata fermentation on Hep G2 cell injury were analyzed.The results showed that: The combined fermentation product FPS138 of P.lobata by Phellinus igniarius(F-C13-1)and Lactobacillus plantarum(F-B8-1)at 0.5 mg/m L could significantly improve the survival rate of Hep G2 cell injury model and reduce ALT,AST and intracellular ROS levels,showing a good damage repair effect.Subsequently,highperformance liquid chromatography and colorimetry were used to determine the changes in the products before and after the fermentation of Pueraria lobata by Phellinus igniarius and Lactobacillus plantarum.The results showed that the content of puerarin,total flavonoids and total polyphenols in the combined fermentation product FPS138 increased by 34.73%,40.31% and 43.12% compared to before fermentation,respectively.(3)An AFLD mouse model was constructed using ethanol gavage.The alleviative effects of gavage(0.1 m L/10 g body weight)and inhalation administration(suction volume of 55 m L;suction frequency of 240 s/time)on the model mice were analyzed.The results showed that the high-dose FPS138 group significantly reduced the levels of serum ALT,AST,TG,TC,inflammatory factors,as well as the content of MDA,TC,TG in liver tissue,and increased the enzyme activity of SOD and GSH-PX.The results of histopathology showed that FPS138 could significantly repair the liver injury of model mice.RT-PCR transcriptional level analysis also showed that FPS138 had a significant regulatory effect on the expression of genes related to ethanol metabolism and lipid metabolism in model mice.In summary,this thesis analyzed the alleviative effects of FPS138 on AFLD mice through oral and inhalation administration methods,and demonstrated that the P.lobata fermented products treated with the combination of Phellinus igniarius(F-C13-1)and Lactobacillus plantarum(F-B8-1)have good effects in antioxidant,regulating lipid metabolism disorders,regulating ethanol metabolism,antagonizing liver injury and inhibiting inflammation,providing a new approach for the treatment of AFLD.