Neutrophil Chemotaxis and NETosis in Murine Chronic Liver Injury via Cannabinoid Receptor 1/Gαi/o/ROS/p38 MAPK Signaling Pathway
Neutrophil Chemotaxis and NETosis in Murine Chronic Liver Injury via Cannabinoid Receptor 1/Gαi/o/ROS/p38 MAPK Signaling Pathway
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小鼠慢性肝损伤中的中性粒细胞趋化性和 NETosis 通过大麻素受体 1/ Gi/o/ ROS/ p38 MAPK 信号通路
DOI：
10.3390/cells9020373
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发表时间：
2020-02-01
期刊：
影响因子：
6
通讯作者：
Li, Liying
中科院分区：
文献类型：
--
作者：
Zhou, Xuan;Yang, Le;Li, Liying
中性粒细胞在炎症性疾病的控制中发挥重要作用。然而，大麻素受体（CB）是否在中性粒细胞趋化性和NETosis在无菌性肝脏炎症中发挥作用仍然未知。通过FACS、免疫荧光、qRT-PCR和Western blot表征中性粒细胞上标志物基因的表达。中性粒细胞的数量显着升高，从7天，并在2周达到高峰，在四氯化碳（CCl 4）处理的小鼠肝脏。损伤肝组织中中性粒细胞标志物Ly 6 G的mRNA表达与CB 1、CB 2的表达呈正相关。CBs在体外培养的中性粒细胞中大量表达，CB 1激动剂ACEA促进了中性粒细胞的趋化性和细胞骨架重塑，而CB 1拮抗剂AM 281可抑制ACEA的趋化性和细胞骨架重塑。此外，ACEA诱导NETosis，从溶酶体释放髓过氧化物酶和ROS爆发，表明中性粒细胞活化，通过G α（i/o）。相反，CB 2激动剂JWH 133对中性粒细胞功能没有影响。ROS和p38 MAPK信号通路参与CB 1介导的中性粒细胞功能，ROS位于p38 MAPK的上游。CB 1阻断在体内显着减弱中性粒细胞浸润和肝脏炎症CCl 4处理的小鼠。综上所述，CB 1通过G α（i/o）/ROS/p38 MAPK信号通路介导肝脏炎症中的中性粒细胞趋化和NETosis，这代表了肝脏疾病的有效治疗策略。
Neutrophils play an essential role in the control of inflammatory diseases. However, whether cannabinoid receptors (CBs) play a role in neutrophil chemotaxis and NETosis in sterile liver inflammation remains unknown. The expression of marker genes on neutrophils was characterized by FACS, immunofluorescence, qRT-PCR, and Western blot. The amount of neutrophils was significantly elevated from 7 days and reached the peak at 2 weeks in carbon tetrachloride (CCl4)-treated mouse liver. The mRNA expression of neutrophil marker Ly6G had positive correlation with CB1 and CB2 expression in injured liver. In vitro CBs were abundantly expressed in isolated neutrophils and CB1 agonist ACEA promoted the chemotaxis and cytoskeletal remodeling, which can be suppressed by CB1 antagonist AM281. Moreover, ACEA induced NETosis, myeloperoxidase release from lysosome and ROS burst, indicating neutrophil activation, via G alpha(i/o). Conversely, CB2 agonist JWH133 had no effect on neutrophil function. ROS and p38 MAPK signaling pathways were involved in CB1-mediated neutrophil function, and ROS was upstream of p38 MAPK. CB1 blockade in vivo significantly attenuated neutrophil infiltration and liver inflammation in CCl4-treated mice. Taken together, CB1 mediates neutrophil chemotaxis and NETosis via G alpha(i/o)/ROS/p38 MAPK signaling pathway in liver inflammation, which represents an effective therapeutic strategy for liver diseases.