Thioredoxins (Trxs) are a family of small and highly conserved proteins which play crucial roles in the maintenance and regulation of the cellular redox homeostasis. In this study, the full-length cDNAs of thioredoxin 1 (TfTrx1) and thioredoxin-related protein of 14?kDa (TfTrp14) were isolated from roughskin sculpin (Trachidermus fasciatus). TfTrx1 is 662?bp in length with a 336-bp open reading frame (ORF) that encodes for a peptide with 111 amino acids, and TfTrp14 consists of 1066?bp with a 372-bp ORF that is translated to 123 amino acids. TfTrx1 and TfTrp14 contain highly conserved catalytic site motif CGPC and CPDC, respectively. Tissue distribution analysis indicated that both genes were broadly expressed in all examined tissues with the highest expression of TfTrx1 in the blood and TfTrp14 in the brain. In post-LPS and heavy metal challenge, the mRNA of both genes was significantly increased in the skin, liver, spleen, and brain at various times. The results of western blot detection displayed that the time of the induced maximum protein expression was 6-h post-LPS injection in the skin and liver, which were slightly delayed compared with that of 2?h at mRNA level. The recombinant TfTrp14 and TfTrx1 proteins were expressed in E. coli BL21 (DE3). The increase of the fluorescence intensity in rTfTrx1 and rTfTrp14 suggested the redox state changes in the microenvironment around tryptophan residues. Both of the recombinant proteins exhibited concentration-dependent disulfide reductase activity towards insulin, and the catalytic activity of rTfTrx1 was much higher than that of rTfTrp14.