Protein Expression Changes of Cellular Proliferation-Related Proteins in Murine Macrophages, RAW 264.7 Cells by Ddialyzed Coffee Extract Treatment

2017 ◽  
Vol 41 (2) ◽  
pp. 63-71 ◽  
Author(s):  
Cheol Soo Yoon ◽  
◽  
Suk Keun Lee
Keyword(s):  
Protein Expression ◽  
Cellular Proliferation ◽  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Murine Macrophages ◽  
Related Proteins

scholarly journals 4-hexylresorcinol-induced protein expression changes in human umbilical cord vein endothelial cells as determined by immunoprecipitation high-performance liquid chromatography

PLoS ONE ◽  
2020 ◽  
Vol 15 (12) ◽  
pp. e0243975
Author(s):  
Yeon Sook Kim ◽  
Dae Won Kim ◽  
Seong-Gon Kim ◽  
Suk Keun Lee
Keyword(s):  
Wound Healing ◽  
Endothelial Cells ◽  
Protein Expression ◽  
Macrophage Polarization ◽  
Raw 264.7 Cells ◽  
Human Umbilical Cord ◽  
Raw 264.7 ◽  
Related Proteins ◽  
Umbilical Cord Vein ◽  
Human Umbilical Cord Vein

4-Hexylresorcinol (4HR) is used as a food preservative and an ingredient of toothpaste and cosmetics. The present study was performed using 233 antisera to determine the changes in protein expression induced by 4HR in human umbilical cord vein endothelial cells (HUVECs), and evaluated the 4HR-induced effects in comparison with previous results (Kim et al., 2019). Similar to RAW 264.7 cells, 4HR-treated HUVECs showed decreases in the expression of the proliferation-related proteins, cMyc/MAX/MAD network proteins, p53/RB and Wnt/β-catenin signaling, and they showed inactivation of DNA transcription and protein translation compared to the untreated controls. 4HR upregulated growth factors (TGF-β1, β2, β3, SMAD2/3, SMAD4, HGF-α, Met, IGF-1) and RAS signaling proteins (RAF-B, p38, p-p38, p-ERK-1, and Rab-1), and induced stronger expression of the cellular protection-, survival-, and differentiation-related proteins in HUVECs than in RAW 264.7 cells. 4HR suppressed NFkB signaling in a manner that suggests potential anti-inflammatory and wound healing effects by reducing M1 macrophage polarization and increasing M2 macrophage polarization in both cells. 4HR-treated HUVECs tended to increase the ER stress mediators by upregulating eIF2AK3, ATF4, ATF6, lysozyme, and LC3 and downregulating eIF2α and GADD153 (CHOP), resulting in PARP-1/AIF-mediated apoptosis. These results indicate that 4HR has similar effects on the protein expression of HUVECs and RAW 264.7 cells, but their protein expression levels differ according to cell types. The 4HR-treated cells showed global protein expression characteristic of anticancer and wound healing effects, which could be alleviated simultaneously by other proteins exerting opposite functions. These results suggest that although 4HR has similar effects on the global protein expression of HUVECs and RAW 264.7 cells, the 4HR-induced molecular interferences in those cells are complex enough to produce variable protein expression, leading different cell functions. Moreover, HUVECs have stronger wound healing potential to overcome the impact induced by 4HR than RAW 264.7 cells.

Anti-inflammatory effect of chitosan oligosaccharides in RAW 264.7 cells

2010 ◽  
Vol 5 (1) ◽  
pp. 95-102 ◽  
Author(s):  
Eun-Jin Yang ◽  
Jong-Gwan Kim ◽  
Ji-Young Kim ◽  
Seong Kim ◽  
Nam Lee ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Protein Expression ◽  
Inducible Nitric Oxide Synthase ◽  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Chitosan Oligosaccharides ◽  
Cox 2 ◽  
Oxide Synthase ◽  
Inducible Nitric Oxide

AbstractWe examined the effects of chitosan oligosaccharides (COSs) with different molecular weights (COS-A, 10 kDa < MW < 20 kDa; COS-C, 1 kDa < MW < 3 kDa) on the lipopolysaccharide (LPS)-induced production of prostaglandin E2 and nitric oxide and on the expression of cyclooxygenase-2 and inducible nitric oxide synthase in RAW264.7 macrophages. COS-A (0.4%) and COS-C (0.2%) significantly inhibited PGE2 production in LPS-stimulated macrophages without cytotoxicity. The effect of COS-A and COS-C on COX-2 expression in activated macrophages was also investigated by immunoblotting. The inhibition of PGE2 by COS-A and COS-C can be attributed to the blocking of COX-2 protein expression. COS-A (0.4%) and COS-C (0.2%) also markedly inhibited the LPS-induced NO production of RAW 264.7 cells by 50.2% and 44.1%, respectively. The inhibition of NO by COSs was consistent with decreases in inducible nitric oxide synthase (iNOS) protein expression. To test the inhibitory effects of COS-A and COS-C on other cytokines, we also performed ELISA assays for IL-1β in LPS-stimulated RAW 264.7 macrophage cells, but only a dose-dependent decrease in the IL-1β production exerted by COS-A was observed. In order to test for irritation and the potential sensitization of COS-A and COS-C for use as cosmetic materials, human skin primary irritation tests were performed on 32 volunteers; no adverse reactions of COSs usage were observed. Based on these results, we suggest that COS-A and COS-C be considered possible anti-inflammatory candidates for topical application.

