| Objective To build an experimental model of Alcoholic fat liver disease in rats and evaluate the influence of Metadoxine in the process. To investigate the effects of Metadoxine on the expression levels of CYP 2E1 in liver tissues of alcoholic fatty liver rats.Methods Clean grade 30 male Wistar rats, weighing 200±10g, were randomly assigned into 3 groups(n=10).①alcoholic fatty liver group (fatty liver group), the rats were given 40%(v/v) dose-8g/(kg·d) ethanol, two times per day with an interval of 8-9h for 4 weeks. Since the fifth week would begin, the concentration of ethanol was increased to 50%(v/v), gavage to 8 weeks.②the prevention group (prevention group), the way of ethanol giver time interval and frequency was the same to the fatty liver group, while the ethanol was fed, Metadoxine, dose-300 mg/(kg·d) was given at the same time.③the control group (control group):given an equal volume of saline,the way of feeding was the same to the above two groups. In the end of 8 weeks, the model of alcoholic fat liver disease was established. Prepared the abdominal skin of all the rats, anesthetized the rats, opened the abdominal cavity, then the liver was excised for inspection, washed the blood, observed the general liver morphology, color, waiting for inspection. The tongue leaves of the liver was fixed by 4% formaldehyde solution, and stained with hematoxylin and eosin for light microscopic evaluation. Then, paraffin section was made for pathematology. Expression of mRNAs of CYP 2E1 were tested by RT-PCR, and immunohistochemistry. Futhermore, to investigate the effects of Metadoxine on the expression levels of CYP 2E1 in liver tissues of alcoholic fatty liver rats. Results In the process of establishing alcoholic fatty liver rat model, the alcoholic fatty liver group 2 died, both were caused by alcohol inhalation; the prevention group 2 died as a result of inhalation of the trachea, one died for the liver infection; the control group, no rat died.Imaging changes in the liver in control group by B-ultrasonic examination showed:the liver capsule is smooth, homogeneous echo, vascular textures clear. The fatty liver group 8 rats B-ultrasonic examination showed the liver echo slightly dense, slightly stronger, vascular texture is blurred; prevention group in rat liver 8 liver, B-under the echoes show a little dense, slightly stronger, vascular texture than the B group showed a clear. General and pathological changes in liver tissue in control rats the liver as a bright red, sharp edges, the surface smooth, no grainy aspect; fatty liver group, prevention group the liver were similar in general, have the volume increases, the aspect greasy. Liver, fatty liver group was the color brown-yellow color of the prevention group compared with the fatty liver group liver shallow, but lower than the control group deep. Observed under light microscope,①the fatty liver group:hepatocyte swelling, balloon-like change, point-like liver cell necrosis, was alcohol, body, slightly narrow sinusoidal liver cells, fatty degeneration of liver acinar zone 3 clearly shows periportal lymphoid, mononuclear cell infiltration;②prevention group:microscopic observation, and the fatty liver group compared with liver cell swelling abated, balloon-like fewer, alcohol, body less than the fatty liver group, the liver cells steatosis fatty liver group than light;③control group: rats with hepatic lobule integrity, a clear outline of the liver cells were polygonal cells around the central vein radial, sinusoid is clearly visible, liver cable arranged in neat rows, the basic structure of hepatic lobule portal area is clearly visible,. The difference was statistically significant ((P<0.05).Expression of mRNAs of CYP 2E1 were tested by immunohistochemistry.①fatty liver group:significantly increased expression of brown-yellow granules, and the distribution is a certain regularity, mostly concentrated in acinar zone 3 liver and portal vein to the 1,2 zone around the expansion of the distribution of fat change in line;②the preventive group, CYP 2E1 expression between the control, alcohol between the two groups;③the control group:CYP 2E1 expression is only found around the central vein area, was mainly expressed in hepatic acinar zone 3 liver cell membrane and plasma. Application of image proplus6.0 immunohistochemical image analysis system, gray value analysis, data analysis. The fatty liver group, prevention group in contrast with the control group, the expression of CYP 2E1 was significantly increased, (P<0.05); fatty liver group in contrast with the prevention group, CYP 2E1 expression increased, (P<0.05). By using the application Jetta 801 gel image analysis system, gray value analysis, data analysis, to investigate the expression of CYP 2E1 mRNA. The fatty liver group, prevention group in contrast with the control group, the expression of CYP 2E1 mRNA was significantly increased, (P<0.05); fatty liver group in contrast with the prevention group, CYP 2E1 mRNA expression increased, (P<0.05). (R=0.855, P<0.01).Conclusions By giving rats fed with a continuous alcohol, and using the way of testing pathological changes in liver tissue and liver imaging analysis showed that alcohol-induced fatty liver in this experimental rat model was successfully established.Conclusions The occurrence of alcoholic fatty liver increased with the proportion of CYP 2E1, and CYP 2E1 involved in the occurrence of alcoholic fatty liver and development. CYP 2E1 involved in the occurrence of alcoholic fatty liver and development of its expression increases with the incidence of alcoholic fatty liver is proportional to; Metadoxine CYP 2E1mRNA can effectively inhibit the expression of genes increases, reducing the expression level of CYP 2E1 influence of alcohol nature of the occurrence and development of fatty liver, thus effectively preventing the occurrence of nonalcoholic fatty liver, and the incidence of alcoholic fatty liver has a certain preventive effect.
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