PI3K/Akt pathway is involved in the activation of RAW 264.7 cells induced by hydroxypropyltrimethyl ammonium chloride ch
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PI3K/Akt pathway is involved in the activation of RAW 264.7 cells induced by hydroxypropyltrimethyl ammonium chloride chitosan* YANG Yue1, 2, 3, XING Rong’e1, 2, **, LIU Song1, 2, QIN Yukun1, 2, LI Kecheng1, 2, YU Huahua1, 2, LI Pengcheng1, 2, ** 1
Key Laboratory of Experimental Marine Biology, Center for Ocean Mega-Science, Institute of Oceanology, Chinese Academy
2
Laboratory for Marine Drugs and Bioproducts of Qingdao National Laboratory for Marine Science and Technology, Qingdao
3
University of Chinese Academy of Sciences, Beijing 100049, China
of Sciences, Qingdao, China 266237, China
Received Jan. 19, 2019; accepted in principle Jun. 28, 2019; accepted for publication Sep. 8, 2019 © Chinese Society for Oceanology and Limnology, Science Press and Springer-Verlag GmbH Germany, part of Springer Nature 2019
Abstract We previously demonstrated that 2-hydroxypropyltrimethyl ammonium chloride chitosan (HACC) promoted the production of nitric oxide (NO) and proinflammatory cytokines by activating the mitogen-activated protein kinases (MAPK) and Janus kinase (JAK)/STAT pathways in RAW 264.7 cells, indicating good immunomodulatory activity of HACC. In this study, to further investigate the immunomodulatory mechanisms of HACC, we determined the roles of phosphatidylinositol 3-kinase (PI3K)/Akt, activating protein (AP-1) and nuclear factor kappa B (NF-κB) in HACC-induced activation of RAW 264.7 cells by the western blotting. The results suggest that HACC promoted the phosphorylation of p85 and Akt. Furthermore, c-Jun and p65 were also increased after the treatment of RAW 264.7 cells with HACC, indicating the translocation of NF-κB and AP-1 from cytoplasm to nucleus. In addition, as scanning electron microscopy (SEM) analysis shows, the cell morphology changed after HACC treatment. These findings indicate that HACC activated MAPK, JAK/STAT, and PI3K/Akt signaling pathways dependent on AP-1 and NF-κB activation in RAW 264.7 cells, ultimately leading to the increase of NO and cytokines. Keyword: hydroxypropyltrimethyl ammonium chloride chitosan; RAW 264.7 cells; PI3K/Akt pathway; nuclear factor-κB; activating protein 1
1 INTRODUCTION Immunity plays vital role in vertebrates. The immune system can broadly be divided into two branches: innate immunity and adaptive immunity (Fang and Zhang, 2016). Macrophages along with dendritic cells are two important innate immune system members. RAW 264.7 cells, a kind of macrophage derived from mouse ascites, are widely utilized as in vitro model to detect immunomodulatory effect of foreign substances. Upon activation, RAW 264.7 cells secrete cytokines through the activation of signaling pathways (Sun et al., 2015). The signaling pathways of RAW 264.7 cells have been elucidated by many researchers, among them, phosphoinositide 3-kinases (PI3K)/Akt, sarcoma (Src) family kinases,
mitogen-activated protein kinase (MAPK) including p38, extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), Janus kinase/signal transducer and activator of transcription (JAK-STAT), an
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