TRANSGENIC RESCUE FROM FULMINANT HEPATITIS IN LEC RAT BY INTRODUCING HUMAN ATP7B cDNA / 解剖学报

ABSTRACT

Objective To investigate the possibility of affecting transgenic rescue for Wilson disease using the human ATP7B transgenic LEC rat.Methods The 7.1kb transgene constructed with human ATP7B cDNA and chicken ?-actin promoter was introduced into the fertilized oocytes of LEC rats, an animal model of Wilson disease, by microinjection. The expressions of human ATP7B protein in the transgenic rats were detected by Western blot. The plasma AST and ALT activities, and the total bilirubin levels in transgenic rats were measured continuously from 6 to 16 weeks using non-transgenic rats and LEA rat as controls. The pathological and histochemistry changes in the liver of the transgenic rats at 13 weeks were analyzed. Results The intact and correct product derived from human ATP7B was confirmed in the liver of transgenic rats. At the age around 12 weeks, the plasma AST and ALT activities, and the total bilirubin levels in transgenic rats were significantly decreased, while the inflammatory reation in the liver of transgenic rats was much mild as compared with that of non-transgenic rats, and the granules of stained copper were less in the hepatocytes of transgenic rats. By the age of 16 weeks, the transgenic rats were phenotypically normal, and the survival rate was 100%. These data showed that the LEC rats were successfully rescued from fulminant hepatitis after introducing of human ATP7B gene. Conclusion The hepatitis in Wilson disease is directly related to the toxicity of excessive accumulated copper, which attributed to the functional deficiency of the ATP7B. Gene transfer probably is the effective method for the therapy of Wilson disease.