Study On The Effect And Mechanism Of Chinese Herbal Compound With Homology Of Medicine And Food On Prevention And Treatment Of Alcoholic Liver Injury

Alcoholic liver injury,also known as alcoholic liver disease(ALD),is a major health problem in the country and even the world.It is mainly caused by excessive alcohol intake,which leads to liver oxidative stress injury and immune injury.At the same time,alcohol and its metabolites destroy intestinal health and cause a double blow to the liver through the gut-liver axis.The process is considered to play a key role in the occurrence of ALD.At present,there is no specific drug with high safety that can prevent and treat ALD.However,Chinese medicine,especially Chinese medicine with homology of medicine and food,shows a good effect of antagonizing ALD.If scientific compatibility is combined,ALD can be prevented and treated in daily diet.Objective:Under the guidance of Chinese medicine theory,the compatibility of medicine and food homologous Chinese medicine compound(referred to as prescription).To explore the efficacy and mechanism of its prevention and treatment of ALD,and to provide a theoretical basis for the development of Chinese medicine food and health services and the prevention and treatment of ALD.Method:1.To develop a Chinese medicine compound of medicine and food homology for the prevention and treatment of ALD.The data of ancient prescriptions for the prevention and treatment of ALD in Chinese Medical Classics were mined by using the traditional Chinese medicine inheritance calculation platform,and the prescription medication rules of ancient Chinese medicine for the prevention and treatment of ALD were analyzed.On this basis,the compound prescription was formulated by using the theory of traditional Chinese medicine.2.Study on the effect and mechanism of Chinese medicine compound on the prevention and treatment of ALD in rats.The ALD rat model was established by gradient alcohol feeding method,and the prescription was given at the same time.The protective effect of the prescription on liver and intestine of ALD rats was evaluated from the perspective of general signs,serum biochemistry and histopathology.According to the intestinal mucosal barrier function and lipopolysaccharide(LPS)/ Toll-like receptor(TLR)4 pathway,the correlation between the mechanism of the prescription in preventing and treating ALD and the gut-liver axis was explored.3.The effect of Chinese medicine compound on intestinal microorganisms in ALD rats.Bioinformatics statistical analysis was performed on the cecal contents of ALD rats.According to the analysis of flora biodiversity,flora structure,biochemical index correlation and function prediction,the mechanism of compound prevention and treatment of ALD was further explored from the perspective of intestinal biological barrier.Result:1.To develop a Chinese medicine compound of medicine and food homology for the prevention and treatment of ALD.(1)A total of 240 prescriptions were screened,including 189 flavors of traditional Chinese medicine,mainly for tonifying deficiency,regulating qi and promoting diuresis and eliminating dampness.Most of the medications belong to the spleen,stomach and lung meridians.The medicinal properties are always warm,cold and flat,and the flavors are always acrid,bitter and warm.30 pairs of high-frequency drug combinations and 28 association rules were obtained by association analysis.Cluster analysis showed that class 1 dahuangzhizi-gancao-huangqin-yinchen,class 2 baizhu-fuling-gegen-chenpi-renshen and class 3 gegen-gehua-chenpi-gancao-sharen.(2)Class 2 and 3 are highly consistent with the composition of Dongyuan Gehua Jiecheng Decoction,and they are intended to be combined to form the prescription for the prevention and treatment of ALD.The prescription was developed: gegen 15 g,fuling 15 g,sharen 10 g,chenpi 10 g,dangshen 10 g,ganjiang 6g.2.Study on the effect and mechanism of Chinese medicine compound on the prevention and treatment of ALD in rats.(1)General signs: The prescription can improve the symptoms of ALD rats such as withered hair,mental malaise,slow response,loose stools,yellow urine,no desire to drink water,frequent scratching and drunkenness.(2)Serum biochemistry: Compared with the normal group,the white blood cell(WBC),lymphocyte number(Lymph #)and monocyte number(Mon#)in the model group were significantly increased(P<0.01),and serum alanine transaminase(ALT),aspartate transaminase(AST),total cholesterol(TC)and liver malondialdehyde(MDA)in the model group were increased(P<0.05).Compared with the model group,the WBC and Lymph # in the highdose prescription group decreased(P<0.05),the serum TC in the low-dose prescription group,the serum AST in the middle-dose prescription group,and the serum ALT and AST in the high-dose prescription group decreased significantly(P<0.01).(3)Pathological morphology: In the model group,inflammatory cell infiltration,hepatic cord disorder,local edema,nuclear disappearance,a large number of red lipid droplets,small intestinal villi on the inner wall of ileum,local atrophy and shedding were observed in the liver of rats.The inflammatory cell infiltration and red lipid droplets in the liver of the rats in the high-dose prescription group were reduced,and the intestinal villi were intact and densely arranged,which was close to the tissue morphology of the normal group.(4)Ethanol metabolic enzymes: Compared with the normal group,the concentration of serum acetaldehyde dehydrogenase(ALDH)in the model group decreased(P<0.05).Compared with the model group,the concentration of alcohol dehydrogenase(ADH)in the high-dose prescription group increased(P<0.05).