Pathological Model Construction Of Cytochrome P4502E1 Regulating Fatty Liver Caused By Alcohol And Hyperlipidemia

Objective:(1)to establish an animal model of obesity and alcohol synergistic steatohepatitis in cytochrome P4502E1(CYP2E1)knockout mice.(2)to investigate the role of CYP2E1 in the synergistic effects of alcohol and high fat steatohepatitis.Methods: firstly,the CYP2E1 gene knockout mice were bred and bred,and the homozygote was screened by tail gene identification.Then the 7 week old male CYP2E1 knockout mice and wild type mice were also divided into 4 groups respectively: 1)normal diet(control)group: gastrostomy,through micro injection pump daily with 2 l/min velocity,liquid feed 580kcal/kg/d routine,constant speed infusion of small rats;2)alcohol group: gastrostomy,through micro injection pump daily at a flow rate of 2ul/min,580kcal/kg/d and 15g/kg/d of the conventional feed liquid alcohol,constant infusion of mice;3)high fat high cholesterol group: gastrostomy,through micro injection pump daily with 2 l/ min velocity.Liquid 780kcal/kg/d(high fat diet containing 0.21% cholesterol)constant infusion of mice;4)alcohol together with high fat and high cholesterol group: gastrostomy,through micro injection pump daily with 2 l/min liquid flow rate,high fat diet 780kcal/kg/d(containing 0.21% cholesterol)and 15g/kg/d alcohol,constant infusion of mice.After 2 weeks,the frozen sections of liver were collected and analyzed by oil red O and routine HE staining.Results: the liver tissues were collected for pathological analysis.It was found that the liver cells showed ballooning degeneration and occasionally spot necrosis.Large amounts of fat droplets were seen in oil red O staining.At the same time,the liver tissue was detected by Real-Time and qPCR,and it was found that alcohol and high fat can promote the increase of CYP2E1 expression and promote the expression of inflammatory factors,which indicated that the obese alcohol synergistic fatty liver model was successful.Conclusion:(1)a hepatitis model with high lipid and alcohol synergistic effects in mice was successfully constructed.(2)CYP2E1 was found to mediate inflammation under the combined effects of high fat,alcohol,and high fat alcohol.