动物造模 z-bo 2020-08-07 500

　　(1) Replication method Adult rats, after normal feeding for 1 week, change to a high-fat diet (add 1% cholesterol and 5% lard on the basis of a low-fat diet), and at the same time give 60% ethanol 1.5ml/kg body weight Stomach, twice a day, for 12 weeks, fasting for 16 hours at the end of the experiment, blood was taken to prepare serum, liver tissue was taken to prepare homogenate, and paraffin sections were used for biochemical analysis and histomorphological examination. Or while feeding a high-fat diet, use 60% ethanol 15ml/kg body weight gavage twice a day for 3-6 months. Or at the same time of regular feeding, daily infusion of liquor, olive oil, and pyrazole mixture, in which the intake of liquor is 8-12g/kg body weight, and increases with time, that is, 8g in the first week. 10g in the second week, 12g from the third week, and continue to maintain this amount until the end of the experiment. The intake of pyrazole is 27.2mg/kg body weight daily, once a day for 4 consecutive weeks, after the last dosing Samples were taken at 24h, and embedded in conventional paraffin wax and electron microscope for histological observation.

　　(2) Model features High-fat diet plus 60% ethanol 1.5ml/kg body weight gavage for 12 weeks, model rat serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), cholinesterase (ChE) , Triacylglycerol (TG), malondialdehyde (MDA) and liver tissue MDA increased, serum glutathione peroxidase (GSH-Px) and liver tissue superoxide dismutase (SOD), glutathione Peptide (GSH) decreased; under the microscope, hepatocytes were obviously swollen, the cytoplasm was loose, and the balloon was changed. In the cytoplasm, there were also lipid droplet vacuoles of different sizes and numbers, and Mallory (alcoholic hyalosis) was also seen. Body and focal necrosis, inflammatory cell infiltration in the interstitium. High-fat diet plus 60% ethanol 15ml/kg body weight gavage for 3 months, model animals showed mild to moderate steatosis, the number of fat storage cells and liver tissue a-smooth actin (a-SMA) positive cells increased, 6 At the time of month, fatty degeneration of liver cells worsened, a-SMA positive cells increased, and fibrosis around central vein and perisinus could be seen. Routine diet with white wine, olive oil, and pyrazole gavage for 4 weeks. Under the microscope, it can be seen that the central area of the liver lobules of the model animals is obviously necrosis, or focal necrosis, or punctate necrosis, with a large number of inflammatory cell infiltration in the interstitium; electron microscopy shows that its liver The nucleus is irregular, and the perinuclear space is expanded and uneven; the cytoplasm is loose, the electron density is reduced, and the number of organelles is reduced; the mitochondria are swollen and deformed, the cristae disappear, and the number of abnormal mitochondria is large; the rough endoplasmic reticulum expands, breaks, and Degranulation, mostly large and small vesicles; lipid droplet vacuoles of different sizes and numbers, and alcoholic bodies arranged in filaments are also seen in the cytoplasm; lipid storage cells are active and unique in them The lipid droplet vacuoles are reduced or disappeared; proliferating collagen fibers can be seen in the perisinus space and around the liver cells.

　　(3) Comparative Medicine Alcoholic liver disease includes alcoholic fatty liver, alcoholic hepatitis, alcoholic liver fibrosis, and alcoholic cirrhosis. These lesions are often progressive and can partially overlap. After ethanol is absorbed from the gastrointestinal tract, the vast majority (greater than 90%) is oxidized in the liver by alcohol dehydrogenase (ADH) and some microsomal enzymes (microsomal alcohol oxidation system) and then metabolized. The main metabolite of ethanol is acetaldehyde, which has toxic effects on the liver and other organs. Clinically, different biochemical indicators can comprehensively reflect the degree of liver cell damage (ALT, AST), reserve capacity (ChE) and metabolic function (TBIL, TP, ALB, TG) from different aspects, and the histomorphological examination is in To a certain extent, it is the most direct evidence to distinguish alcoholic liver disease and its different stages. Mallory corpuscles are caused by some cytoplasmic fibrin deposition in swollen liver cells, which contain little or no fat, and are specific lesions of alcoholic liver disease. In addition, the connective tissue around the liver sinusoids and hepatocytes can be seen in the central zone of the liver acinar, and the collagen fibers form a continuous membrane-like structure under the sinusoidal endothelium. Alcoholic hepatitis with diffuse inflammatory cell infiltration and necrosis can be seen in the intermediate process of fatty liver developing into cirrhosis. Inflammatory cell infiltration and fatty liver are its characteristic lesions. The biochemical indicators and liver tissue morphological examination of the above-mentioned animal models are basically consistent with the characteristics of human alcoholic liver disease, and they are currently the main modeling methods used in China. Among them, the high-fat diet plus 60% ethanol 1.5ml/kg body weight gavage for 12 weeks is similar to the fatty liver disease of alcoholic liver disease, while the high-fat diet plus 60% ethanol 15ml/kg body weight gavage 6 months model or conventional diet The model with white wine, olive oil and pyrazole gavage for 4 weeks is more similar to the pathological changes of alcoholic liver fibrosis. However, for the alcoholic liver fibrosis model that is replicated by alcohol gavage, the former has a low formation rate and atypical lesions.