Synergistic liver toxicity of copper and retrorsine in the rat

To investigate the possible synergy between copper and retrorsine (a pyrrolizidine alkaloid) as a cause of Indian Childhood Cirrhosis, four groups of male Wistar rats were fed the following diets from weaning: A. Normal diet; B. Copper loaded (2 g CuSO4/kg diet); C. Retrorsine supplemented (Expt 1:25 mg/kg body weight/week by gavage, Expt 2:25 mg/kg food initially then 15 mg/kg food after 4 weeks); and D. Copper and retrorsine as above. Serial plasma samples were assayed for aminotransferases, albumin and bilirubin. Liver samples at biopsy and sacrifice provided samples for copper analysis and histology. Results showed that copper and retrorsine together significantly increased liver damage compared with feeding either alone as assessed by: 1. Increased mortality rate; 2. Decreased plasma albumin and increased plasma bilirubin (mainly conjugated) indicative of hepatocyte dysfunction; 3. Massive liver copper accumulation, and 4. Increased liver damage histologically. Thus retrorsine caused liver copper accumulation, and together copper and retrorsine led to severe hepatic dysfunction, characterised by hypoalbuminaemia and conjugated hyperbilirubinaemia. Plant alkaloids secreted in milk by grazing animals and copper from brass vessels may together produce Indian Childhood Cirrhosis.