GRK2 Mediated Abnormal Transduction of PGE2-EP4-cAMP-CREB Signaling Induces the Imbalance of Macrophages Polarization in Collagen-Induced Arthritis Mice
GRK2 Mediated Abnormal Transduction of PGE2-EP4-cAMP-CREB Signaling Induces the Imbalance of Macrophages Polarization in Collagen-Induced Arthritis Mice
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GRK2 介导的 PGE2-EP4-cAMP-CREB 信号异常转导导致胶原诱导关节炎小鼠巨噬细胞极化失衡
DOI：
10.3390/cells8121596
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发表时间：
2019-12-01
期刊：
影响因子：
6
通讯作者：
Wei, Wei
中科院分区：
文献类型：
--
作者：
Yang, Xuezhi;Li, Susu;Wei, Wei
类风湿性关节炎（rheologicarthritis，RA）是以滑膜内大量慢性炎症细胞浸润为特征的疾病。RA患者关节液中巨噬细胞以M1为主，巨噬细胞极化失衡的机制尚未完全阐明。前列腺素E2（PGE 2）部分地通过环磷酸腺苷（cAMP）-环AMP反应元件结合（CREB）信号传导增强M2极化。然而，以往的研究发现，PGE 2持续刺激佐剂性关节炎大鼠成纤维样滑膜细胞可引起cAMP的减少，这主要是由G蛋白偶联受体激酶2（GRK 2）诱导的EP 4过度脱敏引起的。GRK 2介导的EP 4过度脱敏是否会降低RA中cAMP的水平并抑制M2极化尚不清楚。我们观察到M1巨噬细胞在胶原诱导的关节炎（CIA）小鼠的腹腔巨噬细胞（PMs）、骨髓源性巨噬细胞（BBM）和滑膜巨噬细胞中占优势。PGE 2可通过EP 4-cAMP-CREB促进正常小鼠PM的M2极化，但对CIA小鼠PM的M2极化无促进作用。此外，我们发现PGE 2刺激的EP 4过度脱敏导致PGE 2-cAMP-CREB信号异常以及巨噬细胞极化失衡。通过GRK 2 siRNA或CRISPR/Cas9靶向破坏Raw264.7（RAW）中的GRK 2下调M1巨噬细胞标志物，上调M2巨噬细胞标志物和EP 4膜定位。在GRK 2 +/−小鼠的BMPs和PM中观察到M1/M2比值降低和p-CREB表达增加。本研究揭示了GRK 2在调节RA巨噬细胞功能中的新作用，为RA的精确治疗提供了新的思路。
Rheumatoid arthritis (RA) is characterized by the massive infiltration of various chronic inflammatory cells in synovia. In synovial fluid of patients with RA, M1 macrophages are dominant among all subtypes of macrophages, the mechanisms of macrophages polarization imbalance in RA has not been fully illuminated. The prostaglandin E2 (PGE2) augments M2 polarization in part via the cyclic adenosine monophosphate (cAMP)-cyclic AMP responsive element binding (CREB) signaling. However, previous study found constant stimulus of PGE2 on fibroblast-like synovial cells of adjuvant arthritis rats induced the decrease of cAMP, which is primarily caused by G protein-coupled receptor kinase 2 (GRK2)-induced EP4 over- desensitization. Whether GRK2 mediated-EP4 over-desensitization reduces the level of cAMP and inhibits M2 polarization in RA is unclear. Here we observed M1 macrophages were dominant in peritoneal macrophages (PMs), bone-marrow-derived macrophages (BMMs) and synovial macrophages of collagen-induced arthritis (CIA) mice. PGE2 stimulated M2 polarization via the EP4-cAMP-CREB in normal mice, while failed to promote M2 polarization in the PMs of CIA mice. Further, we found the EP4 over-desensitization stimulated by PGE2 induced abnormal PGE2-cAMP-CREB signaling as well as the imbalance of macrophage polarization. Targeted disruption of GRK2 in Raw264.7 (RAW) through GRK2 siRNA or CRISPR/Cas9 downregulated the M1 macrophage markers, upregulated the M2 macrophage markers and the EP4 membrane localization. The reduced M1/M2 ratio and increased p-CREB expression were observed in BMMs and PMs of GRK2+/− mice. This study highlighted a novel role of GRK2 in regulating macrophages function in RA and provided new idea for precision treatment of RA.