Molecular cloning, characterization and mRNA expression of six peroxiredoxins from Black carp Mylopharyngodon piceus in response to lipopolysaccharide challenge or dietary carbohydrate.

Peroxiredoxin (Prx) belongs to a cellular antioxidant protein family that plays important roles in innate immune function and anti-oxidative capability. In the present study, six Prxs were cloned from Black carp Mylopharyngodon piceus (MpPrx) by homology cloning and rapid amplification of cDNA ends (RACE) techniques. There were 199, 197, 250, 260, 189 and 222 amino acids in six MpPrxs, respectively. BLAST analysis reveals that MpPrxs shares high identities and similar characteristics with other known Prxs from animals. The phylogenetic analysis evidenced three major subclasses corresponding to one-Cys-Prx (MpPrx6), typical two-Cys-Prx (MpPrx1-4) and atypical 2-Cys-Prx (MpPrx5) that reflected the present hierarchy of vertebrates and invertebrates. Although six MpPrxs are constitutively expressed in all tissues, relatively higher-level mRNA expression levels of six MpPrxs can be detected in liver, eyes, heart and adipose tissues by real-time PCR assays. The transcriptional patterns of six MpPrxs mRNA in liver were detected by real-time PCR in Black carp after lipopolysaccharide (LPS) challenge and treated with graded levels of dietary carbohydrate (CHO) (106.5, 194.3, 288.4 and 379.1 g kg(-1)), respectively. These results showed that stimulation with LPS could induce up-expression of six MpPrxs mRNA, and the variations of MpPrx4 were more sensitive than these of other MpPrxs in the liver of Black carp. Compared with those in group with 106.5 g kg(-1) dietary CHO, the expression levels of MpPrx2, MpPrx3 and MpPrx6 were significantly down-regulated while MpPrx5 were significantly induced in liver of Black carp fed with adequate dietary CHO (194.3 g kg(-1)). In addition, significant up-regulations of MpPrx2, MpPrx3 and MpPrx6 were observed in Black carp fed with excessive dietary CHO (379.1 g kg(-1)). And MpPrx4 could be constantly induced with increasing dietary CHO contents in this study. These results indicated that MpPrxs were constitutive and inducible proteins and might play important roles in innate immune function after LPS challenge and regulating redox homeostasis in the metabolism of dietary CHO.