| Objective Metadoxine (MTDX) is a new specific drug that is prescribed in healing alcoholic fatty liver disease in China. Though MTDX is widely used in clinic to treat alcoholic fatty liver, yet there are few studies put attention to its preventive effect. By analyzing the literatures, we found that CYP2E1 was closely related to alcoholic fatty liver disease, and there was no study had investigated the relation of MTDX and CYP2E1 in China. This paper tried to evaluate the preventive effect of MTDX in alcoholic fatty liver rat model of alcoholic fatty liver and explore its mechanism. Study the relationship between MTDX and CYP2E1, analyze whether CYP2E1 can become the MTDX therapeutic target.Method Clean grade 30 of Wistar male rats, weight (200±20)g, rats were given standard daily food and water in twice a day, and keep feeding the light and dark cycle 12 h 30 clean-level Wistar rats were randomly divided into three groups:control group, prevention group and model group. The alcoholic fat live was induced by alcohol (i.g.,8g·kg-1·d-1, twice daily for 8wk). Control group:rats were daily given the same volum of sterile water. Model group:1h after alcohol intragastric infusion, rats were given sterile water; Prevention group:1h after alcohol intragastric infusion, rats were given Metadoxine (300 mg·kg-·d-1). Ultrasound examination of rats to identified alcoholic fatty liver modeling situation, using automatic biochemical analyzer to detect the serum markers, H&E-stainning detect liver pathological changes, Oil red O staining detect liver fat deposition, Sirius red staining detect liver collagen, anthrone method for the detect of liver glycogen content, ELISA detect of lipoprotein Lipase, Determination of real-time quantitative PCR in liver tissue MIP-2, SOD-1, TGF-βgene expression, immunohistochemical staining detect of liver TNF-a, IL-6, CYP2E1 protein expression, Western blot assay CYP2E1 content in liver tissue. Used statistical analysis software SPSS 17.0.Results Alcoholic fatty liver rat model was successfully established, model group had one rat died due to alcohol misuse suction pipe, the other groups had no rats died.Model group and prevention group had lower body weight, compared with the control group was significant difference (P<0.05), revealing that the alcohol could affect the metabolism of rats. Liver of normal rats as bright red, sharp edges, smooth, virtually grain-free section; Model group could see the increased liver size, dark yellow, greasy slice of liver capsule smooth or jagged changes; Prevention group liver color lighter than model group, but slightly deeper than the control group, ultrasound imaging showed normal liver capsule was smooth, homogeneous echo, vascular texture clear and the liver size of normal. Model group liver ultrasound showed echogenic front, after the field echo attenuation, light intensity, vascular texture was unclear and fatty infiltration. Prevention group of rat liver was ultrasound examination showed slightly stronger than the echo, the distribution less uniform texture than the untreated group displayed blood vessels clear. H&E-stainning, Oil red O stainning and Sirius red staining showed that normal rats had pathological complete hepatic lobule, the liver cells were polygonal, radially around a central vein, sinusoids visible hepatic cord neatly arranged, the basic structure of hepatic lobule portal area is clearly visible, there is few fat liver cells deposition of collagen rare; model group liver cell was swelling, ballooning degeneration, lobular central deposition of fat droplets was very serious, it could be also seen that a lot of the red strip of collagen fibers, compared with the control group, ANOVA (P<0.01); prevention group:showed a small amount of fat deposition, portal area showed a small amount of collagen.Compared with the control group and prevention group, rats with alcoholic fatty liver model group, ALT, AST, TG was significantly higher (P<0.01, P<0.05), TC expression was low (P<0.01). Model group in liver glycogen content was significantly decreased compared with the control group (P<0.01); prevention of hepatic glycogen in rats was also lower than the control group (P<0.05), compared with model group, but there was no statistically significant. Liver fatty group was significantly higher lipoprotein lipase, single factor analysis of variance (P<0.01); prevention group glycogen content was higher than the control group (P<0.01), higher in the untreated group, but showed no statistical significance.After the induction of alcoholic fatty liver, TNF-a, IL-6 were significantly increased, consistent with previous studies reported, and MTDX effectively multi-symplectic TNF-a, IL-6 levels.CYP2E1 immunohistochemistry and western blot showed that the expression of the model group increased higher, compared with the control group, ANOVA (P<0.05); CYP2E1 expression in the prevention group significantly reduced compared with model group, the MTDX significantly inhibited the expression of CYP2E1.Conclusions Rats fed ethanol continuously to cause pathological changes used 8wk caused liver fat. Detect by ultrasound examination, pathology validate this model was successfully build alcoholic fatty liver disease model. Alcoholic fatty liver disease was closely related with the increase of CYP2E1. CYP2E1 and physiological factors could participate in the alcoholic fatty liver disease development. Prevention group decreased the expression of CYP2E1 protein levels, MTDX effectively inhibiting the expression of CYP2E1. CYP2E1 activity decreased could help patients with alcoholic liver disease liver function toward the virtuous circle of development. MTDX was effective in preventing the development of alcoholic fatty liver disease by down-regulating the expression of CYP2E1. |
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