Abstract
Intestinal trefoil factor (ITF) is a small polypeptide with potential medical values whose main pharmacological effects are to alleviate gastrointestinal mucosal injury caused by various injury factors and promote the repair of damaged mucosa. However, its low yield limits its application. The purpose of our study was to construct a recombinant adenoviral vector containing the hITF gene and observe the therapeutic effect of burn-induced intestinal mucosal injury using in vitro and in vivo analysis. First, a recombinant shuttle plasmid was constructed by ligating a pAdTrack-CMV vector with a full-length hITF gene containing a signal peptide and the mature peptide, followed by the recombinant Ad-hITF adenovirus vector after linearization and homologous recombination with the backbone plasmid in the competent BJ5183 strain. Second, the hITF expression level was detected using reverse transcription polymerase chain reaction and western blotting after Ad-hITF infection of colon cancer HT-29 cells. The recombinant adenovirus significantly promoted cell migration in an in vitro wounding model. Finally, we confirmed that the recombinant adenovirus could significantly expedite the healing of intestinal mucosal injury after establishing a mouse model in which severe burns were stimulated and the recombinant adenovirus was delivered by intragastric injection. In summary, we constructed a recombinant adenoviral vector containing the hITF gene and confirmed its role in promoting repair of the intestinal mucosa. Our study provides a novel way to treat burn-induced intestinal mucosal injury.
Highlights
Intestinal trefoil factor (ITF), a small polypeptide secreted by intestinal goblet cells [1,2], was named for its characteristic ‘‘trefoil’’ disulphide loop composed of a special core domain in its 59-amino-acid sequence
ITF, a small polypeptide with potential medical value, alleviates the gastrointestinal mucosal injury caused by various injury factors and promotes the repair of damaged mucosa
Full-length hITF cDNA contains 222-bp DNA (GenBank Accession No NM003226) that encodes 73 amino acids including the first 14aa signal peptide followed by a 59-aa mature peptide [13,14]
Summary
Intestinal trefoil factor (ITF), a small polypeptide secreted by intestinal goblet cells [1,2], was named for its characteristic ‘‘trefoil’’ disulphide loop composed of a special core domain in its 59-amino-acid sequence. The core domain contains 6 cysteine residues linked by disulfide bonds in specific positions and forms 3 cyclic structures that comprise a trefoil-like structure after bending and folding of the entire peptide chain [3]. A newly emerged means of treatment, treats diseases by delivering normal human gene or gene with a therapeutic effect into human target cells in a certain way to correct genetic defects or play a therapeutic role [6,7]. If ITF can be delivered into human body by gene therapy and abundantly expressed in vivo, it will both simplify the tedious preparation procedure and reduce production costs
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