IL-32通过激活FAT4介导的河马/YAP信号传导来加剧人核细胞中的代谢紊乱

炎症微环境中的细胞外基质（ECM）代谢疾病在椎间盘退化（IDD）的发病机理中起关键作用。据报道，白介素-32（IL-32）参与了各种炎性疾病的进展。但是，尚不清楚它是否参与核核（NP）细胞的基质代谢。因此，这项研究旨在研究IL-32在调节炎症微环境中ECM代谢方面的机制。 RNA-Seq用于鉴定炎症微环境中NP细胞中异常表达的基因。进行了蛋白质印迹，实时定量PCR，免疫组织化学和免疫荧光分析，以测量在人NP组织或未经肿瘤坏死因子-α（TNF-α）处理的人NP组织或NP细胞中IL-32和代谢标记的表达。在体内，将过表达IL-32的腺相关病毒注入大鼠的尾椎间盘中，以评估其对IDD的影响。通过免疫沉淀和质谱法鉴定出与IL-32相互作用的蛋白质。过表达IL-32或击倒脂肪非典型钙粘蛋白4（FAT4）的慢病毒，是使用YES相关蛋白（YAP）抑制剂 - 垂直蛋白（VP）来治疗人NP细胞，以探索IL-32的发病机理。在人NP组织中验证了河马/YAP信号传导活性。如临床样品中所示，在退化性NP组织中，IL-32表达显着上调。此外，在TNF-α诱导的退化性NP细胞中，IL-32明显过表达。 IL-32过表达诱导大鼠模型中的IDD进程。从机械上讲，炎症微环境中IL-32的升高增强了其与FAT4和哺乳动物无菌的相互作用，并促使MST1/2磷酸化促使MST1/2磷酸化，并激活Hippo/Yap信号通路，并引起Hippo/YAP信号通路，从而导致基质型细胞nplix nplix nplix nplix nplix nplix nplix nplix nplix nplix。我们的结果表明，IL-32通过FAT4/MST/YAP轴介导NP细胞中NP细胞中基质代谢疾病，为精确治疗IDD提供了理论基础。

Extracellular matrix (ECM) metabolism disorders in the inflammatory microenvironment play a key role in the pathogenesis of intervertebral disc degeneration (IDD). Interleukin-32 (IL-32) has been reported to be involved in the progression of various inflammatory diseases; however, it remains unclear whether it participates in the matrix metabolism of nucleus pulposus (NP) cells. Therefore, this study aimed to investigate the mechanism of IL-32 on regulating the ECM metabolism in the inflammatory microenvironment. RNA-seq was used to identify aberrantly expressed genes in NP cells in the inflammatory microenvironment. Western blotting, real-time quantitative PCR, immunohistochemistry and immunofluorescence analysis were performed to measure the expression of IL-32 and metabolic markers in human NP tissues or NP cells treated with or without tumor necrosis factor-α (TNF-α). In vivo, an adeno-associated virus overexpressing IL-32 was injected into the caudal intervertebral discs of rats to assess its effect on IDD. Proteins interacting with IL-32 were identified via immunoprecipitation and mass spectrometry. Lentivirus overexpressing IL-32 or knocking down Fat atypical cadherin 4 (FAT4), yes-associated protein (YAP) inhibitor-Verteporfin (VP) were used to treat human NP cells, to explore the pathogenesis of IL-32. Hippo/YAP signaling activity was verified in human NP tissues. IL-32 expression was significantly upregulated in degenerative NP tissues, as indicated in the clinical samples. Furthermore, IL-32 was remarkably overexpressed in TNF-α-induced degenerative NP cells. IL-32 overexpression induced IDD progression in the rat model. Mechanistically, the elevation of IL-32 in the inflammatory microenvironment enhanced its interactions with FAT4 and mammalian sterile 20-like kinase1/2 (MST1/2) proteins, prompting MST1/2 phosphorylation, and activating the Hippo/YAP signaling pathway, causing matrix metabolism disorder in NP cells. Our results suggest that IL-32 mediates matrix metabolism disorders in NP cells in the inflammatory micro-environment via the FAT4/MST/YAP axis, providing a theoretical basis for the precise treatment of IDD.