Molecular characterization, expression analysis, and bactericidal activity of the derivative peptides of TFPI-1 and TFPI-2 in half-smooth tongue sole, Cynoglossus semilaevis.
Molecular characterization, expression analysis, and bactericidal activity of the derivative peptides of TFPI-1 and TFPI-2 in half-smooth tongue sole, Cynoglossus semilaevis.
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DOI：
10.1016/j.fsi.2016.10.003
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发表时间：
2016-11
影响因子：
4.7
通讯作者：
Xin-peng Zhao;Shu-Wen He;B. Yue;Guang-hua Wang;M. Zhang
中科院分区：
文献类型：
--
作者：
Xin-peng Zhao;Shu-Wen He;B. Yue;Guang-hua Wang;M. Zhang
组织因子途径抑制剂（Tissue factor pathway inhibitors, tfpi）是一种可逆性调节组织因子诱导的凝血的kuniz型丝氨酸蛋白酶抑制剂。TFPI族包含两个成员，TFPI-1和TFPI-2。近年来的研究表明，TFPI-1和TFPI-2在先天免疫中也发挥着重要作用，然而，硬骨鱼TFPI的潜在功能非常有限。本研究对半光滑舌鳎（Cynoglossus semilaevis）中两个TFPI分子（CsTFPI-1和CsTFPI-2）进行了表征，研究了它们在病原体刺激下的组织分布和表达模式，并研究了c端肽的抗菌活性。实时定量RT-PCR分析显示，在头肾、肠、脑、脾、肝、皮肤、鳃、心脏和肌肉中均有组成型的表达，表达量依次递增；cstfpi -2在鳃、肠、皮肤、头肾、肝、脑、脾、肌肉和心脏中依次表达。在鳗弧菌、无乳链球菌和鱼巨细胞病毒刺激下，CsTFPI-1和CsTFPI-2的表达随病原菌、组织类型和感染阶段的不同而显著升高，提示CsTFPI-1和CsTFPI-2在抗菌和抗病毒感染中发挥重要作用。最后合成了CsTFPI-1和CsTFPI-2的c端肽，证明其对黄体微球菌具有不依赖宿主血清的抗菌作用。综上所述，这些结果表明CsTFPI-1和CsTFPI-2在该鱼的抗菌免疫中起重要作用。
Tissue factor pathway inhibitors (TFPIs) are Kunitz-type serine protease inhibitors that reversibly regulate the blood coagulation induced by tissue factor. TFPI family contain two members, TFPI-1 and TFPI-2. Recent studies have shown TFPI-1 and TFPI-2 also play important roles in innate immunity, however, the potential function of teleost TFPI are very limited. In this study, we characterized two TFPI (CsTFPI-1 and CsTFPI-2) molecules from half-smooth tongue sole (Cynoglossus semilaevis), examined their tissue distributions and expression patterns under pathogens stimulation as well as investigated the antibacterial activity of the C-terminal peptides. Quantitative real time RT-PCR analysis showed that constitutiveCsTFPI-1expression occurred, in increasing order, in head kidney, intestine, brain, spleen, liver, skin, gills, heart, and muscle;CsTFPI-2was expressed, in increasing order, in the gills, intestine, skin, head kidney, liver, brain, spleen, muscle, and heart. Under Vibrio anguillarum, Streptococcus agalactiae and fish megalocytivirus stimulation, bothCsTFPI-1andCsTFPI-2expression increased significantly in a manner that depended on the pathogen, tissue type, and infection stage, which suggested CsTFPI-1 and CsTFPI-2 play important roles in anti-bacterial and anti-viral infection. Finally, C-terminal peptides of CsTFPI-1 and CsTFPI-2, were synthesized and proved to have antibacterial effect against Micrococcus luteus that were independent of host serum. Take together, these results indicate that CsTFPI-1 and CsTFPI-2 play important roles in antimicrobial immunity of this fish.