scholarly journals Pentoxifylline-induced Protein Expression Change in RAW 264.7 Cells as Determined by Immunoprecipitation-based High Performance Liquid Chromatography

2021 ◽  
Author(s):  
Suk Keun Lee ◽  
Mi Hyun Seo ◽  
Dae Won Kim ◽  
Yeon Sook Kim
Keyword(s):  
High Performance Liquid Chromatography ◽  
Er Stress ◽  
High Performance ◽  
Low Dose ◽  
Epigenetic Modification ◽  
Osteoblastic Differentiation ◽  
Raw 264.7 Cells ◽  
High Dose ◽  
Cell Mediated Immunity ◽  
Raw 264.7

Although pentoxifylline (PTX) was identified as a competitive non-selective phosphodiesterase inhibitor, its pharmacological effect has not been clearly elucidated. The present study explored the effect of low dose 10 μg/mL PTX (therapeutic dose) compared to high dose 300 μg/mL PTX (experimental dose) in RAW 264.7 cells through immunoprecipitation-based high performance liquid chromatography (IP-HPLC), immunohistochemistry, and western blot. 10 μg/mL PTX increased the expression of proliferation (Ki-67, PCNA, cyclin D2, cdc25A), epigenetic modification (KDM4D, PCAF), protein translation (DOHH, DHPS, eIF5A1), RAS signaling (KRAS, pAKT1/2/3, PI3K), NFkB signaling (NFkB, GADD45, p38), protection (HSP70, SOD1, GSTO1/2), neuromuscular differentiation (NSEγ, myosin-1a, desmin), osteoblastic differentiation (BMP2, RUNX2, osterix), acute inflammation (TNFα, IL-1, CXCR4), innate immunity (β-defensin 1, lactoferrin, TLR-3, -4), cell-mediated immunity (CD4, CD8, CD80), while decreased the expression of ER stress (eIF2α, eIF2AK3, ATF6α), fibrosis (FGF2, CTGF, collagen 3A1), and chronic inflammation (CD68, MMP-2, -3, COX2) versus the untreated controls. 10 μg/mL PTX enhanced FAS-mediated apoptosis but diminished p53-mediated apoptosis, and downregulated many angiogenesis proteins (angiogenin, VEGF-A, and FLT4), but upregulated HIF1α, VEGFR2, and CMG2 reactively. Whereas, 300 μg/mL PTX consistently decreased proliferation, epigenetic modification, RAS and NFkB signaling, neuromuscular and osteoblastic differentiation, but increased apoptosis, ER stress, and fibrosis compared to 10 μg/mL PTX. These data suggest PTX has different biological effect on RWA 264.7 cells depending on the concentration of 10 μg/mL and 300 μg/mL PTX. The low dose 10 μg/mL PTX enhanced RAS/NFkB signaling, proliferation, differentiation, and inflammation, particularly, it stimulated neuromuscular and osteoblastic differentiation, innate immunity, and cell-mediated immunity, but attenuated ER stress, fibrosis, angiogenesis, and chronic inflammation, while the high dose 300 μg/mL PTX was found to alleviate the 10 μg/mL PTX-induced biological effects, resulted in the suppression of RAS/NFkB signaling, proliferation, neuromuscular and osteoblastic differentiation, and inflammation.

scholarly journals In VitroActivities of Daptomycin-, Vancomycin-, and Teicoplanin-Loaded Polymethylmethacrylate against Methicillin-Susceptible, Methicillin-Resistant, and Vancomycin-Intermediate Strains of Staphylococcus aureus

2011 ◽  
Vol 55 (12) ◽  
pp. 5480-5484 ◽  
Author(s):  
Yuhan Chang ◽  
Wen-Chien Chen ◽  
Pang-Hsin Hsieh ◽  
Dave W. Chen ◽  
Mel S. Lee ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
High Performance ◽  
Low Dose ◽  
Antibacterial Activities ◽  
High Dose ◽  
Bone Cements ◽  
Antibacterial Effects ◽  
Methicillin Resistant ◽  
Content Type

ABSTRACTThe objective of this study was to evaluate the antibacterial effects of polymethylmethacrylate (PMMA) bone cements loaded with daptomycin, vancomycin, and teicoplanin against methicillin-susceptibleStaphylococcus aureus(MSSA), methicillin-resistantStaphylococcus aureus(MRSA), and vancomycin-intermediateStaphylococcus aureus(VISA) strains. Standardized cement specimens made from 40 g PMMA loaded with 1 g (low-dose), 4 g (middle-dose) or 8 g (high-dose) antibiotics were tested for elution characteristics and antibacterial activities. The patterns of release of antibiotics from the cement specimens were evaluated usingin vitrobroth elution assay with high-performance liquid chromatography. The activities of broth elution fluid against differentStaphylococcus aureusstrains (MSSA, MRSA, and VISA) were then determined. The antibacterial activities of all the tested antibiotics were maintained after being mixed with PMMA. The cements loaded with higher dosages of antibiotics showed longer elution periods. Regardless of the antibiotic loading dose, the teicoplanin-loaded cements showed better elution efficacy and provided longer inhibitory periods against MSSA, MRSA, and VISA than cements loaded with the same dose of vancomycin or daptomycin. Regarding the choice of antibiotics for cement loading in the treatment ofStaphylococcus aureusinfection, teicoplanin was superior in terms of antibacterial effects.

