Activation of RAW264.7 mouse macrophage cells in vitro through treatment with recombinant ricin toxin-binding subunit B: Involvement of protein tyrosine, NF-κB and JAK-STAT kinase signaling pathways

ABSTRACT Salmonella enterica encodes a wide array of virulence factors. One novel virulence factor, an A2B5 toxin known as the typhoid toxin (TT), was recently identified among a variety of S. enterica serovars. While past studies have shown that some serovars encode both the TT (active subunits CdtB and PltA and binding subunit PltB) and a second binding subunit (ArtB), these serovars were thought to be the exception. Here, we show that genes encoding the TT are detected in more than 100 serovars representing distinct phylogenetic lineages of S. enterica subsp. enterica, although clade B and section Typhi are significantly more likely to encode TT genes than serovars from other clades. Furthermore, we show that 81% of these TT-positive serovars also encode artB, suggesting that the cooccurrence of both toxin binding subunits is considerably more common than previously thought. A combination of in silico modeling, bacterial two-hybrid system screening, and tandem affinity purification (TAP) of toxin subunits suggests that ArtB and PltB interact in vitro, at least under some growth conditions. While different growth conditions yielded slightly higher transcript abundances of artB and pltB, both genes had their highest relative transcript abundances when Salmonella was grown under low-Mg2+ conditions, suggesting that ArtB and PltB may compete for inclusion in the TT. Together, our results suggest that ArtB likely plays an important and previously underappreciated role in the biology of the TT produced by typhoidal and nontyphoidal Salmonella. IMPORTANCE While previous reports had suggested that the typhoid toxin (TT) could potentially use ArtB as an alternate binding subunit, this was thought to play a minor role in the evolution and biology of the toxin. In this study, we establish that both TT genes and artB are widespread among Salmonella enterica subsp. enterica, suggesting that TT likely plays a broader role in Salmonella virulence that extends beyond its proposed role in typhoid fever. Furthermore, our data suggest the selective maintenance of both toxin binding subunits, which may compete for inclusion in the holotoxin. Last, our data support the importance of characterizing diverse nontyphoidal Salmonella (NTS) serovars, as the presence of classically defined typhoidal virulence factors among NTS serovars continues to challenge the typhoid-nontyphoid Salmonella paradigm.

Download Full-text

Curcumin and Curcumol Inhibit NF-κB and TGF-β1/Smads Signaling Pathways in CSE-Treated RAW246.7 Cells

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2019/3035125 ◽

2019 ◽

Vol 2019 ◽

pp. 1-9

Author(s):

Ning Li ◽

Tian-Hao Liu ◽

Jing-Ze Yu ◽

Chen-Xi Li ◽

Yang Liu ◽

...

Keyword(s):

Signaling Pathways ◽

Signaling Pathway ◽

Treated Group ◽

Macrophage Cells ◽

Injury Model ◽

Intracellular Ros ◽

Anti Inflammatory ◽

Main Components ◽

Inflammatory Effect

E-Zhu (Curcuma zedoaria) is known as a classical traditional Chinese medicine and widely used in the treatment of cancers, cardiovascular disease, inflammation, and other diseases. Its main components include curcumol and curcumin, which have anti-inflammatory and antifibrosis effects. Here we established an in vitro inflammatory injury model by stimulating RAW246.7 cells with cigarette smoke extract (CSE) and detected the intervention effects of curcumin and curcumol on CSE-treated Raw246.7 macrophage cells to explore whether the two compounds inhibited the expression of inflammatory cytokines by inhibiting the NF-κB signaling pathway. We detected the antifibrosis effects of curcumin and curcumol via TGF-β1/Smads signaling pathways. The model of macrophage damage group was established by CSE stimulation. Curcumol and curcumin were administered to Raw246.7 macrophage cells. The efficacy of curcumol and curcumin was evaluated by comparing the activation of proinflammatory factors, profibrotic factors, and NF-κB and TGF-β1/Smads signaling pathway. In addition, CSE-treated group was employed to detect whether the efficacy of curcumol and curcumin was dependent on the NF-κB signaling via the pretreatment with the inhibitor of NF-κB. Our findings demonstrated that curcumol and curcumin could reduce the release of intracellular ROS from macrophages, inhibit the NF-κB signaling pathway, and downregulate the release of proinflammatory factor. Curcumol and curcumin inhibited the TGF-β1/Smads signaling pathway and downregulated the release of fibrotic factors. Curcumin showed no anti-inflammatory effect in CSE-treated cells after the inhibition of NF-κB. Curcumol and curcumin showed an anti-inflammatory effect by inhibiting the NF-κB signaling pathway.

