Tumor necrosis factor-alpha mediates pancreatitis responses in acinar cells via protein kinase C and proline-rich tyrosine kinase 2.
Tumor necrosis factor-alpha mediates pancreatitis responses in acinar cells via protein kinase C and proline-rich tyrosine kinase 2.
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肿瘤坏死因子-α 通过蛋白激酶 C 和富含脯氨酸的酪氨酸激酶 2 介导腺泡细胞中的胰腺炎反应。
DOI：
10.1016/j.gastro.2005.05.005
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发表时间：
2005
期刊：
影响因子：
29.4
通讯作者：
Pandol,StephenJ
中科院分区：
文献类型：
--
作者：
Satoh,Akihiko;Gukovskaya,AnnaS;Edderkaoui,Mouad;Daghighian,MelissaS;ReeveJr,JosephR;Shimosegawa,Tooru;Pandol,StephenJ
背景和目的尽管肿瘤坏死因子 α 被认为是急性胰腺炎炎症反应的重要介质，但其在该疾病其他病理特征中的作用仍不清楚。我们研究了肿瘤坏死因子α在细胞骨架反应中的作用以及胰腺腺泡细胞中潜在的信号传导机制。方法在分离的大鼠胰腺腺泡和AR42J细胞中，我们通过罗丹明-鬼笔环肽测定了肿瘤坏死因子α对肌动蛋白细胞骨架的影响。使用药理学和分子方法，我们评估了蛋白激酶 C、Src 激酶和富含脯氨酸的酪氨酸激酶 2 在此过程中的参与情况。我们还研究了这些信号通路在肿瘤坏死因子-α诱导的核因子-κB激活和细胞凋亡中的作用。结果肿瘤坏死因子-α增加了腺泡细胞中富含脯氨酸的酪氨酸激酶2的酪氨酸磷酸化。广谱蛋白激酶 C 抑制剂和 Src 激酶抑制剂均抑制肿瘤坏死因子-α 诱导的富含脯氨酸的酪氨酸激酶 2 磷酸化，但酪氨酸残基不同。使用蛋白激酶 C 同工型特异性抑制剂和反义方法，我们发现蛋白激酶 C δ 和 ϵ 介导富含脯氨酸的酪氨酸激酶 2 酪氨酸磷酸化。肿瘤坏死因子-α 通过依赖蛋白激酶 C、Src 激酶和富含脯氨酸的酪氨酸激酶 2 的机制导致肌动蛋白细胞骨架解体。抑制蛋白激酶 C（而非 Src 激酶）可减少肿瘤坏死因子-α 诱导的细胞凋亡。此外，通过反义转染，我们发现蛋白激酶Cδ和ε，而不是富含脯氨酸的酪氨酸激酶2，介导肿瘤坏死因子α诱导的核因子-κB激活。结论肿瘤坏死因子-α激活富含脯氨酸的酪氨酸激酶2，引起细胞骨架紊乱和核因子-κB引起炎症反应，并且它 通过蛋白激酶 C 介导的不同机制触发细胞死亡信号传导。这些结果提供了对胰腺腺泡细胞中将肿瘤坏死因子 α 与急性胰腺炎关键过程联系起来的机制的见解。
Background & AimsAlthough tumor necrosis factor α is implicated as an important mediator of the inflammatory response in acute pancreatitis, its role in other pathologic features of the disease remains unknown. We investigated the role for tumor necrosis factor α in cytoskeletal responses and the underlying signaling mechanisms in pancreatic acinar cells.MethodsIn isolated rat pancreatic acini and AR42J cells, we determined the effect of tumor necrosis factor α on the actin cytoskeleton by rhodamine-phalloidin. Using pharmacological and molecular approaches, we assessed the involvement of protein kinase C, Src kinases, and proline-rich tyrosine kinase 2 in the process. We also studied the involvement of these signaling pathways in tumor necrosis factor-α-induced nuclear factor-κB activation and apoptosis.ResultsTumor necrosis factor-α increased the tyrosine phosphorylation of proline-rich tyrosine kinase 2 in acinar cells. The broad-spectrum protein kinase C inhibitor and the Src kinase inhibitor both inhibited tumor necrosis factor-α-induced proline-rich tyrosine kinase 2 phosphorylation, but at different tyrosine residues. Using protein kinase C isoform-specific inhibitors and the antisense approach, we showed that protein kinase C δ and ϵ mediate proline-rich tyrosine kinase 2 tyrosine phosphorylation. Tumor necrosis factor-α caused disorganization of the actin cytoskeleton by a mechanism dependent on protein kinase C, Src kinases, and proline-rich tyrosine kinase 2. Inhibition of protein kinase C, but not Src kinases, decreased tumor necrosis factor-α-induced apoptosis. Furthermore, with antisense transfections, we showed that protein kinase C δ and ϵ, but not proline-rich tyrosine kinase 2, mediate tumor necrosis factor α-induced nuclear factor-κB activation.ConclusionsTumor necrosis factor-α activates proline-rich tyrosine kinase 2 to cause cytoskeletal disorganization and nuclear factor-κB to cause inflammatory response, and it triggers cell death signaling through divergent mechanisms mediated by protein kinase C. The results provide insights into the mechanisms in pancreatic acinar cells that link tumor necrosis factor α to critical processes in acute pancreatitis.