Sa1779 Vitamin D Modulates T Cell-Mediated Immunity: Results From a Randomized Controlled Trial of Low-Dose and High-Dose Vitamin D3

2014 ◽  
Vol 146 (5) ◽  
pp. S-294 ◽  
Author(s):  
Gauree G. Konijeti ◽  
Matthew R. Boylan ◽  
Yanna Song ◽  
Pankaj Arora ◽  
Frank E. Harrell ◽  
...  
Keyword(s):  
Vitamin D ◽  
Randomized Controlled Trial ◽  
T Cell ◽  
Vitamin D3 ◽  
Low Dose ◽  
Controlled Trial ◽  
High Dose ◽  
Cell Mediated Immunity ◽  
Randomized Controlled ◽  
High Dose Vitamin

Fermentation Improves Anti-Inflammatory Effect of Sipjeondaebotang on LPS-Stimulated RAW 264.7 Cells

2012 ◽  
Vol 40 (04) ◽  
pp. 813-831 ◽  
Author(s):  
You-Chang Oh ◽  
Won-Kyung Cho ◽  
Yun Hee Jeong ◽  
Ga Young Im ◽  
Min Cheol Yang ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Inflammatory Mediators ◽  
High Performance ◽  
Biological Effects ◽  
Inhibitory Effect ◽  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Tnf Α ◽  
Cox 2 ◽  
Anti Inflammatory

Sipjeondaebotang (SJ) has been used as a traditional drug in east-Asian countries. In this study, to provide insight into the biological effects of SJ and SJ fermented by Lactobacillus, we investigated their effects on lipopolysaccharide (LPS)-mediated inflammation in macrophages. The investigation was focused on whether SJ and fermented SJ could inhibit the production of pro-inflammatory mediators such as prostaglandin (PG) E2 and nitric oxide (NO) as well as the expressions of cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS), tumor necrosis factor (TNF)-α, mitogen-activated protein kinases (MAPKs) and nuclear factor (NF)-κB in LPS-stimulated RAW 264.7 cells. We found that SJ modestly inhibited LPS-induced PGE2, NO and TNF-α production as well as the expressions of COX-2 and iNOS. Interestingly, fermentation significantly increased its inhibitory effect on the expression of all pro-inflammatory mediators. Furthermore, fermented SJ exhibited increased inhibition of p38 MAPK and c-Jun NH2-terminal kinase (JNK) MAPK phosphorylation as well as NF-κB p65 translocation by reduced IκBα degradation compared with either untreated controls or unfermented SJ. High performance liquid chromatography (HPLC) analysis showed fermentation by Lactobacillus increases liquiritigenin and cinnamyl alcohol contained in SJ, which are known for their anti-inflammatory activities. Finally, SJ fermented by Lactobacillus exerted potent anti-inflammatory activity by inhibiting MAPK and NF-κB signaling in RAW 264.7 cells.

scholarly journals A Combination Extract of Gardeniae Fructus and Perillae Folium Exerts Anti-Inflammatory Effects on LPS-Activated RAW 264.7 Mouse Macrophages via an ER Stress-Induced CHOP Pathway

Processes ◽  
2021 ◽  
Vol 9 (9) ◽  
pp. 1632
Author(s):  
Wansu Park
Keyword(s):  
Er Stress ◽  
P38 Mapk ◽  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Mrna Levels ◽  
Mapk Phosphorylation ◽  
Inflammatory Reactions ◽  
Inflammatory Protein ◽  
Mouse Macrophages ◽  
Anti Inflammatory

The aim of this study is to investigate the effects of a combination extract of Gardeniae Fructus and Perillae Folium (GP) on inflammatory reactions in lipopolysaccharide (LPS)-activated mouse macrophages RAW 264.7 cells. Multiplex cytokine assay, Fluo-4 calcium assay, Flow cytometry assay for phospho-P38 MAPK, and quantitative PCR were carried out. GP significantly reduced LPS-induced productions of macrophage inflammatory protein (MIP)-1α and monokine induced by gamma interferon (MIG) and release of intracellular calcium in LPS-activated RAW 264.7 cells. GP also significantly inhibited P38 MAPK phosphorylation and mRNA levels of Chop, Camk2a, Stat1, Stat3, Jak2, Fas, Nos2, and Ptgs2 in LPS-activated RAW 264.7 cells. Taken together, this study represents that GP exerts anti-inflammatory effects on LPS-activated RAW 264.7 cells via ER stress-induced CHOP pathway.

scholarly journals Role of the Inflammatory Response of RAW 264.7 Cells in the Metastasis of Novel Cancer Stem-Like Cells

