Background: Formyl peptide receptor 2 (FPR2) is involved in the pathogenesis of chronic inflammatory diseases, being activated either by pro-resolving or proinflammatory ligands. FPR2-associated signal transduction pathways result in phosphorylation of several proteins and in NADPH oxidase activation. We, herein, investigated molecular mechanisms underlying phosphorylation of heat shock protein 27 (HSP27), oxidative stress responsive kinase 1 (OSR1), and myristolated alanine-rich C-kinase substrate (MARCKS) elicited by the pro-resolving FPR2 agonists WKYMVm and annexin A1 (ANXA1). Methods: CaLu-6 cells or p22phox^Crispr/Cas9 double nickase CaLu-6 cells were incubated for 5 min with WKYMVm or ANXA1, in the presence or absence of NADPH oxidase inhibitors. Phosphorylation at specific serine residues of HSP27, OSR1, and MARCKS, as well as the respective upstream kinases activated by FPR2 stimulation was analysed. Results: Blockade of NADPH oxidase functions prevents WKYMVm- and ANXA1-induced HSP-27(Ser82), OSR1(Ser339) and MARCKS(Ser170) phosphorylation. Moreover, NADPH oxidase inhibitors prevent WKYMVm- and ANXA1-dependent activation of p38MAPK, PI3K and PKCδ, the kinases upstream to HSP-27, OSR1 and MARCKS, respectively. The same results were obtained in p22phox^Crispr/Cas9 cells. Conclusions: FPR2 shows an immunomodulatory role by regulating proinflammatory and anti-inflammatory activities and NADPH oxidase is a key regulator of inflammatory pathways. The activation of NADPH oxidase-dependent pro-resolving downstream signals suggests that FPR2 signalling and NADPH oxidase could represent novel targets for inflammation therapeutic intervention.

中文翻译：

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亲解析的FPR2激动剂调节HSP27，OSR1和MARCKS的NADPH氧化酶依赖性磷酸化以及相应上游激酶的激活

背景：甲酰基肽受体2（FPR2）参与慢性炎症性疾病的发病机理，通过促分辨或促炎性配体激活。FPR2相关的信号转导途径导致几种蛋白质的磷酸化和NADPH氧化酶激活。我们在这里研究了由热分解FPR2激动剂WKYMVm和膜联蛋白引起的热休克蛋白27（HSP27），氧化应激反应激酶1（OSR1）和富含肉豆蔻酸的富含丙氨酸的C激酶底物（MARCKS）磷酸化的分子机制。 A1（ANXA1）。方法：CaLu-6细胞或p22phox ^{Crispr / Cas9}在存在或不存在NADPH氧化酶抑制剂的情况下，将双切口酶CaLu-6细胞与WKYMVm或ANXA1孵育5分钟。分析了HSP27，OSR1和MARCKS的特定丝氨酸残基的磷酸化，以及通过FPR2刺激激活的相应上游激酶。结果：NADPH氧化酶功能的阻止可防止WKYMVm-和ANXA1诱导的HSP-27（Ser82），OSR1（Ser339）和MARCKS（Ser170）磷酸化。此外，NADPH氧化酶抑制剂可分别阻止HSP-27，OSR1和MARCKS上游激酶p38MAPK，PI3K和PKCδ的WKYMVm依赖性和ANXA1依赖性活化。在p22phox ^{Crispr / Cas9}中获得了相同的结果细胞。结论：FPR2通过调节促炎和抗炎活性显示出免疫调节作用，而NADPH氧化酶是炎性途径的关键调节剂。NADPH氧化酶依赖性促分解下游信号的激活表明FPR2信号和NADPH氧化酶可能代表炎症治疗干预的新目标。