Screening Affinity Short Peptide Of TβRⅡ And Studying On Its Anti-hepatic Fibrosis Effect In Mice

Objective: To screen high affinity short peptides of type II transforming growth factor beta receptor from phage displayed 12-mer peptide library, and to study the effects of affinity short peptides on anti-hepatic fibrosis in mice.Methods: In this study, recombinant solube human type II transforming growth factor beta receptor was used as the target to screen its short affinity peptide from phage displayed 12-mer peptide library. The positive phage clones were screened out by three rounds of biopanning, followed by single stranded DNA extraction and sequence analysis. The affinity of high frequent short peptide was identied by ELISA, and applied to the antifibrotic study. Sixty male BALB/c mice were divided randomly into six groups: 10 in experimental liver fibrotic model induced by carbon tetrachloride as disease control group; 10 in short peptide C1 as treatment group one; 10 in short peptide C2 as treatment group two; 10 in short peptide C3 as treatment group three; 10 in colchicine as treatment control group and 10 in normal control group. All of them were sacrificed after five weeks. The serum levels of ALT,ALB,TP and the contents of hepatic hydroxyproline were determined by biochemistry method. Liver tissue was handled with HE and Masson staining for hispathological study.Results: The screening experiment obtained six kinds of nucleotide sequence from twelve positive phage clones combining type II transforming growth factor beta receptor with affinity. There were not conservative sequence and homologous sequence compared with transfoming growth factor beta 1 through sequence analysis. Affinity short peptide C2 and C3 didn't showed high affinity to combine with type II transforming growth factor beta receptor and any obvious effect on anti-hepatic fibrosis. C1 showed high affinity to combine with type II transforming growth factor beta receptor. The inflamation weakened, and collagen fiber synthesis decreased significantly in C1 treatment group. The hepatic fibrosis degree reduced obviously, of which the difference was statistically significant compared with the fibrotic mice model group (P<0.05).Conclusions: The study showed that affinity short peptide can be screened from phage displayed 12-mer peptide library. Liver fibrotic model was successfully induced by carbon tetrachloride. Affinity short peptide C1 had effect on preventing hepatic fibrosis. It might provide a new way to anti-hepatic fibrosis.