TRAF2在肉鸡肺动脉高压中的潜在作用及其多克隆抗体的制备和特异性分析。

IF 8.5 1区 化学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY
Wen Peng, Gen Wan, Sufang Cheng, Guyue Li, Liling Liu, Juan Chen, Pei Liu, Xiaoquan Guo, Xiaona Gao, Gaofeng Cai, Zhanhong Zheng, Ping Liu
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引用次数: 0

摘要

由于缺乏抗鸡肿瘤坏死因子受体相关因子2 (TRAF2)的特异性抗体,因此很难进一步探讨TRAF2在肺高压(PH)肉鸡肺动脉重构中的作用。本实验通过构建TRAF2重组蛋白原核表达载体制备了TRAF2的多克隆抗体,分析了TRAF2在肉仔鸡肺高压模型体内和体外的表达以及TRAF2对PASMCs活性和凋亡的影响。结果表明,用TRAF2重组蛋白免疫后获得高效价的多克隆抗体,黄芪多糖作为免疫佐剂可增强免疫效果。抗体特异性表明,该多克隆抗体与鸡、鸭的TRAF2蛋白特异性结合,而与家兔、小鼠和山羊的TRAF2蛋白弱结合。在肉仔鸡体内和体外PH模型中,TRAF2均显著上调。敲低TRAF2抑制PASMCs活性,诱导PASMCs凋亡。本研究为进一步研究肉鸡PH的发病机制奠定了基础,并为其防治和药物开发提供了新的靶点。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Potential role of TRAF2 in pulmonary hypertension in broiler chickens and preparation and specificity analysis of its polyclonal antibody.

Due to the lack of specific antibody anti-chicken tumor necrosis factor receptor-associated factor 2 (TRAF2), it is difficult to further explore the role of TRAF2 in pulmonary artery remodeling in pulmonary hypertension(PH) in broilers. In this experiment, we prepared a polyclonal antibody to TRAF2 by constructing a TRAF2 recombinant protein prokaryotic expression vector and analyzed the expression of TRAF2 in in vivo and in vitro models of pulmonary hypertension in broiler chickens and the effect of TRAF2 on the activity and apoptosis of PASMCs. The results showed that after immunization with TRAF2 recombinant protein we obtained high titers of polyclonal antibodies, and astragalus polysaccharide as an immune adjuvant could enhance the effect of immunization. Antibody specificity showed that the TRAF2 polyclonal antibody specifically bound to TRAF2 protein in chickens and ducks but weakly to TRAF2 protein in rabbits, mice and goats.TRAF2 was significantly upregulated in an in vivo and in vitro model of PH in broilers. Knockdown of TRAF2 inhibited the activity of PASMCs and induced apoptosis in PASMCs. Our study lays the foundation for further research on the pathomechanism of PH in broiler chickens and provides new targets for its prevention and drug development.

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来源期刊
International Journal of Biological Macromolecules
International Journal of Biological Macromolecules 生物-生化与分子生物学
CiteScore
13.70
自引率
9.80%
发文量
2728
审稿时长
64 days
期刊介绍: The International Journal of Biological Macromolecules is a well-established international journal dedicated to research on the chemical and biological aspects of natural macromolecules. Focusing on proteins, macromolecular carbohydrates, glycoproteins, proteoglycans, lignins, biological poly-acids, and nucleic acids, the journal presents the latest findings in molecular structure, properties, biological activities, interactions, modifications, and functional properties. Papers must offer new and novel insights, encompassing related model systems, structural conformational studies, theoretical developments, and analytical techniques. Each paper is required to primarily focus on at least one named biological macromolecule, reflected in the title, abstract, and text.
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