Molecular characterization, expression analysis and function identification of Pf TNF-alpha and its two receptors Pf TNFR1 and Pf TNFR2 in yellow catfish (Pelteobagrus fulvidraco)
Molecular characterization, expression analysis and function identification of Pf TNF-alpha and its two receptors Pf TNFR1 and Pf TNFR2 in yellow catfish (Pelteobagrus fulvidraco)
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黄颡鱼Pf TNF-α及其两个受体Pf TNFR1和Pf TNFR2的分子表征、表达分析及功能鉴定
DOI：
10.1016/j.ijbiomac.2021.06.09
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发表时间：
2021
影响因子：
8.2
通讯作者：
Wei Kai-Jian
中科院分区：
文献类型：
--
作者：
Hu Yuan-Hong;Zhou Xu;Jiang Xin-Xin;Zhang Gui-Rong;Shi Ze-Chao;Ji Wei;Ma Xu-Fa;Luo Zun-Lan;Wei Kai-Jian
炎症是机体免疫的常见表现，并介导细胞因子的级联反应。肿瘤坏死因子-α（TNF-α）作为一种多效细胞因子，通过与其受体（TNFR）相互作用在炎症反应中发挥重要作用。本研究克隆了黄颡鱼（Pelteobagrus fulvidraco）的Pf_TNF-α、Pf_TNFR1和Pf_TNFR2基因，生物信息学分析表明，这3个基因具有与其他脊椎动物相似的保守序列特征。qPCR结果显示，Pf_TNF-α、Pf_TNFR1和Pf_TNFR2 mRNA在14种组织和4种组织的淋巴细胞中呈组成型表达。感染后，脾脏、肝脏、干肾、头肾和鳃中Pf_TNF-α和Pf_TNFR1基因的mRNA表达水平明显上调，而Pf_TNFR2基因的mRNA表达水平在脾脏中明显上调，在肝脏和鳃中下调。在黄颡鱼外周血白细胞（PBL）中，经脂多糖（LPS）、肽聚糖（PGN）、多聚肌苷酸-多聚胞苷酸（Poly I：C）和植物血凝素（PHA）刺激后，Pf_TNF-α mRNA表达显著上调，Pf_TNFR转录本表达显著下调。经重组（r）Pf_sTNF蛋白刺激后，PBLs中多种炎症因子基因的mRNA表达上调。rPf_sTNF可促进白细胞的吞噬活性，而rPf_TNFR1CRD 2/3和rPf_TNFR2CRD 2/3可抑制rPf_sTNF介导的吞噬活性。MAPK抑制剂VX-702和NF-κB抑制剂PDTC可抑制rPf_sTNF诱导的TNF-α和IL-1β mRNA表达上调。rPf_sTNF诱导PBLs表达FADD mRNA，增加PBLs凋亡率，抑制NF-κB和MAPK信号通路可促进PBLs凋亡。结果表明，Pf_TNF-α、Pf_TNFR1和Pf_TNFR2在黄颡鱼抵抗细菌入侵的免疫应答中起重要作用。
Inflammation is a common manifestation of body immunity and mediates a cascade of cytokines. Tumor necrosis factor-α (TNF-α), as a multi-effect cytokine, plays an important role in the inflammatory response by interacting with its receptor (TNFR). In this study,Pf_TNF-α,Pf_TNFR1 andPf_TNFR2 genes were cloned from yellow catfish (Pelteobagrus fulvidraco), and bioinformatics analyses showed that the three genes were conserved and possessed similar sequence characteristics as those of other vertebrates. The qPCR results showed thatPf_TNF-α,Pf_TNFR1 andPf_TNFR2 mRNAs were constitutively expressed in 14 tissues and the lymphocytes of four tissues from healthy adults. The mRNA expression levels ofPf_TNF-α andPf_TNFR1 genes were significantly up-regulated in the spleen, liver, trunk kidney, head kidney and gill afterEdwardsiella ictaluriinfection, while the mRNA expression ofPf_TNFR2 was significantly up-regulated in the spleen, and down-regulated in the liver and gill. In the isolated peripheral blood leukocytes (PBLs) of yellow catfish, the expression ofPf_TNF-α mRNA was notably up-regulated and the twoPf_TNFR transcripts were distinctly down-regulated after stimulation with lipopolysaccharides (LPS), peptidoglycan (PGN), polyinosinic-polycytidylic acid (Poly I:C) and phytohaemagglutinin (PHA). After stimulated by recombinant (r)Pf_sTNF protein, the mRNA expressions of various inflammatory factors genes were up-regulated in the PBLs. Meanwhile, rPf_sTNF promoted the phagocytic activity of leukocytes, whereas the activity mediated by rPf_sTNF could be inhibited by rPf_TNFR1CRD2/3 and rPf_TNFR2CRD2/3. The up-regulation of TNF-α and IL-1β mRNAs expression triggered by rPf_sTNF could be inhibited by MAPK inhibitor (VX-702) and NF-κB inhibitor (PDTC). rPf_sTNF induced the expression of FADD mRNA in PBLs and increased the apoptotic rate of PBLs, and inhibiting the NF-κB and MAPK signal pathways could enhance the apoptosis of PBLs. The results indicate thatPf_TNF-α,Pf_TNFR1 andPf_TNFR2 play important roles in the immune response of yellow catfish to bacterial invasion.