Enzyme-Linked Immunosorbent Assay for Serum Procollagen Type III Peptide in Rats with Hepatic Fibrosis.

Abstract

ABSTRACT. The process of hepatic fibrosis, and the changes in contents of hepatic hyproxyproline (HYP) and serum procollagen type III peptide (PIIINP) were examined in two rat models for hepatic fibrosis, i.e. bile duct ligation/scission (BDL/s)-and dimethylnitrosamine (DMN)-induced models. In addition, an expression of type III collagen mRNA in the liver of BDL/s model was also examined. In BDL/ s model, hepatic fibrosis started at 2 weeks after operation (WAO) and cirrhosis with prominent bile duct hyperplasia was detected at and after 5 WAO. Serum PIIINP content measured using a modified double armed inhibition enzyme-linked immunosorbent assay (ELISA) method proposed by us started to increase at 1 WAO and continued to increase thereafter. Hepatic HYP content measured colorimetrically started to increase at 3 WAO and it continued to increase until 7 WAO. An expression of type III collagen mRNA in the liver was enhanced at and after 2 WAO, especially at 4 and 5 WAO. In DMN model, marked hepatic fibrosis was detected at 1 week after the last DMN administration (WAA), and the degree of fibrosis was apparently reduced at 4 WAA. Serum PIIINP content prominently increased at 1 WAA and decreased at and after 3 WAA. Hepatic HYP content showed a marked increase at 1 WAA and decreased thereafter. The present results indicated that the sequences of hepatic fibrosis, hepatic HYP content and serum PIIINP content were well correlated with each other in both BDL/s and DMN models. In conclusion, ELISA system for the detection of serum PIIINP content is considered to be reliable method for assessment of cirrhotic liver, and the present two rat models for liver fibrosis/cirrhosis seems to be a good tool for researching antifibrotic agents. -KEY WORDS: enzyme-linked immunosorbent assay, hepatic fibrosis model, rat, serum procollagen type III peptide.J. Vet. Med. Sci. 60(11): 1213-1220, 1998 and hepatocyte necrosis. Namely, bile duct ligation/scission (BDL/s) is recommended as a useful method for producing progressive and reproducible biliary fibrosis in rats without prominent hepatocyte necrosis and severe inflammation [9,19]. In addition, treatment with dimethylnitrosamine (DMN) is also known to have an advantage to induce fibrosis in rats in a relatively short period (3 weeks) with slight inflammation and hepatocyte necrosis [2,26]. The aim of this study is to establish a non-radioactive method, i.e. enzyme-linked immunosorbent assay (ELISA) to evaluate serum PIIINP using BDL/s-and DMN-induced hepatic fibrosis in rats. MATERIALS AND METHODS Animals:Two hundred male Sprague-Dawley rats (11 weeks old) obtained from Laboratory Animal Center of Seoul National University in Korea were used. The animals were kept in an animal room maintained at 23 ± 2°C with 60 ± 10% r.h. with 12 hr-light and 12 hr-dark cycle, and were given normal pelleted diet (Shinchon, Korea) and water ad libitum.Treatments: As to bile duct ligation/scission (BDL/s) model, the hepatic biliary system was occluded by distal and proximal ligation using 4-0 silk (P...

Protein fingerprinting of the extracellular matrix remodelling in a rat model of liver fibrosis—a serological evaluation

“…All formation markers were elevated in the first quartile of total hepatic collagen indicating that serum levels of collagen formation fragments may be valuable markers of initial fibrosis formation and thus early diagnosis. These data are in alignment with previous studies showing that the formation of type III (PIIINP) and type IV (PIVNP) collagen are related to fibrosis . Contradictory data have been reported for the serum assessment of intact type VI collagen in relation to its correlation with liver fibrosis.…”