Suppression of macrophage migration by down-regulating Src/FAK/P130Cas activation contributed to the anti-inflammatory activity of sinomenine
Suppression of macrophage migration by down-regulating Src/FAK/P130Cas activation contributed to the anti-inflammatory activity of sinomenine
复制标题
通过下调 Src/FAK/P130Cas 激活抑制巨噬细胞迁移有助于青藤碱的抗炎活性
DOI：
10.1016/j.phrs.2021.105513
复制
发表时间：
2021-04-23
影响因子：
9.3
通讯作者：
Zhou, Hua
中科院分区：
文献类型：
--
作者：
Gao, Wan-Jiao;Liu, Jian-Xin;Zhou, Hua
炎症部位的大量巨噬细胞不仅放大了炎症反应的严重程度，而且还有助于许多慢性炎症性疾病、自身免疫性疾病和癌症的有害进展。巨噬细胞迁移是巨噬细胞进入炎症部位和参与病理过程的先决条件。因此，抑制巨噬细胞迁移是一种潜在的抗炎机制。此外，炎症的缓解也可以阻止巨噬细胞的浸润。青藤碱（Sinomenine）是一种从中国药用植物青藤中提取的生物碱。它具有多种药理作用，包括抗炎症、免疫抑制和抗关节炎。然而，其抗炎分子机制和对巨噬细胞迁移的影响尚不完全清楚。本研究旨在探讨SIN对巨噬细胞体内外迁移的药理作用和分子机制，并阐明其与巨噬细胞迁移相关的抗炎机制。我们的研究结果表明，在体外，SIN减少了RAW264.7细胞向炎症爪迁移的数量，并阻断了脂多糖（LPS）诱导的RAW264.7细胞和骨髓源性巨噬细胞（bmdm）的迁移。此外，SIN还能减弱BMDMs的三维间质迁移。循环和外周巨噬细胞耗竭后巨噬细胞迁移的缺失导致炎症反应严重程度的降低。在巨噬细胞缺失（巨噬细胞（-/-））小鼠中，随着炎症严重程度的降低，RAW264.7细胞的迁移受到抑制。在巨噬细胞-/-小鼠中，SIN对炎症反应的影响不明显，但仍观察到SIN对RAW264.7细胞迁移的抑制作用。此外，在正常小鼠中，经SIN预处理的RAW264.7细胞的迁移受到抑制。最后，支持巨噬细胞间质迁移的Src/focal adhesion kinase (FAK)/P130Cas轴激活以及促进Src/FAK/P130Cas激活的iNOS表达、NO生成、整合素α V和整合素β 3表达均被SIN下调。然而，SIN对单核细胞趋化蛋白-1 （MCP-1）的表达没有明显影响，MCP-1是巨噬细胞迁移的重要趋化因子。这些结果表明，SIN通过下调Src/FAK/P130Cas轴的激活，显著抑制巨噬细胞间质迁移。炎症反应与巨噬细胞迁移之间存在相互调节的相关性，而SIN对巨噬细胞迁移的影响可能与其抗炎活性有关。
A large number of macrophages in inflamed sites not only amplify the severity of inflammatory responses but also contribute to the deleterious progression of many chronic inflammatory diseases, autoimmune diseases and cancers. Macrophage migration is a prerequisite for their entry into inflammatory sites and their participation of macrophages in the pathologic processes. Inhibition of macrophage migration is therefore a potential anti-inflammatory mechanism. Moreover, alleviation of inflammation also prevents the macrophages infiltration. Sinomenine (SIN) is an alkaloid derived from the Chinese medicinal plant Sinomenium acutum. It has multiple pharmacological effects, including anti-inflammation, immunosuppression, and anti-arthritis. However, its anti-inflammatory molecular mechanisms and effect on macrophage migration are not fully understood. The purpose of this research was to investigate the pharmacological effects and the molecular mechanism of SIN on macrophage migration in vivo and in vitro as well as to elucidate its anti-inflammatory mechanisms associated with macrophage migration. Our results showed that SIN reduced the number of RAW264.7 cells migrating into inflammatory paws and blocked lipopolysaccharide (LPS)-induced RAW264.7 cells and bone marrow-derived macrophages (BMDMs) migration in vitro. Furthermore, SIN attenuated the 3D mesenchymal migration of BMDMs. The absence of macrophage migration after circulatory and periphery macrophages depletion led to a reduction in the severity of inflammatory response. In macrophages depleted (macrophages(-/-)) mice, as inflammatory severity decreased, RAW264.7 cells migration was suppressed. A non-obvious effect of SIN on the inflammatory response was found in macrophages-/- mice, while the inhibitory effect of SIN on RAW264.7 cells migration was still observed. Furthermore, the migration of RAW264.7 cells pre-treated with SIN was suppressed in normal mice. Finally, Src/focal adhesion kinase (FAK)/P130Cas axis activation, which supports macrophages mesenchymal migration, and iNOS expression, NO production, integrin alpha V and in integrin beta 3 expressions, which promote Src/FAK/P130Cas activation, were down-regulated by SIN. However, SIN had no obvious effect on the expression of the monocyte chemoattractant protein-1 (MCP-1), which is an important chemokine for macrophage migration. These results indicated that SIN significantly inhibited macrophage mesenchymal migration by down-regulating on Src/FAK/P130Cas axis activation. There was a mutual regulatory correlation between the inflammatory response and macrophage migration, and the effects of SIN on macrophage migration were involved in its anti-inflammatory activity.