Synergistic Protective Effect Of Eurotium Cristatum Powder And Quercetin On Acute Alcohol-induced Liver Injury In Mice

Alcoholic liver injury caused by acute alcohol consumption is an important factor causing alcoholic liver disease(ALD),but the current clinical treatment of ALD drugs due to side effects or poor treatment efficacy and other shortcomings,so it is particularly important to find safe and effective drugs.Eurotium Cristium(EC)is a unique probiotics in Fu brick tea and Eurotium Cristium Fermented Loose Dark Tea,commonly known as "Golden Flower".The Eurotium cristatum powder(ECP)has not been reported in acute alcoholic liver injury(ALI).Quercetin(QC)contained in tea has a good therapeutic effect on ALI,but the synergistic effect of ECP and quercetin in ALI has not been reported.Therefore,in this study,we used ECP and quercetin and their combined pretreatment on oxidative stress,inflammatory reaction and detoxification in the liver of mice fed with acute alcohol.The survival status and pathological sections of liver tissue of mice were observed.The levels of liver function indexes ALT,AST and liver oxidative stress indexes ROS,MDA,GSH-Px,GSH and SOD were detected by biochemical kit.The levels of serum inflammatory factors TNF-α,IL-1 β,IL-6 and IL-10 were detected by ELISA method.The protein levels of Nrf-2,HO-1,CYP2E1,NF-κB,PXR,Ah R,CYP1A2 and CYP3A4 were detected by Western Blot.To explore the effect and mechanism of ECP alone or in combination with quercetin on ALI.In addition,metabonomic analysis was carried out on Eurotium Cristium Fermented Loose Dark Tea and its surface powder in order to provide a reference basis for the effect of Eurotium Cristium powder on ALI.The results are as follows:1.There are 212 kinds of differential metabolites between Eurotium Cristium Fermented Loose Dark Tea and Eurotium Cristium powder.The metabolomics analysis showed that there were more kinds of phenols and their derivatives,phenolic acids,glycosides,flavonoids,lignins and terpenes in Eurotium Cristium Fermented Loose Dark Tea.There are more kinds of amino acids and their derivatives,organic acids,alkaloids and sugars in Eurotium Cristium powder.They have the same number of esters and coumarins and their derivatives.2.After pretreatment with Eurotium Cristium powder or quercetin or the combination of both,the quality of life,liver function index and liver tissue pathological changes of acute drinking mice were significantly improved.In the acute drinking model mice,the body weight decreased sharply,the liver index,ALT and AST increased significantly,and the liver pathological changes were aggravated.Each drug prophylaxis treatment group improved these conditions in varying degrees,but there was no dose dependence,and the synergistic group showed better therapeutic effect.3.After pretreatment with Eurotium Cristium powder or quercetin or the combination of both,the serum inflammatory factors and liver oxidative stress indexes of acute drinking mice were significantly improved.In acute drinking model mice,the contents of ROS and MDA in liver and the levels of TNF-α,IL-1 β and IL-6 in serum increased significantly,while the levels of GSH-Px,GSH,SOD in liver and IL-10 in serum decreased significantly.Each drug prevention treatment group improved these conditions in varying degrees,and the antioxidant capacity of Eurotium Cristium powder was better than that of quercetin in a dose-dependent manner,which basically reached the effect of silymarin positive control group.It is worth noting that the synergistic group showed dose dependence in antioxidant and anti-inflammatory effects,and significant synergistic effect in oxidative stress indexes,but no obvious synergistic effect in inflammatory factors.this may be due to the fact that the effect of using Eurotium Cristium powder or quercetin alone is close to the normal level,and the synergistic effect is not obvious because the therapeutic effect is too good.4.After pretreatment with Eurotium Cristium powder or quercetin or the combination of both,the expression of CYP2E1 and NF-κ B protein decreased,while the expression of Nrf-2,HO-1,PXR,Ah R,CYP1A2 and CYP3A4 protein increased in acute drinking mice.In Nrf-2/HO-1/CYP2E1 and NF-κ B/Ah R/CYP1A2 signal pathway,Eurotium Cristium powder showed better ability to inhibit CYP2E1 enzyme activity and promote the increase of Nrf-2,HO-1,Ah R and CYP1A2 protein levels in a dose-dependent manner,while quercetin showed a better ability to promote the increase of PXR and CYP3A4 protein levels in NF-κB/PXR/CYP3A4 signal pathway in a dose-dependent manner.There was no significant difference in the ability of inhibiting NF-κB transcription factor activity between the two groups,and there was no significant dose dependence.The synergistic group had obvious synergistic effect on Nrf-2/HO-1 and Ah R/CYP1A2 signal pathway in a dose-dependent manner.The above results suggest that Eurotium Cristium powder and quercetin can improve the oxidative stress,inflammation and toxicity of liver induced by ALI,reduce oxidative stress by affecting Nrf-2/HO-1/CYP2E1 pathway,reduce inflammation by affecting NF-κ B/ Ah R and NF-κB/PXR pathway,and reduce hepatotoxicity by affecting PXR/CYP3A4 and Ah R/CYP1A2 pathway.The synergistic use of Eurotium Cristium powder and quercetin enhanced the influence of Nrf-2/HO-1 and Ah R/CYP1A2 signal pathway and had better pharmacological health care function.This study has opened the first page of the blank research field of Eurotium Cristium powder in acute drinking liver injury,and studied the synergistic mechanism of Eurotium Cristium powder and quercetin contained in Fu brick tea and Eurotium Cristium Fermented Loose Dark Tea on acute drinking liver injury.It not only provides the experimental basis for innovative drugs for the treatment of alcohol-induced liver injury,but also provides reference significance for Fu brick tea and Eurotium Cristium Fermented Loose Dark Tea with high quercetin content to prevent acute alcohol-induced liver injury.