Chemical characterization of a novel polysaccharide ASKP-1 from Artemisia sphaerocephala Krasch seed and its macrophage activation via MAPK, PI3k/Akt and NF-κB signaling pathways in RAW264.7 cells

The aim of this study was to investigate the molecular mechanism underlying the immunomodulatory effect of the purified Artemisia sphaerocephala Krasch seed polysaccharide (ASKP-1) in RAW264.7 macrophages.

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Characterization of polysaccharide from longan pulp as the macrophage stimulator

RSC Advances ◽

10.1039/c5ra16044h ◽

2015 ◽

Vol 5 (118) ◽

pp. 97163-97170 ◽

Cited By ~ 21

Author(s):

Yang Yi ◽

Hongxun Wang ◽

Ruifen Zhang ◽

Ting Min ◽

Fei Huang ◽

...

Keyword(s):

Signaling Pathways ◽

P38 Mapk ◽

Macrophage Activation

A 44.7-kDa polysaccharide LPIIa from longan pulp was mainly composed of →6)-Glc-(1→, →5)-Ara-(1→, →4)-Man-(1→ and →6)-Gal-(1→. It stimulated macrophage activation partlyviaTLR4 and TLR2, followed by p38 MAPK- and NF-κB-dependent signaling pathways.

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Frutescone O from Baeckea frutescens Blocked TLR4-Mediated Myd88/NF-κB and MAPK Signaling Pathways in LPS Induced RAW264.7 Macrophages

Frontiers in Pharmacology ◽

10.3389/fphar.2021.643188 ◽

2021 ◽

Vol 12 ◽

Author(s):

Xiaobing Lin ◽

Junhan Zhang ◽

Decai Fan ◽

Jiqin Hou ◽

Hao Wang ◽

...

Keyword(s):

Signaling Pathways ◽

Molecular Mechanisms ◽

Mapk Signaling ◽

Raw264.7 Cells ◽

Raw264.7 Macrophages ◽

Anti Inflammatory ◽

Baeckea Frutescens ◽

Myd88 Signaling ◽

Myeloid Differentiation Factor ◽

Mapk Signaling Pathways

Frutescone O was isolated from the aerial parts of Baeckea frutescens L., which was commonly used as a folk medicinal material for treating anti-inflammatory disease in South East Asia. This study aimed to investigate the anti-inflammatory activity and related signaling cascade of Frutescone O (Fru) in LPS induced RAW264.7 cells. The anti-inflammation activity of Frutescone O was determined according to the inhibitory effects on the secretion of nitric oxide (NO), expression of inducible NO synthase, and pro-inflammatory cytokines. The regulation of Myeloid differentiation factor 88 (Myd88), inhibition of NF-κB, and MAPK pathways were further investigated for molecular mechanisms. Fru significantly decreased the expression of iNOS and the production of NO in LPS-stimulated RAW264.7 cells. It also dose-dependently suppressed LPS induced expression of IL-1β, IL-6, and TNF-α. Furthermore, Fru remarkably inhibited the upregulation of NF-κB (p50) expression in the nucleus and the phosphorylation ratio of p38, JNK, ERK, and Myd88 signaling protein. The molecular docking and cellular thermal shift assay (CETSA) results indicated that Fru participated in a robust and stable interaction with the active site of TLR4-MD2. Thus, Fru suppressed the LPS induced inflammation in RAW264.7 cells by blocking the TLR4 mediated signal transduction through the NF-κB and MAPK signaling pathways and inhibiting the Myd88 and iNOS expression.

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The immunomodulatory effect of docosahexaenoic acid (DHA) on the RAW264.7 cells by modification of the membrane structure and function

Food & Function ◽

10.1039/c9fo02618e ◽

2020 ◽

Vol 11 (3) ◽

pp. 2603-2616 ◽

Cited By ~ 3

Author(s):

Nana Bie ◽

Lirong Han ◽

Meng Meng ◽

Zhongli Yan ◽

Chunling Wang

Keyword(s):

Docosahexaenoic Acid ◽

Molecular Mechanism ◽

Membrane Structure ◽

Structure And Function ◽

Immunomodulatory Effect ◽

Raw264.7 Cells ◽

Immunomodulatory Activity ◽

Physiological Functions ◽

Membrane Structure And Function ◽

And Function

DHA can regulate various physiological functions of cells. Our group has clarified the immunomodulatory activity and molecular mechanism of DHA on RAW264.7 cells.

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Structural characterization of arabinogalactan extracted from Ixeris chinensis (Thunb.) Nakai and its immunomodulatory effect on RAW264.7 macrophages

International Journal of Biological Macromolecules ◽

10.1016/j.ijbiomac.2019.09.158 ◽

2020 ◽

Vol 143 ◽

pp. 977-983

Author(s):

Hongyan Li ◽

Wancui Xie ◽

Xing Qiao ◽

Huanhuan Cui ◽

Xihong Yang ◽

...

Keyword(s):

Structural Characterization ◽

Immunomodulatory Effect ◽

Raw264.7 Macrophages

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The Functional Characterization of Phosphorylation of Tristetraprolin at C-Terminal NOT1-binding Domain

10.21203/rs.3.rs-50794/v1 ◽

2020 ◽

Author(s):

Hsin-Hui Hsieh ◽

Yen-An Chen ◽

Yao-Jen Chang ◽

Hsin-Hui Wang ◽

Ya-Han Yu ◽

...

