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制备抗 Toll 样受体 4 纳米抗体及其对革兰氏阴性脓毒症的作用。

Preparation of Anti Toll-Like Receptor-4 Nano-Antibody and Its Effect on Gram Negative Sepsis.

机构信息

Department of Intensive Care Unit, The 2nd Affiliated Hospital of Chengdu Medical College Nuclear Industry 416 Hospital, Chengdu, 610000, China.

Department of Emergency, The 2nd Affiliated Hospital of Chengdu Medical College Nuclear Industry 416 Hospital, Chengdu, 610000, China.

出版信息

J Nanosci Nanotechnol. 2021 Feb 1;21(2):1048-1053. doi: 10.1166/jnn.2021.18664.

Abstract

The essence of sepsis is the excessive immune response caused by infectious factors, which leads to the release of a large number of inflammatory factors and the injury of tissues and organs. Once the reaction is triggered, it will develop according to its own rules, and it does not depend on infection factors. The occurrence and severity of sepsis depend entirely on the reaction of the body. When pathogens invade the body, they rely on pattern recognition receptors to play a defensive role, TLR4 is the main switch of innate immunity. The current research shows that LPS can induce the up-regulation of TLR4 gene expression in monocytes, neutrophils and other immune cells. The upregulated TLR4, as a new receptor, mediates more cell activation and the release of inflammatory mediators. Under this positive feedback, the inflammatory response is constantly amplified, which eventually leads to the development of sepsis. In this study, TLR4 extracellular domain antigen was obtained by protein expression and purification, and anti-TLR4 C-terminal and intermediate domain nano-antibody was obtained by phage display antibody library preparation technology. and experiments confirmed that anti-TLR4 nano-antibody can effectively reduce the release of inflammatory factors and improve the survival rate of animals, while the C-terminal and intermediate domain are closed at the same time, the effect is more obvious. The clinical treatment of sepsis provides new ideas and strategies.

摘要

脓毒症的本质是感染因素引起的过度免疫反应,导致大量炎症因子的释放和组织器官的损伤。一旦反应被触发,它将按照自己的规则发展,而不依赖于感染因素。脓毒症的发生和严重程度完全取决于机体的反应。当病原体侵入人体时,它们依靠模式识别受体发挥防御作用,TLR4 是先天免疫的主要开关。目前的研究表明,LPS 可以诱导单核细胞、中性粒细胞等免疫细胞中 TLR4 基因表达的上调。上调的 TLR4 作为一种新的受体,介导更多的细胞激活和炎症介质的释放。在这种正反馈下,炎症反应不断放大,最终导致脓毒症的发生。在本研究中,通过蛋白质表达和纯化获得了 TLR4 胞外结构域抗原,通过噬菌体展示抗体库制备技术获得了抗 TLR4 C 端和中间结构域纳米抗体。并通过实验证实抗 TLR4 纳米抗体可有效减少炎症因子的释放,提高动物的存活率,而同时封闭 C 端和中间结构域时,效果更为明显。为脓毒症的临床治疗提供了新的思路和策略。

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