目的:本观察性研究探讨Pim-1在炎症信号通路中的调控机制。
方法:使用THP-1、RAW264.7、BV2和Jurkat人T细胞系。
方法:无。
方法:用脂多糖(LPS)诱导炎症,其次是PIM1敲除。蛋白质印迹,免疫沉淀,免疫荧光,和RT-PCR测定用于评估PIM1敲低对LPS诱导的炎症的影响。
结果:巨噬细胞样THP-1细胞中PIM1敲低抑制LPS诱导的促炎细胞因子的上调,诱导型一氧化氮合酶,环氧合酶-2,磷酸化Janus激酶,信号转导和转录激活因子3,细胞外信号调节激酶,c-Jun氨基末端激酶,p38和核因子κBp65(NF-κBp65)。它还抑制了NF-κB激酶α/β抑制剂的上调,并增强了NF-κBp65的核易位。此外,它抑制了NOD样受体家族pyrin结构域3(NLRP3)的上调,并抑制了LPS与三磷酸腺苷共同处理诱导的caspase-1的裂解。此外,p-转化生长因子-β激活激酶1(TAK1)与Pim-1相互作用。Pim激酶的所有三个成员(Pim-1,Pim-2和Pim-3)都是LPS介导的巨噬细胞炎症所必需的;然而,与Pim-1和Pim-3不同,Pim-2充当T细胞活性的负调节因子。
结论:Pim-1在LPS诱导的炎症反应中与TAK1相互作用,并参与MAPK/NF-κB/NLRP3信号通路。此外,考虑到Pim-2在T细胞中的负调节作用,需要进一步深入研究它们各自的功能。
OBJECTIVE: This observational study investigated the regulatory mechanism of Pim-1 in inflammatory signaling pathways.
METHODS: THP-1, RAW 264.7, BV2, and Jurkat human T cell lines were used.
METHODS: None.
METHODS: Lipopolysaccharide (LPS) was used to induce inflammation, followed by PIM1 knockdown. Western blot, immunoprecipitation, immunofluorescence, and RT-PCR assays were used to assess the effect of PIM1 knockdown on LPS-induced inflammation.
RESULTS: PIM1 knockdown in macrophage-like THP-1 cells suppressed LPS-induced upregulation of pro-inflammatory cytokines, inducible nitric oxide synthase, cyclooxygenase-2, phosphorylated Janus kinase, signal transducer and activator of transcription 3, extracellular signal-regulated kinase, c-Jun N-terminal kinase, p38, and nuclear factor kappa B p65 (NF-κB p65). It also suppressed upregulation of inhibitor of NF-κB kinase α/β and enhanced the nuclear translocation of NF-κB p65. Moreover, it inhibited the upregulation of Nod-like receptor family pyrin domain-containing 3 (NLRP3) and cleavage of caspase-1 induced by co-treatment of LPS with adenosine triphosphate. Additionally, p-transforming growth factor-β-activated kinase 1 (TAK1) interacted with Pim-1. All three members of Pim kinases (Pim-1, Pim-2, and Pim-3) were required for LPS-mediated inflammation in macrophages; however, unlike Pim-1 and Pim-3, Pim-2 functioned as a negative regulator of T cell activity.
CONCLUSIONS: Pim-1 interacts with TAK1 in LPS-induced inflammatory responses and is involved in MAPK/NF-κB/NLRP3 signaling pathways. Additionally, considering the negative regulatory role of Pim-2 in T cells, further in-depth studies on their respective functions are needed.