| Backgroud:Fatty liver is a clinicopathic syndrome in which the liver is abnormally metabolized by lipids,resulting in a large accumulation of fat in liver tissue.The main body of the lesion is in the hepatic lobules,with hepatic steatosis and fat accumulation as the main changes.According to the relationship between its onset and drinking,it can be divided into two major categories:alcoholic fatty liver and nonalcoholic fatty liver.With the development of the disease,fatty liver can develop from the initial simple fatty liver to hepatitis,and if it continues to increase,it will be liver fibrosis.In severe cases,cirrhosis can occur and eventually become hepatocellular carcinoma.At present,the incidence of fatty liver is increasing with people's lifestyle and eating habits,which is especially important for the diagnosis and treatment of fatty liver.The research on the pathogenesis of fatty liver in modern medicine has not been fully elucidated.Clinical treatment often needs to address different causes and causes of diseases,treat primary diseases and prevent complications.Traditional Chinese medicine treatment of fatty liver is guided by the overall concept,based on syndrome differentiation and treatment as the basic principle.According to the patient's clinical symptoms and physical signs,the corresponding prescriptions are applied for treatment.As long as the syndrome is accurate,the prescription can be used properly.Traditional Chinese medicine is a natural medicine with few side effects.Each traditional Chinese medicine contains a variety of active ingredients,thus providing a sufficient Chinese medicine library for the treatment of fatty liver.In recent years,a large number of clinical and pharmacological studies have also shown that a large number of single-flavor Chinese medicine and traditional Chinese medicine compound prescriptions are effective for fatty liver and its complications.Huangqi Gegentang originated from the Qing Dynasty medical book"Zhengzhihuibu",the prescription of Huangqi one or two,Gegen five money,the main qi deficiency and injury,the ancient doctors believe that the party has hangover effect.The previous animal experiments in this group have proved that Astragalus,Radix Puerariae and its main active ingredients,astragaloside ?,puerarin and their compatibility can effectively alleviate acute alcoholic gastric mucosal injury in rats.Astragalus and Radix Puerariae have also been reported in other treatments for fatty liver.In this study,we constructed a model of acute alcoholic liver injury in SD rats and a model of non-alcoholic liver injury induced by knockout mice(Sesn1/2/3 knockout)induced by HFHFC diet.The prevention and treatment of formazan and puerarin and the pathogenesis of fatty liver.Method:1.Establishment of an acute alcoholic liver and gastric injury model Female SD rats of 180-200 g were randomly divided into four groups,normal saline group(control group),alcoholic gavage group 1 time,alcoholic gavage 3 times group,and alcohol gavage 10 times group.With absolute ethanol or normal saline(control group),according to 5 ml/kg body weight,once a day,3 times(12h/time),10 times(12h/time).Gastric mucosa changes were observed with the naked eye,and the gastric mucosal injury score was calculated by Guth method.Finally,HE staining was used to observe the damage of gastric mucosa and liver tissue.2.Protective effect of astragaloside and puerarin on rats with acute alcoholic liver injury180-200g female SD rats were randomly divided into normal group,model group,astragaloside group,puerarin group,astragaloside combined with puerarin group.Each group was intraperitoneally injected for 4 days according to different drugs.The normal group was given normal saline(10ml/kg),the model group was given 10%propylene glycol:ethanol cosolvent(10ml/kg),and the astragaloside group was given 2mg/kg astragaloside ?.(Prepared astragaloside ? solution 0.2mg/ml according to 10ml/kg injection),puerarin group 60mg/kg puerarin(formed puerarin solution 6mg/ml according to 10ml/kg injection),astragaloside combined with puerarin group Astragaloside 2mg/kg and puerarin 30mg/kg solution(prepared with astragaloside 0.2mg/ml puerarin 30mg/ml solution,injected according to 10ml/kg).The changes of gastric mucosa were observed by the naked eye.The gastric mucosal injury score was calculated by Guth method.The pathological changes of gastric mucosa and liver tissue were observed by HE staining.The liver function damage of rats in each group was evaluated by serum AST and ALT.The liver mitochondria of each group were extracted.Immunoblot and real-time PCR were used to detect the expression of alcohol-related gene CYP2E1 and mitochondrial homeostasis Mortalin,and to explore the possible mechanism of astragaloside and puerarin in the prevention and treatment of alcoholic liver and stomach