| 摘要: |
| 由细菌等感染引起的疾病对中华乌塘鳢(Bostrychus sinensis)养殖业的可持续发展造成严重威胁。白细胞介素8 (Interleukin-8, IL-8)在调节炎症反应和抗感染免疫方面扮演着重要的作用。利用PCR扩增和克隆测序获得了中华乌塘鳢IL-8 (命名为BsIL-8) cDNA序列。BsIL-8 cDNA序列全长1 244 bp, 包含576 bp的5’端非编码区序列(5’-UTR), 312 bp的开放阅读框序列(ORF)和356 bp的3’端非编码区序列(3’-UTR)。BsIL-8共编码103个氨基酸, 包含1个由18个氨基酸组成的信号肽和1个由62个氨基酸组成的SCY结构域。其中, SCY结构域包含1个CXC基序(Cys30-Arg31-Cys32)以及4个保守的半胱氨酸残基(Cys30、Cys32、Cys57和Cys73)。序列分析结果表明, BsIL-8与大弹涂鱼(Boleophthalmus pectinirostris)的IL-8序列相似性和一致性最高(分别为92.4%和86.4%)。基因组织表达分布结果显示, BsIL-8基因在主要组织器官中呈泛在性表达, 头肾BsIL-8基础表达量最高, 肝脏次之, 外周血最低。Poly (I:C)免疫刺激后, BsIL-8在4个重要免疫器官(脾脏、肝脏、头肾和外周血)中的表达量均发生了显著的上调(分别在刺激后3、12、24和24 h时表达量达到最高, P<0.01)。副溶血弧菌(Vibrio parahaemolyticus)感染后, BsIL-8基因在脾脏和外周血中的表达量均显著上调(均在感染后12 h时表达量达到最高, P<0.01), 而在肝脏和头肾中的表达量均显著下调(均在感染后12 h时表达量达到最低, P<0.01)。综上所述, BsIL-8基因参与了中华乌塘鳢抗感染免疫应答过程。 |
| 关键词: 中华乌塘鳢(Bostrychus sinensis) 白细胞介素8 (IL-8) 基因表达 免疫应答 副溶血弧菌 |
| DOI：10.11693/hyhz20231200278 |
| 分类号:S941;S965;Q789 |
| 基金项目:国家自然科学基金项目,42106101号。 |
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| MOLECULAR CLONING AND EXPRESSION ANALYSES OF THE INTERLEUKIN-8 GENE FROM BOSTRYCHUS SINENSIS IN RESPONSE TO IMMUNE STIMULATION |
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CHEN Lin-Jie, SHEN Bin, ZHANG Jian-She
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National Engineering Research Center for Marine Aquaculture, Zhejiang Ocean University, Zhoushan 316022, China
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| Abstract: |
| The sustainable development of Bostrychus sinensis aquaculture has been seriously impacted by bacterial infectious diseases. Interleukin-8 (IL-8) plays an important role in anti-infective immunity and regulation of inflammatory responses. The cDNA sequence of B. sinensis IL-8 (named BsIL-8) was obtained by PCR amplification and clonal sequencing. The cDNA sequence of BsIL-8 is 1 244 bp in length, comprising a 5’-untranslated region (5’-UTR) of 576 bp, an open reading frame (ORF) of 312 bp, and a 3'-UTR of 356 bp. BsIL-8 encodes 103 amino acids, including an 18-amino acid signal peptide and a 62-amino acid SCY domain. The SCY domain contains a CXC motif (Cys30-Arg31-Cys32) and four conserved cysteine residues (Cys30, Cys32, Cys57, and Cys73). Sequence analyses showed that BsIL-8 had the highest sequence similarity and identity with Boleophthalmus pectinirostris IL-8 (92.4% and 86.4%, respectively). The RT-PCR results revealed that BsIL-8 was constitutively expressed in the 10 organs/tissues examined in B. sinensis. The head kidney exhibited the highest expression, followed by the liver, while the peripheral blood showed the lowest expression. Following immune stimulation with poly (I︰C), the expression of the BsIL-8 gene was significantly upregulated in the spleen, liver, head kidney, and peripheral blood of B. sinensis. The highest expression levels were observed at 3, 12, 24, and 24 h post-stimulation, respectively (P<0.01). After Vibrio parahaemolyticus infection, the expression of the BsIL-8 gene was significantly upregulated in the spleen and peripheral blood of B. sinensis, with the highest expression levels observed at 12 h post-infection (P<0.01). Conversely, the liver and head kidney exhibited significant downregulation of the gene, with the lowest expression levels detected at 12 h post-infection (P<0.01). In conclusion, the BsIL-8 gene is involved in the anti-infection immune responses of B. sinensis. |
| Key words: Bostrychus sinensis IL-8 (Interleukin-8) gene expression immune response Vibrio parahaemolyticus |