Keyword(s):

Functional Characterization ◽

Binding Domain ◽

Raw264.7 Cells ◽

Mrna Stabilization ◽

Raw264.7 Macrophages ◽

Phosphatase 2A ◽

Mrna Destabilization ◽

Ribosomal S6 Kinase ◽

Tandem Ccch Zinc Finger

Abstract Backgound: Tristetraprolin (TTP) family proteins contain conserved tandem CCCH zinc-finger binding to AU-rich elements and C-terminal NOT1-binding domain. TTP is phosphorylated extensively in cells and its mRNA destabilization activity is regulated by protein phosphorylation. Methods: We generated an antibody against phospho-Serine 316 located at C-terminal NOT1-binding site and examined TTP phosphorylation in LPS-stimulated RAW264.7 cells. Knockout of TTP in RAW264.7 cells using CRISPR/Cas9 gene editing was created to explore TTP functions. Results: We demonstrated that Ser316 was phosphorylated by p90 ribosomal S6 kinase 1 (RSK1) and p38-activated protein kinase (MK2), and dephosphorylated by Protein Phosphatase 2A (PP2A). Phosphorylation-mimic mutant of S316D resulted in dissociation with CCR4-NOT deadenylase complex through weakening interaction with CNOT1. Furthermore, Ser316 and serines 52 and 178 were independently contributed to CCR4-NOT complex recruitment in the immunoprecipitation assay using phosphor-mimic mutants. In RAW264.7 macrophages, TTP was induced and Ser316 was phosphorylated through RSK1 and MK2 by LPS stimulation. Knockout of TTP resulted in TNFα mRNA increased due to mRNA stabilization. Overexpression of non-phosphorylated S316A TTP mutant can restore TTP activity and lead to TNFα mRNA decreased. GST pull-down and RNA pull-down analyses demonstrated that endogenous TTP with Ser316 phosphorylation decreased the interaction with CNOT1. Conclusions: Our results suggest that the TTP-mediated mRNA stability is modulated by Ser316 phosphorylation to regulate the TTP interaction with CCR4-NOT deadenylase complex.

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Bioactivity-based analysis and chemical characterization of anti-inflammatory compounds from Curcuma zedoaria rhizomes using LPS-stimulated RAW264.7 cells

Bioorganic Chemistry ◽

10.1016/j.bioorg.2018.09.027 ◽

2019 ◽

Vol 82 ◽

pp. 26-32 ◽

Cited By ~ 9

Author(s):

Tae Kyoung Lee ◽

Tuy An Trinh ◽

Seoung Rak Lee ◽

Sil Kim ◽

Hae Min So ◽

...

Keyword(s):

Chemical Characterization ◽

Raw264.7 Cells ◽

Curcuma Zedoaria ◽

Anti Inflammatory

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The functional characterization of phosphorylation of tristetraprolin at C-terminal NOT1-binding domain

Journal of Inflammation ◽

10.1186/s12950-021-00288-2 ◽

2021 ◽

Vol 18 (1) ◽

Author(s):

Hsin-Hui Hsieh ◽

Yen-An Chen ◽

Yao-Jen Chang ◽

Hsin-Hui Wang ◽

Ya-Han Yu ◽

...

Keyword(s):

Functional Characterization ◽

Binding Domain ◽

Raw264.7 Cells ◽

Mrna Stabilization ◽

Raw264.7 Macrophages ◽

Phosphatase 2A ◽

Mrna Destabilization ◽

Ribosomal S6 Kinase ◽

Tandem Ccch Zinc Finger

Abstract Background Tristetraprolin (TTP) family proteins contain conserved tandem CCCH zinc-finger binding to AU-rich elements and C-terminal NOT1-binding domain. TTP is phosphorylated extensively in cells, and its mRNA destabilization activity is regulated by protein phosphorylation. Methods We generated an antibody against phospho-Serine316 located at the C-terminal NOT1-binding site and examined TTP phosphorylation in LPS-stimulated RAW264.7 cells. Knockout of TTP was created in RAW264.7 cells using CRISPR/Cas9 gene editing to explore TTP functions. Results We demonstrated that Ser316 was phosphorylated by p90 ribosomal S6 kinase 1 (RSK1) and p38-activated protein kinase (MK2) and dephosphorylated by Protein Phosphatase 2A (PP2A). A phosphorylation-mimic mutant of S316D resulted in dissociation with the CCR4-NOT deadenylase complex through weakening interaction with CNOT1. Furthermore, Ser316 and serines 52 and 178 were independently contributed to the CCR4-NOT complex recruitment in the immunoprecipitation assay using phosphor-mimic mutants. In RAW264.7 macrophages, TTP was induced, and Ser316 was phosphorylated through RSK1 and MK2 by LPS stimulation. Knockout of TTP resulted in TNFα mRNA increased due to mRNA stabilization. Overexpression of non-phosphorylated S316A TTP mutant can restore TTP activity and lead to TNFα mRNA decreased. GST pull-down and RNA pull-down analyses demonstrated that endogenous TTP with Ser316 phosphorylation decreased the interaction with CNOT1. Conclusions Our results suggest that the TTP-mediated mRNA stability is modulated by Ser316 phosphorylation via regulating the TTP interaction with the CCR4-NOT deadenylase complex.

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