Prompt detection of endogenous hypochlorite (ClO−) in murine macrophages and zebrafish embryos facilitated by a distinctive chemodosimetric mode

2020 ◽  
Vol 18 (34) ◽  
pp. 6716-6723
Author(s):  
Shrabani Saha ◽  
Sujoy Das ◽  
Sriparna Das ◽  
Anwesha Samanta ◽  
Sudipta Maitra ◽  
...  
Keyword(s):  
Danio Rerio ◽  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Zebrafish Embryos ◽  
Murine Macrophages ◽  
Naphthalene Diimide

A fluorescein appended naphthalene diimide based probe (FANDI) has been developed to selectively recognize exogenous as well as endogenous ClO− ions in RAW 264.7 cells (macrophages) and zebrafish embryos (Danio rerio).

scholarly journals Burkholderia pseudomallei Type III Secretion System Mutants Exhibit Delayed Vacuolar Escape Phenotypes in RAW 264.7 Murine Macrophages

2008 ◽  
Vol 76 (7) ◽  
pp. 2991-3000 ◽  
Author(s):  
Mary N. Burtnick ◽  
Paul J. Brett ◽  
Vinod Nair ◽  
Jonathan M. Warawa ◽  
Donald E. Woods ◽  
...  
Keyword(s):  
Type Iii Secretion ◽  
Burkholderia Pseudomallei ◽  
Type Iii Secretion System ◽  
Secretion System ◽  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Triple Mutant ◽  
Necrosis Factor Alpha ◽  
Murine Macrophages ◽  
Type Iii

ABSTRACT Burkholderia pseudomallei is a facultative intracellular pathogen capable of surviving and replicating within eukaryotic cells. Recent studies have shown that B. pseudomallei Bsa type III secretion system 3 (T3SS-3) mutants exhibit vacuolar escape and replication defects in J774.2 murine macrophages. In the present study, we characterized the interactions of a B. pseudomallei bsaZ mutant with RAW 264.7 murine macrophages. Following uptake, the mutant was found to survive and replicate within infected RAW 264.7 cells over an 18-h period. In addition, high levels of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), granulocyte-macrophage colony-stimulating factor (GM-CSF), and RANTES, but not IL-1α and IL-1β, were detected in culture supernatants harvested from infected monolayers. The subcellular location of B. pseudomallei within infected RAW 264.7 cells was determined, and as expected, the bsaZ mutant demonstrated early-vacuolar-escape defects. Interestingly, however, experiments also indicated that this mutant was capable of delayed vacuolar escape. Consistent with this finding, evidence of actin-based motility and multinucleated giant cell formation were observed between 12 and 18 h postinfection. Further studies demonstrated that a triple mutant defective in all three B. pseudomallei T3SSs exhibited the same phenotype as the bsaZ mutant, indicating that functional T3SS-1 and T3SS-2 did not appear to be responsible for the delayed escape phenotype in RAW 264.7 cells. Based upon these findings, it appears that B. pseudomallei may not require T3SS-1, -2, and -3 to facilitate survival, delayed vacuolar escape, and actin-based motility in activated RAW 264.7 macrophages.

scholarly journals Extensive protein expression changes induced by pamidronate in RAW 264.7 cells as determined by IP-HPLC

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e9202 ◽  
Author(s):  
Sang Shin Lee ◽  
Soung Min Kim ◽  
Yeon Sook Kim ◽  
Suk Keun Lee
Keyword(s):  
Epigenetic Modification ◽  
Protein Translation ◽  
Raw 264.7 Cells ◽  
Proliferation Index ◽  
Ras Signaling ◽  
Therapeutic Effects ◽  
Raw 264.7 ◽  
Protein Expressions ◽  
Related Proteins ◽  
In Cells

Background Bisphosphonate therapy has become a popular treatment for osteoporosis, Paget’s disease, multiple myeloma, osteogenesis imperfecta, myocardial infarction, and cancer despite its serious side effects. Bisphosphonate-induced molecular signaling changes in cells are still not clearly elucidated. Methods As bisphosphonates are primarily engulfed by macrophages, we treated RAW 264.7 cells (a murine macrophage cell line) with pamidronate and investigated global protein expressional changes in cells by immunoprecipitation high performance liquid chromatography (IP-HPLC) using 218 antisera. Results Pamidronate upregulated proliferation-activating proteins associated with p53/Rb/E2F and Wnt/β-catenin pathways, but downregulated the downstream of RAS signaling, pAKT1/2/3, ERK-1, and p-ERK-1, and subsequently suppressed cMyc/MAX/MAD network. However, in situ proliferation index of pamidronate-treated RAW264.7 cells was slightly increased by 3.2% vs. non-treated controls. Pamidronate-treated cells showed increase in the expressions of histone- and DNA methylation-related proteins but decrease of protein translation-related proteins. NFkB signaling was also suppressed as indicated by the down-regulations of p38 and p-p38 and the up-regulation of mTOR, while the protein expressions related to cellular protection, HSP-70, NRF2, JNK-1, and LC3 were upregulated. Consequently, pamidronate downregulated the protein expressions related to immediate inflammation,cellular differentiation, survival, angiogenesis, and osteoclastogenesis, but upregulated PARP-1 and FAS-mediated apoptosis proteins. These observations suggest pamidronate affects global protein expressions in RAW 264.7 cells by stimulating cellular proliferation, protection, and apoptosis but suppressing immediate inflammation, differentiation, osteoclastogenesis, and angiogenesis. Accordingly, pamidronate appears to affect macrophages in several ways eliciting not only its therapeutic effects but also atypical epigenetic modification, protein translation, RAS and NFkB signalings. Therefore, our observations suggest pamidronate-induced protein expressions are dynamic, and the affected proteins should be monitored by IP-HPLC to achieve the therapeutic goals during treatment.

Sign in / Sign up

Export Citation Format

Share Document