Hepatoprotect I Ve Effect Of Extracts From Large Yellow Tea Based On Effective Microbial Fermentation On Alcohol Ic Liver Injury In Mice

Large Yellow Tea was a lightly fermented tea with a strong burnt flavor and produced in Huoshan county in Anhui province and Yingshan county in Hubei province.In addition to its unique tea soup flavor and sweetness,inexpensive Large Yellow Tea was rich in tea polyphenols,tea polysaccharides,proteins,which have certain nutritional value and pharmacological activity In this study,Large Yellow Tea was combined with effective microorganism(EMs)and fermented by EMs,and the extractive of effective microorganism fermented of Large Yellow Tea(FLYT)was obtained.The antioxidant capacity of FLYT was evaluated by antioxidant experiments in vitro,and an acute liver injury model was developed to evaluate the liver protection of FLYT in vivo.The main research contents were as follows1.Preparation and extraction process of FLYT:FLYT was obtained after crushing,EMs solid state fermentation,ethanol pretreatment,hot water extraction,vacuum concentration,centrifugation,ethanol precipitation,dissolution,vacuum concentration and freeze-drying Response surface methodology was designed to obtain the optimal technological parameters for FLYT extraction:the extraction time was 2.2 h,the temperature was 78℃,and the liquid-solid ratio was 21:1.The actual yield under the optimal condition was 6.69±0.59%.The degree of influence of three factors on FLYT yield was:Liquid-solid ratio>extraction time>extraction temperature2.The main components changes after the fermentation by EMs and the antioxidant ability of FLYT:Phenol-sulfuric acid method,Bradford method and Folin method were definited the main components of FLYT and the extractive of Large Yellow Tea(LYT),which were carbohydrates,proteins and polyphenols.The test results were as follows:the sugar content was 28.76%,protein content was 38.79%,and tea polyphenols content was 13.78%in LYT;the carbohydrates content was 40.79%,the proteins content was 28.93%,and the tea polyphenols content was 12.14%in FLYT.The results showed that the carbohydrates in FLYT were increased by EMs fermentation,while the proteins and polyphenols in FLYT were decreased High Performance Liquid Chromatography showed that the proportion of polysaccharides with smaller molecular weights was increased in FLYT.Antioxidant experiments showed that FLYT had excellent scavenging rate for DPPH,OH-,and ABTS+.At the dosage of 150μg/mL,FLYT can clear more than 80 percent for DPPH,OH-and ABTS+,and the scavenging rates was significantly higher than LYT in vitro3.Protective effect of FLYT on alcohol-induced acute liver injury:in this study,mice were given 12 mL/kg body weight 50%alcohol after continuous gavage of FLYTL,FLYTM and FLYTH for 30 days to establish an acute liver injury model.Serum levels of triglycerides(TG),total cholesterol(T-CHO),low density lipoprotein cholesterol(LDL-C)and high density lipoprotein cholesterol(HDL-C)were measured to evaluate the therapeutic effect of FLYT on alcohol-induced liver lipid metabolism disorders;The content of aspartate aminotransferase(AST),alanine aminotransferase(ALT)in serum and liver pathological sections were measured to evaluated the treatment effect of FLYT on alcohol-induced liver injury;In order to further determine the mitigation effect of FLYT on alcoholic liver injury,the content of alcohol metabolic enzymes,antioxidant enzyme,lipid peroxic product malonaldehyde(MDA),and the relative content of inflammatory factors were detected in this experiment.The experiment results show that FLYT could significantly relieve the lipid metabolism disorder caused by alcohol in the liver of mice.Under the treatment of FLYT,the alcohol metabolizing enzyme and the antioxidant enzyme activity in the liver of mice were increased,and the inflammatory state in the mice caused by alcohol was reduced in the liver of mice.In addition,histopathology has shown that FLYT can significantly alleviate alcohol-induced liver damage.At the same time,FLYT was more effective than LYT in protecting against alcohol-induced liver damage at the same dose4.FLYT’s protective potential mechanism against alcohol-induced acute liver injury in mice:the relative expression of nuclear factor erythroid-2 related factor 2(Nrf2),the catalase(CAT),Cu-Zn superoxide dismutase(Cu/Zn-SOD)and quinone oxidoreductase-1(NQO-1)was detected.It turns out that FLYT can activate Nrf2 and up-regulate downstream the level of CAT,Cu/Zn-SOD and NQO-1.