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QS-CATH 是一种从中国棘蛙(Quasipaa spinosa)中分离出来的有前途的治疗剂,评估其抗菌和免疫调节活性。

Assessment of the antimicrobial and immunomodulatory activity of QS-CATH, a promising therapeutic agent isolated from the Chinese spiny frogs (Quasipaa spinosa).

机构信息

College of Animal Sciences, Zhejiang University, Hangzhou 310058, China; Ecological Forestry Development Center of Suichang County, Lishui 323000, China.

College of Animal Sciences, Zhejiang University, Hangzhou 310058, China; Ecological Forestry Development Center of Suichang County, Lishui 323000, China.

出版信息

Comp Biochem Physiol C Toxicol Pharmacol. 2024 Sep;283:109943. doi: 10.1016/j.cbpc.2024.109943. Epub 2024 May 27.

Abstract

Cathelicidins are important antimicrobial peptides in various vertebrate species where they are crucial parts of the innate immune system. The current understanding of amphibian cathelicidins is limited, particularly with regard to their immunomodulatory effects. To address this knowledge gap, we produced the cDNA sequence of the cathelicidin gene from a skin transcriptome of the Chinese spiny frog Quasipaa spinosa. The amino acid sequence of the Quasipaa spinosa cathelicidin (QS-CATH) was predicted to consist of a signal peptide, a cathelin domain, and a mature peptide. Comparative analysis of the QS-CATH amino acid sequence with that of other amphibian cathelicidins revealed high variability in the functional mature peptide among amphibians, whereas the cathelin domain was conserved. The QS-CATH gene was expressed in several tissues, with the highest level of expression in the spleen. Upregulation of QS-CATH after Aeromonas hydrophila infection occurred in the kidney, gut, spleen, skin, and liver. Chemically synthesized QS-CATH exhibited pronounced antibacterial activity against Shigella flexneri, Staphylococcus warneri, Escherichia coli, Salmonella enterica, and Listeria monocytogenes. Furthermore, QS-CATH disrupted the cell membrane integrity of S. flexneri, as evidenced by a lactate dehydrogenase release assay, and it hydrolyzed the genomic DNA of S. flexneri. Additionally, QS-CATH elicited chemotaxis and modulated the expression of inflammatory cytokine genes in RAW264.7 mouse leukemic monocyte/macrophage cells. These findings confirm the antimicrobial effects of amphibian cathelicidin and its ability to influence immune cell function. This will expedite the potential utilization of amphibian antimicrobial peptides as therapeutic agents.

摘要

抗菌肽是各种脊椎动物物种中重要的抗菌肽,是先天免疫系统的重要组成部分。目前对两栖动物抗菌肽的了解有限,特别是它们的免疫调节作用。为了弥补这一知识空白,我们从中国棘蛙(Quasipaa spinosa)的皮肤转录组中获得了抗菌肽基因的 cDNA 序列。预测棘蛙抗菌肽(QS-CATH)的氨基酸序列由信号肽、抗菌肽前体和成熟肽组成。与其他两栖动物抗菌肽的氨基酸序列比较分析表明,两栖动物抗菌肽的成熟肽在功能上高度多样化,而抗菌肽前体则保守。QS-CATH 基因在多种组织中表达,在脾脏中的表达水平最高。在感染嗜水气单胞菌后,QS-CATH 在肾脏、肠道、脾脏、皮肤和肝脏中的表达上调。化学合成的 QS-CATH 对福氏志贺菌、华纳氏葡萄球菌、大肠杆菌、肠炎沙门氏菌和单核细胞增生李斯特菌表现出明显的抗菌活性。此外,QS-CATH 通过乳酸脱氢酶释放试验破坏了福氏志贺菌的细胞膜完整性,并水解了福氏志贺菌的基因组 DNA。此外,QS-CATH 还能引起 RAW264.7 小鼠白血病单核/巨噬细胞细胞的趋化作用,并调节炎症细胞因子基因的表达。这些发现证实了两栖动物抗菌肽的抗菌作用及其影响免疫细胞功能的能力。这将加速两栖动物抗菌肽作为治疗剂的潜在应用。

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