Download Full-text

CircEpc1 Promotes Ricin Toxin-Induced Inflammation via Activation of NF-κB and MAPK Signaling Pathways by Sponging miR-5114

Frontiers in Pharmacology ◽

10.3389/fphar.2021.767900 ◽

2021 ◽

Vol 12 ◽

Author(s):

Mingxin Dong ◽

Xiaohao Zhang ◽

Haotian Yu ◽

Yan Wang ◽

Ying Chang ◽

...

Keyword(s):

Signaling Pathways ◽

Suppressive Effect ◽

Mapk Signaling ◽

Innate Immune ◽

Circular Rnas ◽

Inflammatory Injury ◽

Ricin Toxin ◽

Mapk Signaling Pathways

Increasing studies have concentrated on investigating circular RNAs (circRNAs) as pivotal regulators in the progression of numerous diseases and biological processes and abundant evidence shows that circRNAs are participated in the regulation of innate immune responses. Several studies showed that Ricin Toxin (RT) could induce inflammatory injury. There was no research on the particular functions and underlying mechanisms of circRNAs in RT-induced inflammation. In this study, RNA sequencing performed on RT-treated and normal RAW264.7 macrophage cells was used to investigated the differentially expressed circRNAs. Based on the dataset, the expression of circEpc1 (mmu_circ_0,000,842) was identified higher in RT-treated cells. Moreover, gain-and-loss function assays showed that circEpc1 function as a promoter in RT-induced inflammation in vivo and in vitro. Mechanistically, circEpc1 acted as a miR-5114 sponge to relieve the suppressive effect of miR-5114 on its target NOD2 and thereby activating NF-κB and MAPK signaling pathways. Our results illuminated a link between RT-induced inflammation and the circEpc1 regulatory loop and provided novel insight into the functions of circRNA in innate immune, which may emerge as a potential target in immunotherapy to control the RT-induced inflammatory injury.

Download Full-text

CircEpc1 Promotes Ricin Toxin-Induced Inflammation Via Activation of NF-κB And MAPK Signaling Pathways By Sponging miR-5114

10.21203/rs.3.rs-786451/v1 ◽

2021 ◽

Author(s):

Mingxin Dong ◽

Xiaohao Zhang ◽

Haotian Yu ◽

Yan Wang ◽

Ying Chang ◽

...

Keyword(s):

Signaling Pathways ◽

Suppressive Effect ◽

Mapk Signaling ◽

Innate Immune ◽

Circular Rnas ◽

Inflammatory Injury ◽

Ricin Toxin ◽

Mapk Signaling Pathways

Abstract Background: Increasing studies have concentrated on investigating circular RNAs (circRNAs) as pivotal regulators in the progression of numerous diseases and biological processes and abundant evidence shows that circRNAs are participated in the regulation of innate immune responses. Several studies showed that Ricin Toxin (RT) could induce inflammatory injury. There was no research on the particular functions and underlying mechanisms of circRNAs in RT-induced inflammation. Results: In this study, RNA sequencing performed on RT-treated and normal RAW264.7 macrophage cells was used to investigated the differentially expressed circRNAs. Based on the dataset, the expression of circEpc1 (mmu_circ_0000842) was identified higher in RT-treated cells. Moreover, gain-and-loss function assays showed that circEpc1 function as a promoter in RT-induced inflammation in vivo and in vitro. Mechanistically, circEpc1 acted as a miR-5114 sponge to relieve the suppressive effect of miR-5114 on its target NOD2 and thereby activating NF-κB and MAPK signaling pathways. Conclusions: Our results illuminated a link between RT-induced inflammation and the circEpc1 regulatory loop and provided novel insight into the functions of circRNA in innate immune, which may emerge as a potential target in immunotherapy to control the RT-induced inflammatory injury.

