National Engineering Research Center of Marine Facilities Aquaculture, Marine Science and Technology College, Zhejiang Ocean University, Zhoushan 316004, China.
Int J Mol Sci. 2023 Mar 21;24(6):5928. doi: 10.3390/ijms24065928.
Interleukin-17 (IL-17) represents a class of proinflammatory cytokines involved in chronic inflammatory and degenerative disorders. Prior to this study, it was predicted that an IL-17 homolog could be targeted by -novel_miR_145 to participate in the immune response of . This study employed a variety of molecular and cell biology research methods to explore the association between -novel_miR_145 and IL-17 homolog and their immunomodulatory effects. The bioinformatics prediction confirmed the affiliation of the IL-17 homolog with the mussel IL-17 family, followed by quantitative real-time PCR assays (qPCR) to demonstrate that IL-17-3 was highly expressed in immune-associated tissues and responded to bacterial challenges. Results from luciferase reporter assays confirmed the potential of IL-17-3 to activate downstream NF-κb and its targeting by -novel_miR_145 in HEK293 cells. The study also produced IL-17-3 antiserum and found that -novel_miR_145 negatively regulates IL-17-3 via western blotting and qPCR assays. Furthermore, flow cytometry analysis indicated that -novel_miR_145 negatively regulated IL-17-3 to alleviate LPS-induced apoptosis. Collectively, the current results showed that IL-17-3 played an important role in molluscan immune defense against bacterial attack. Furthermore, IL-17-3 was negatively regulated by -novel_miR_145 to participate in LPS-induced apoptosis. Our findings provide new insights into noncoding RNA regulation in invertebrate models.
白细胞介素-17(IL-17)代表了一类参与慢性炎症和退行性疾病的促炎细胞因子。在这项研究之前,人们预测一种 IL-17 同源物可以被新型 miR-145 靶向,以参与的免疫反应。本研究采用多种分子和细胞生物学研究方法,探讨新型 miR-145 与 IL-17 同源物及其免疫调节作用之间的关系。生物信息学预测证实了 IL-17 同源物与贻贝 IL-17 家族的关联,随后进行定量实时 PCR 分析(qPCR)以证明 IL-17-3 在免疫相关组织中高度表达,并对细菌挑战做出反应。荧光素酶报告基因检测结果证实了 IL-17-3 激活下游 NF-κb 的潜力及其在 HEK293 细胞中被新型 miR-145 靶向的潜力。该研究还产生了 IL-17-3 抗血清,并通过 Western blot 和 qPCR 分析发现新型 miR-145 通过负向调节 IL-17-3。此外,流式细胞术分析表明新型 miR-145 通过负向调节 IL-17-3 来减轻 LPS 诱导的细胞凋亡。总之,目前的结果表明 IL-17-3 在软体动物免疫防御细菌攻击中发挥重要作用。此外,IL-17-3 受到新型 miR-145 的负向调节,以参与 LPS 诱导的细胞凋亡。我们的研究结果为无脊椎动物模型中的非编码 RNA 调控提供了新的见解。