injury.3.Regulation and mechanism of Sesns in a diet-induced fatty liver mouse modelHigh-fat,high-glycocholate diet-induced fatty liver model in mice:normal group(WT group)and liver-specific knockout Sesn1/2/3 gene group(TKO group),common chow diet and HFHFC diet was used to observe the liver function of mice in each group after 8 weeks.Liver pathological changes were observed by HE staining and Sirius staining.Lipid metabolism,liver fibrosis and oxidation were detected by real-time PCR.Expression of related genes such as stress and inflammatory factors,WB method was used to detect the expression of related proteins,ELISA was used to detect the expression levels of oxidase and antioxidant enzymes,and the formation of fatty liver induced by high-fat,high-glycocholate diet was explored by Sesns knockout.influences.In vitro,the Sex and Mapk9 plasmids were co-transfected into LX-2 cells,and the Sesn and Mapk9 genes were overexpressed.The interaction between protein and protein was observed by CO-IP method,and the possible mechanism of the effect of Sesns on fatty liver regulation was studied.Result:1.Rat model of acute liver and stomach injury,the experimental results show that after 10 times of alcoholic administration,the mortality of rats increased significantly.It is not suitable to use this model for later experimental research.Gastric mucosal injury and liver injury increased slightly after 3 times of intragastric administration,but there was no statistical difference between the two groups.Therefore,the model of acute liver and stomach injury in rats was intragastrically administered with 5 ml/kg body weight of absolute ethanol,and a later experiment was carried out.2.Astragaloside and puerarin have protective effects on acute alcoholic liver injury in rats.The experimental results show that compared with the model group,2 mg/kg astragaloside ? solution,60 mg/kg puerarin solution,and astragaloside 2 mg/kg There were significant differences in gastric mucosal injury score,liver HE staining,liver function ALT,AST and other indicators in the three groups of rats treated with puerarin 30 mg/kg,suggesting that all three groups can protect rats from acute alcohol.Liver function damage can reduce liver CYP2E1 mRNA level and up-regulate liver mitochondrial CYP2E1 and Mortalin protein expression,which may be one of the protective mechanisms.3.The fatty liver model was induced by high fat and high cholesterol diet(HFHFC diet).The results showed that compared with WT group,liver Sesn1/2/3 knockout mice(TKO group)can promote the formation and development of fatty liver induced by HFHFC diet..The results of HE staining in liver sections of TKO group showed that hepatic steatosis aggravated and inflammatory cells accumulate;Sirius staining suggested that collagen fiber formation increased;serum ALT,liver tissue protein homogenate TG,TC results indicated hepatocyte injury aggravated,blood lipid levels rise.PCR results showed that oxidative stress,proinflammatory factors and liver fibrosis gene expression were upregulated.It is suggested that the Sesns gene knockout may be involved in the formation and development of fatty liver by regulating the "second strike" factors of non-alcoholic fatty liver formation,such as oxidative stress,inflammatory factors and extracellular matrix.LX-2 cells were co-transfected.The results of CO-IP assay showed that Sesn gene and Mapk9 gene interacted with protein and protein,suggesting that Sesns may participate in the formation and development of fatty liver at the molecular level by regulating JNK pathway.Conclusion:1.SD rats were given a single oral gavage with absolute ethanol 5ml/kg,which can cause acute alcoholic liver and stomach in rats.damage.This model can be used as a model tool for studying the effects of drugs on acute alcoholic liver and stomach injury.2.2mg/kg astragaloside ?,60mg/kg puerarin,2mg/kg astragaloside combined with 30mg/kg puerarin suspension intraperitoneal injection,continuous drug intervention for 4 days,can effectively protect liver injury in acute alcoholic rats.The mechanism of action may be to inhibit the mitochondria-enriched CYP2E1 protein expression and up-regulate the key protein Mortalin maintained by mitochondrial homeostasis,thereby alleviating the damage and damage of hepatocyte mitochondrial homeostasis by alcohol and its metabolites.3.Sesn family knockout mice can promote the development of nonalcoholic fatty liver induced by HFHFC diet.Its possible mechanism of action is that the Sesns knockout reduces the body's anti-inflammatory and anti-oxidative stress,increases oxidative stress and lipid metabolism,and leads to increased fatty liver.In addition,Sesns aggravates the development of inflammation and fibrosis by modulating the MAPK pathway to amplify hepatic stellate cells. |
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