Download Full-text

Polygoni Rhizoma Inhibits Inflammatory Response through Inactivation of Nuclear Factor-kappaB and Mitogen Activated Protein Kinase Signaling Pathways in RAW264.7 Mouse Macrophage Cells

Phytotherapy Research ◽

10.1002/ptr.3530 ◽

2011 ◽

Vol 26 (2) ◽

pp. 239-245 ◽

Cited By ~ 13

Author(s):

Hyuk-Jin Seo ◽

Jeong-Eun Huh ◽

Jong-Hyun Han ◽

Soo-Jin Jeong ◽

Jinsil Jang ◽

...

Keyword(s):

Protein Kinase ◽

Inflammatory Response ◽

Signaling Pathways ◽

Mitogen Activated Protein Kinase ◽

Nuclear Factor Kappab ◽

Nuclear Factor ◽

Mouse Macrophage ◽

Kinase Signaling ◽

Mitogen Activated Protein ◽

Protein Kinase Signaling

Download Full-text

Synthesis of N-acyl derivatives of Pro-Gly-Pro-Leu peptide: Proteolytic stability in vitro and effects on mouse macrophage cells RAW264.7

Doklady Biochemistry and Biophysics ◽

10.1134/s1607672917050118 ◽

2017 ◽

Vol 476 (1) ◽

pp. 333-336

Author(s):

K. V. Shevchenko ◽

V. V. Bezuglov ◽

M. G. Akimov ◽

I. Yu. Nagaev ◽

V. P. Shevchenko ◽

...

Keyword(s):

Mouse Macrophage ◽

Macrophage Cells ◽

Proteolytic Stability ◽

Derivatives Of ◽

Acyl Derivatives

Download Full-text

In Vitro Effects of Cetylated Fatty Acids Mixture from Celadrin on Chondrogenesis and Inflammation with Impact on Osteoarthritis

Cartilage ◽

10.1177/1947603518775798 ◽

2018 ◽

Vol 11 (1) ◽

pp. 88-97 ◽

Cited By ~ 3

Author(s):

Ariana Hudita ◽

Bianca Galateanu ◽

Sorina Dinescu ◽

Marieta Costache ◽

Anca Dinischiotu ◽

...

Keyword(s):

Fatty Acids ◽

Stem Cells ◽

Cellular Level ◽

Animal Origin ◽

Mouse Macrophage ◽

3 Dimensional ◽

Macrophage Cells ◽

Culture Model ◽

In Vitro Effects

Objective Cetylated fatty acids are a group of naturally occurring fats of plant and/or animal origin. Cetyl myristoleate, in particular, was initially involved in osteoarthritis related research as its therapeutic administration prevented experimentally induced arthritis in Swiss Albino mice. In this context, the aim of our study was to investigate the possible mechanisms of Celadrin cetylated fatty acids action at the cellular level inflammation related pain relief and chondrogenesis. Design For this, we tested the effects of the cetylated fatty acids mixture from Celadrin on an in vitro scaffold-free 3-dimensional mesenchymal stem cells culture model of chondrogenesis. Furthermore, we treated stimulated mouse macrophage cells with the cetylated fatty acids mixture to investigate the expression profile of secreted inflammatory cytokines. Results The cetylated fatty acids mixture from Celadrin significantly decreased the production of IL-6, MCP-1, and TNF, key regulators of the inflammatory process, in stimulated RAW264.7 mouse macrophage cells. The treatment with cetylated fatty acids mixture initiated and propagated the process of chondrogenesis as demonstrated by the increased expression and deposition of chondrogenic markers by the differentiating mesenchymal cells. Conclusion The cetylated fatty acids mixture from Celadrin reduces inflammation in vitro by significantly decreasing the expression of IL-6, MCP-1, and TNF in stimulated RAW264.7 mouse macrophage cells. These compounds facilitate the chondrogenic differentiation process of human adipose-derived stem cells by stimulating the expression of chondrogenic markers under chondrogenic induction conditions.

Download Full-text