Wu Chenglong, Gao Jun'e, Cao Fang, Lu Zhibin, Chen Lian, Ye Jinyun
School of Life Science, Huzhou University, 759 Erhuan Road (E), Huzhou, 313000, PR China.
School of Life Science, Huzhou University, 759 Erhuan Road (E), Huzhou, 313000, PR China.
Fish Shellfish Immunol. 2016 Mar;50:210-22. doi: 10.1016/j.fsi.2016.01.033. Epub 2016 Jan 30.
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.
过氧化物酶（Prx）属于细胞抗氧化蛋白家族，在先天免疫功能和抗氧化能力中发挥重要作用。在本研究中，通过同源克隆和cDNA末端快速扩增（RACE）技术从草鱼（Mylopharyngodon piceus，MpPrx）中克隆出6种Prx。6种MpPrx分别含有199、197、250、260、189和222个氨基酸。BLAST分析表明，MpPrx与其他已知动物Prx具有高度同源性和相似特征。系统发育分析证明了三个主要亚类，分别对应单半胱氨酸Prx（MpPrx6）、典型双半胱氨酸Prx（MpPrx1 - 4）和非典型双半胱氨酸Prx（MpPrx5），这反映了脊椎动物和无脊椎动物目前的层级关系。尽管6种MpPrx在所有组织中组成性表达，但通过实时PCR检测发现，在肝脏、眼睛、心脏和脂肪组织中可检测到相对较高水平的6种MpPrx的mRNA表达。分别用脂多糖（LPS）刺激草鱼，并给予不同水平的饲料碳水化合物（CHO）（106.5、194.3、288.4和379.1 g kg(-1)）处理后，通过实时PCR检测肝脏中6种MpPrx mRNA的转录模式。结果表明，LPS刺激可诱导6种MpPrx mRNA的上调表达，且在草鱼肝脏中，MpPrx4的变化比其他MpPrx更敏感。与饲料CHO含量为106.5 g kg(-1)组相比，在饲料CHO含量充足（194.3 g kg(-1)）的草鱼肝脏中，MpPrx2、MpPrx3和MpPrx6的表达水平显著下调，而MpPrx5显著上调。此外，在饲料CHO含量过高（379.1 g kg(-1)）的草鱼中，观察到MpPrx2、MpPrx3和MpPrx6显著上调。并且在本研究中，随着饲料CHO含量的增加，MpPrx4可不断被诱导。这些结果表明，MpPrx是组成性和可诱导性蛋白，可能在LPS刺激后的先天免疫功能以及调节饲料CHO代谢中的氧化还原稳态中发挥重要作用。
