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ASC 在抗感染过程中的作用,作为黑鲷(Sebastes schlegelii)炎症小体的关键组成部分。

Role of ASC, a key component of the inflammasome in the antimicrobial process in black rockfish (Sebastes schlegelii).

机构信息

Key Laboratory of Marine Genetics and Breeding, Ministry of Education, Ocean University of China, 266003, Qingdao, Shandong, China.

Key Laboratory of Marine Genetics and Breeding, Ministry of Education, Ocean University of China, 266003, Qingdao, Shandong, China.

出版信息

Fish Shellfish Immunol. 2024 Nov;154:109886. doi: 10.1016/j.fsi.2024.109886. Epub 2024 Sep 6.

DOI:10.1016/j.fsi.2024.109886
PMID:39245187
Abstract

Apoptosis-associated speck-like protein containing a CARD (ASC) serves as a pivotal component within the inflammasome complex, playing a critical role in the activation of the innate immune response against pathogenic infection. However, the functional significance of inflammasome ASC in teleosts remains unclear. In this study, the coding sequence (CDS) region of ASC gene of Sebastes schlegelii (SsASC) was cloned, and we observed a high conservation of SsASC with teleosts through comprehensive bioinformatics analysis. SsASC and SsCaspase-1 were found to be highly expressed in immune tissues such as spleen and head kidney. Furthermore, our findings revealed that SsASC interacts with SsCaspase-1 through CARD-CARD interactions to generate oligomeric speck-like structures, whereas the PYD structural domain of SsASC forms only filamentous structures. To further understand the role of SsASC in combating Edwardsiella piscicida (E. piscicida) infection, we developed a SsASC knockdown model using in vivo siRNA injection and E. piscicida challenge via intraperitoneal injection. The model demonstrated that E. piscicida infection up-regulated SsASC expression, which was markedly reduced upon SsASC knockdown. Concurrently, E. piscicida colonization was significantly enhanced in the knockdown group, accompanied by a suppression of inflammatory factor expression. These findings confirm the pivotal antibacterial and anti-infective role of SsASC in the Sebastes schlegelii immune response upon E. piscicida stimulation. Our study highlights the significance of SsASC in the innate immune defense mechanism of teleosts against bacterial pathogens.

摘要

凋亡相关斑点样蛋白包含一个 CARD(ASC),作为炎症小体复合物中的关键组成部分,在针对病原感染的固有免疫反应的激活中发挥着关键作用。然而,炎症小体 ASC 在硬骨鱼类中的功能意义尚不清楚。在本研究中,克隆了Sebastes schlegelii(SsASC)的 ASC 基因的编码序列(CDS)区,通过全面的生物信息学分析,我们观察到 SsASC 与硬骨鱼类具有高度的保守性。发现 SsASC 和 SsCaspase-1 在免疫组织如脾脏和头肾中高度表达。此外,我们的研究结果表明,SsASC 通过 CARD-CARD 相互作用与 SsCaspase-1 相互作用,产生寡聚斑点样结构,而 SsASC 的 PYD 结构域仅形成丝状结构。为了进一步了解 SsASC 在抵御爱德华氏菌(E. piscicida)感染中的作用,我们使用体内 siRNA 注射和通过腹腔注射 E. piscicida 挑战开发了 SsASC 敲低模型。该模型表明,E. piscicida 感染上调了 SsASC 的表达,而 SsASC 敲低后其表达显著降低。同时,在敲低组中,E. piscicida 定植显著增强,伴随着炎症因子表达的抑制。这些发现证实了 SsASC 在 Sebastes schlegelii 对 E. piscicida 刺激的免疫反应中作为关键抗菌和抗感染作用的重要性。我们的研究强调了 SsASC 在硬骨鱼类针对细菌病原体的固有免疫防御机制中的重要性。

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