大麻素受体1通过PI3K/AKT信号通路介导单核巨噬细胞向M1型极化  

Study of Cannabinoid Receptor 1 Mediated Monocytes/Macrophages M1 Polarization via PI3K/AKT Signaling Pathway

作　　者：杨琳[1,2] 田蕾 常娜[1,2] 段向辉 李丽英 Yang Lin;Tian Lei;Chang Na;Duan Xianghui;Li Liying(Departrnent of Cell Biology,Capital Medical University,Beijing 100069,China;Municipal Laboratory for 'Liver Protection and Regulation of Regeneration',Beijing 100069,China)

机构地区：[1]首都医科大学细胞生物学系,北京100069 [2]‘肝脏保护和再生调节’北京市重点实验室,北京100069

出　　处：《中国细胞生物学学报》2018年第11期1886-1893,共8页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81430013)资助的课题~~

摘　　要：该文采用实时荧光定量聚合酶链反应(RT-qPCR)、蛋白印记实验(Western blot)、流式细胞分析(flurescence-activated cell sorting, FACS)技术检测骨髓单核巨噬细胞(bone marrow monocytes/macrophages, BMMs)中大麻素受体1(cannabinoid receptor 1, CB1)对BMMs极化的作用。结果表明,使用1μmol/L ACEA(CB1激动剂)处理细胞发现, BMMs中M1型的标志物CD86、IL-1、MIP-1β、NOS2、IL-6、TNF-α的m RNA水平均上调;用流式细胞分析技术检测发现, BMMs中M1型的标志物CD86蛋白质水平上调;使用PI3K/AKT信号通路的特异性抑制剂(LY294002)预处理BMMs,再加入1μmol/L ACEA刺激细胞,与未加入LY294002的对照组相比,这些M1型BMMs标志物的mRNA表达均被抑制;通过Western blot法证实ACEA使p-AKT增加,而使用1μmol/L AM281(CB1药理学拮抗剂)阻断CB1功能,则抑制了ACEA导致的p-AKT增加。上述结果说明, CB1通过PI3K/AKT信号通路介导BMMs向M1型极化。RT-qPCR, Western blot, flurescence-activated cell sorting(FACS) were employed to detect whether cannabinoid receptor 1(CB1) was involved in bone marrow monocytes/macrophage(BMMs) M1 polarization. The results showed that ACEA(CB1 agonist, 1 μmol/L) promoted the mRNA levels of M1 type macrophage gene signatures(CD86, IL-1, MIP-1β, NOS2, IL-6, TNF-α) in BMMs. The protein level of CD86 deteced by FACS were increased in BMMs induced by ACEA. When BMMs were pretreated with LY294002(specific PI3 K/AKT signal pathway inhibitor), ACEA-induced(1 μmol/L) increases of M1 gene signatures mRNA levels was suppressed. Furthermore, Western blot analysis showed the protein level of phosphorylated AKT(p-AKT) was increased in ACEA-treated BMMs. When BMMs were pretreated with AM281(CB1 antagonist, 1 μmol/L), p-AKT protein level was inhibited. The result showed that CB1 mediated monocyte/macrophage M1 polarization via PI3 K/AKT signaling pathway.

关 键 词：大麻素受体1 单核巨噬细胞极化 PI3K/AKT信号通路 

分 类 号：R285[医药卫生—中药学]