(5)Intestinal permeability: Compared with the normal group,the serum LPS and D-lactic acid(D-LA)concentration in the model group was significantly increased(P<0.001,P<0.01),and the expression of occludin was decreased.Compared with the model group,the serum LPS and D-LA concentrations in the middle and high dose prescription groups decreased(all P<0.05), and the expression of occludin protein in ileum increased.(6)Serum inflammatory factors: Compared with the normal group,serum tumor necrosis factor-α(TNF-α)and interleukin(IL)-6 increased and IL-10 decreased in the model group(all P<0.05).Compared with the model group,the serum levels of TNF-α and IL-6 in the middle and high dose prescription groups were significantly decreased(P<0.01),and IL-10 was increased(P<0.05).(7)TLR4 pathway: Compared with the normal group,the protein expression of TLR4,myeloid differentiation factor 88(My D88)and nuclear factor kappa-B(NF-κB)p65 in the liver of the model group increased(all P<0.05).Compared with the model group,the expression of NF-κB p65 protein in the liver of rats in the low dose prescription group was significantly decreased(P<0.01),and the expression of TLR4,My D88 and NF-κB p65 protein in the liver of rats in the middle and high dose prescription groups was decreased(all P<0.05,all P<0.01).3.The effect of Chinese medicine compound on intestinal microorganisms in ALD rats.(1)Biodiversity: Compared with the normal group,the sobs,chao and ace indexes of the model group were significantly decreased(all P<0.001),the shannon index was decreased and the simpson index was increased(all P<0.05).Compared with the model group,the sobs and shannon indexes of the low-dose prescription group increased,the sobs,chao and ace indexes of the middle-dose prescription group increased(all P<0.05),and the sobs,chao,ace and shannon indexes of the high-dose prescription group increased(all P<0.05).(2)Phylum level flora structure: Compared with the normal group,the abundance of Proteobacteria in the model group decreased significantly(P<0.01).Compared with the model group,the abundance of Proteobacteria in the low and high dose prescription groups increased(all P<0.05),and the abundance of Proteobacteria and Deferribacterota in the middle dose prescription group increased significantly(all P<0.01).(3)Genus level flora structure: Compared with the normal group,the abundance of Psychrobacter in the model group was significantly decreased(P<0.01),and the abundance of Lachnospiraceae＿NK4A136＿group,Jeotgalicoccus and Blautia was decreased(all P<0.05).Compared with the model group,the abundance of Prevotellaceae＿NK3B31＿group,Colidextribacter,NK4A214＿group and Ruminococcus increased significantly in the low-dose prescription group(all P<0.05).The abundance of Desulfovibrio increased significantly in the middle-dose prescription group(P<0.01),and the abundance of UBA1819 and Anaerobacter increased significantly(all P<0.001).The abundance of norank＿f＿Muribaculaceae and unclassified＿f＿Ruminococcaceae in the high-dose prescription group increased significantly(all P<0.01),and the abundance of Desulfovibrio increased(P<0.05).(4)Specific flora: The normal group was unclassified＿f＿＿Lachnospiraceae and Blautia in the order of Lachnospiraceae.The model group was Turicibacter.The low dose prescription group was Ruminococcaceae and UCG-003,the middle dose prescription group was norank＿f＿＿Erysipelotrichaceae,and the high dose prescription group was norank＿f＿＿Muribaculaceae in Muribaculaceae.(5)Correlation of biochemical indicators: norank＿f＿＿Muribaculaceae was negatively correlated with AST,LPS and TNF-α;Desulfovibrio was negatively correlated with AST,ALT,TNF-α and IL-1β;norank＿f＿＿Oscillospiraceae was negatively correlated with ALT;Intestinimonas was negatively correlated with TNF-α;Prevotaceae＿UCG-003 was negatively correlated with AST.Norank＿f＿Eubacterium＿coprostanoligenes＿group was negatively correlated with AST,and Staphylococcus was positively correlated with D-LA(all P < 0.05).Conclusion:1.Ancient Chinese medicine for the prevention and treatment of ALD is always based on tonifying qi,regulating qi,relieving alcohol and dehumidification,and is compatible with common tonifying qi drugs-water disinhibititing drugs,tonifying qi drugs-regulating qi drugs,dampness drugs-regulating qi drugs and drugs with similar efficacy.Combined with the class 2,3 and Dongyuan Gehua Jiecheng Decoction,the prescription was developed: gegen15 g,fuling 15 g,sharen 10 g,chenpi 10 g,dangshen 10 g,ganjiang 6g.2.The prescription can protect the liver and intestinal health of ALD rats,improve the abnormal signs and symptoms of ALD rats,reduce serum liver function and blood lipid levels,and protect the tissue structure of liver and ileum.The mechanism may be :(1)Increase the concentration of ADH,accelerate ethanol metabolism.(2)Enhance intestinal mucosal barrier function,reduce endotoxin into the liver to activate TLR4 / My D88 / NF-κB pathway,thereby reducing the second hit to the liver.3.The prescription can increase the intestinal microbial diversity of ALD rats.The mechanism of improving intestinal mucosal barrier function and regulating liver immune injury is related to Proteobacteria and Deferribacterota at the phylum level,but the specific mechanism needs further study.At the genus level,it is related to increasing the relative abundance of norank＿f＿Muribaculac eae and Desulfovibrio.