Cannabinoid Receptor 1 Participates in Liver Inflammation by Promoting M1 Macrophage Polarization via RhoA/NF-κB p65 and ERK1/2 Pathways, Respectively, in Mouse Liver Fibrogenesis.
Cannabinoid Receptor 1 Participates in Liver Inflammation by Promoting M1 Macrophage Polarization via RhoA/NF-κB p65 and ERK1/2 Pathways, Respectively, in Mouse Liver Fibrogenesis.
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在小鼠肝脏纤维形成中，大麻素受体 1 分别通过 RhoA/NF-kappa B p65 和 ERK1/2 途径促进 M1 巨噬细胞极化，从而参与肝脏炎症
DOI：
10.3389/fimmu.2017.01214
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发表时间：
2017
影响因子：
7.3
通讯作者：
Li L
中科院分区：
文献类型：
--
作者：
Tian L;Li W;Yang L;Chang N;Fan X;Ji X;Xie J;Yang L;Li L
巨噬细胞M1/M2极化介导组织损伤和炎症反应。大麻素受体(CB)1通过影响骨髓来源的单核/巨噬细胞(BMMS)激活参与肝纤维化的形成。然而，关于CB1是否参与BMMS的极化的了解仍然有限。我们发现，在四氯化碳(CCl_4)诱导的小鼠肝脏损伤中，M1基因特征(包括CD86、MIP-1β、肿瘤坏死因子、IL-6和诱导型一氧化氮合酶)和M1巨噬细胞(CD86+细胞，经F4/80门控)的数量显著增加，而通过RT-qPCR和荧光激活细胞分类检测，M2型巨噬细胞的数量几乎没有变化。我们先前的研究证实CB1参与了CCl4诱导的肝纤维化。结果表明，CB1的表达与CD86呈正相关。用其拮抗剂或siRNA在体内阻断CB1，通过RT-qPCR、Western印迹和细胞珠阵(CBA)检测，下调M1标志物的mRNA和蛋白水平，并减少M1巨噬细胞的比例。此外，通过接受绿色荧光蛋白转基因小鼠骨髓移植的嵌合体小鼠模型和库普弗细胞耗尽的氯膦酸盐脂质体注射模型，研究了CB1在库普弗细胞和骨髓基质细胞极化中的作用。我们发现CB1特别参与了BMM向M1表型的极化，但对Kupffer细胞的极化没有影响。其原因可能是Kupffer细胞中CB1的表达低于BMMS。在体外，我们发现CB1参与了BMMS对M1的极化。Ptx[G(α)I/o蛋白抑制剂]、Y27632(ROCK抑制剂)和PD98059[细胞外信号调节激酶(ERK)抑制剂]可明显减弱CB1诱导的M1极化，而p38抑制剂SB203580和化合物C(AMPK抑制剂)则无此作用。ACEA(CB1激动剂)激活G(α)I/O偶联CB1，然后独立地扩大GTP结合的Rho和磷酸化ERK1/2。NF-κB p65核转位也是M1表型巨噬细胞的标志。我们发现，Cb1仅通过G(κ)I/o/RhoA信号通路启动了NF-αB p65核转位。CB1通过G(α)I/o/RhoA/NF-κBp65和G(α)I/o/ERK1/2两条独立的信号通路在调节肝损伤时骨髓基质细胞M1极化中发挥重要作用。
Macrophage M1/M2 polarization mediates tissue damage and inflammatory responses. Cannabinoid receptor (CB) 1 participated in liver fibrogenesis by affecting bone marrow (BM)-derived monocytes/macrophages (BMMs) activation. However, the knowledge of whether CB1 is involved in the polarization of BMMs remains limited. Here, we found M1 gene signatures (including CD86, MIP-1β, tumor necrosis factor, IL-6, and inducible nitric oxide synthase) and the amount of M1 macrophages (CD86+ cells, gated by F4/80) were significantly elevated in carbon tetrachloride (CCl4)-induced mouse injured livers, while that of M2 type macrophages had little change by RT-qPCR and fluorescence-activated cell sorting (FACS). Our preceding study confirmed CB1 was involved in CCl4-induced liver fibrogenesis. Our results noted CB1 expression showed positive correlation with CD86. Blockade of CB1 by its antagonist or siRNA in vivo downregulated the mRNA and protein levels of M1 markers using RT-qPCR, western blot, and Cytometric Bead Array (CBA) assays, and reduced the proportion of M1 macrophages. Moreover, chimera mouse models, which received BM transplants from EGFP-transgenic mice or clodronate liposome injection mouse models, in which Kupffer cells were depleted, were performed to clarify the role of CB1 on the polarization of Kupffer cells and BMMs. We found that CB1 was especially involved in BMM polarization toward M1 phenotype but have no effect on that of Kupffer cells. The reason might due to the lower CB1 expression in Kupffer cells than that of BMMs. In vitro, we discovered CB1 was involved in the polarization of BMMs toward M1. Furthermore, CB1-induced M1 polarization was apparently impaired by PTX [G(α)i/o protein inhibitor], Y27632 (ROCK inhibitor), and PD98059 [extracellular signal-regulated kinase (ERK) inhibitor], while SB203580 (p38 inhibitor) and compound C (AMPK inhibitor) had no such effect. ACEA (CB1 agonist) activated G(α)i/o coupled CB1, then enlarged GTP-bound Rho and phosphor-ERK1/2, independently. NF-κB p65 nuclear translocation is also a marker of M1 phenotype macrophages. We found that CB1 switched on NF-κB p65 nuclear translocation only depending on G(α)i/o/RhoA signaling pathway. CB1 plays a crucial role in regulating M1 polarization of BMMs in liver injury, depending on two independent signaling pathways: G(α)i/o/RhoA/NF-κB p65 and G(α)i/o/ERK1/2 pathways.
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影响因子：
4.8
作者：
Morell C;Bort A;Vara D;Ramos-Torres A;Rodríguez-Henche N;Díaz-Laviada I
通讯作者：
Díaz-Laviada I
影响因子：
13.5
作者：
Cinar, Resat;Godlewski, Grzegorz;Liu, Jie;Tam, Joseph;Jourdan, Tony;Mukhopadhyay, Bani;Harvey-White, Judith;Kunos, George
通讯作者：
Kunos, George
影响因子：
82.9
作者：
通讯作者：
--
影响因子：
19.6
作者：
Lecru, Lola;Desterke, Christophe;Francois, Helene
通讯作者：
Francois, Helene
影响因子：
4.8
作者：
Martella, Andrea;Silvestri, Cristoforo;Carnevali, Oliana
通讯作者：
Carnevali, Oliana