Medicina ◽  
2021 ◽  
Vol 57 (8) ◽  
pp. 778
Author(s):  
Chan-Yen Kuo ◽  
Tzu-Hsien Yang ◽  
Pei-Fang Tsai ◽  
Chun-Hsien Yu
Keyword(s):  
Er Stress ◽  
Tumor Progression ◽  
Conditioned Medium ◽  
Inflammatory Responses ◽  
Macrophage Polarization ◽  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Ros Accumulation ◽  
Oxide Synthase ◽  
Cells Cultured

Background and objectives: Tumor progression and the immune response are intricately linked. Additionally, the presence of macrophages in the microenvironment is essential for carcinogenesis, but regulation of the polarization of M1- and M2-like macrophages and their role in metastasis remain unclear. Based on previous studies, both reactive oxygen species (ROS) and the endoplasmic reticulum (ER) are emerging as key players in macrophage polarization. While it is known that cancers alter macrophage inflammatory responses to promote tumor progression, there is limited knowledge regarding how they affect the macrophage-dependent innate host defense. Materials and methods: We detected the levels of ROS, the ability of chemotaxis, the expressions of markers of M1-/M2-like macrophages in RAW264.7 in presence of T2- and T2C-conditioned medium. Results: The results of this study indicated that ROS levels were decreased in RAW 264.7 cells when cultured with T2C-conditioned medium, while there was an improvement in chemotaxis abilities. We also found that the M2-like macrophages were characterized by an elongated shape in RAW 264.7 cells cultured in T2C-conditioned medium, which had increased CD206 expression but decreased expression of CD86 and inducible nitric oxide synthase. Suppression of ER stress shifted polarized M1-like macrophages toward an M2-like phenotype in RAW 264.7 cells cultured in T2C-conditioned medium. Conclusions: Taken together, we conclude that the polarization of macrophages is associated with the alteration of cell shape, ROS accumulation, and ER stress.

scholarly journals Proinflammatory Cytokine Gene Expression by Murine Macrophages in Response toBrugia malayi WolbachiaSurface Protein

2007 ◽  
Vol 2007 ◽  
pp. 1-7 ◽  
Author(s):  
Chantima Porksakorn ◽  
Surang Nuchprayoon ◽  
Kiwon Park ◽  
Alan L. Scott
Keyword(s):  
Inflammatory Responses ◽  
Surface Protein ◽  
Protein Molecule ◽  
Innate Immune ◽  
Proteinase K ◽  
Cytokine Gene Expression ◽  
Raw 264.7 Cells ◽  
High Dose ◽  
Raw 264.7 ◽  
Proinflammatory Response

Wolbachia, an endosymbiotic bacterium found in most species of filarial parasites, is thought to play a significant role in inducing innate inflammatory responses in lymphatic filariasis patients. However, theWolbachia-derived molecules that are recognized by the innate immune system have not yet been identified. In this study, we exposed the murine macrophage cell line RAW 264.7 to a recombinant form of the majorWolbachiasurface protein (rWSP) to determine if WSP is capable of innately inducing cytokine transcription. Interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF) mRNAs were all upregulated by the rWSP stimulation in a dose-dependant manner. TNF transcription peaked at 3 hours, whereas IL-1βand IL-6 transcription peaked at 6 hours post-rWSP exposure. The levels of innate cytokine expression induced by a high-dose (9.0μg/mL) rWSP in the RAW 264.7 cells were comparable to the levels induced by 0.1μg/mLE. coli-derived lipopolysaccharides. Pretreatment of the rWSP with proteinase-K drastically reduced IL-1β, IL-6, and TNF transcription. However, the proinflammatory response was not inhibited by polymyxin B treatment. These results strongly suggest that the majorWolbachiasurface protein molecule WSP is an important inducer of innate immune responses during filarial infections.

scholarly journals Effect of prochloraz fungicide on neopterin and biopterin concentrations in blood plasma of common carp

2014 ◽  
Vol 83 (2) ◽  
pp. 101-105 ◽  
Author(s):  
Petr Maršálek ◽  
Ivana Mikuliková ◽  
Helena Modrá ◽  
Zdeňka Svobodová
Keyword(s):  
High Performance Liquid Chromatography ◽  
Blood Plasma ◽  
Common Carp ◽  
High Performance ◽  
Pesticide Exposure ◽  
Cell Mediated Immunity ◽  
Traditional Role ◽  
Reverse Phase ◽  
Exposed Fish ◽  
In Blood Plasma

Neopterin and biopterin are often used as markers of cell mediated immunity. The aim of this study was to evaluate the effect of prochloraz on neopterin and biopterin concentrations in blood plasma of common carp as possible indicators for monitoring perturbations of the immune system caused by xenobiotics like pesticides. Prochloraz is a widely used imidazole fungicide in horticulture and agriculture. A total of 60 juvenile common carp were divided into four groups of 15 fish and exposed to prochloraz at concentrations of 0, 50, 150 and 380 µg·l-1, respectively. The total length of exposure was 28 days. Neopterin and biopterin concentrations were determined by reverse phase high performance liquid chromatography with fluorescence detection. Our results indicated that concentrations of neopterin (25 ± 7.6 nmol·l-1) and biopterin (190 ± 29 nmol·l-1) in plasma of untreated common carp were comparable with those in mammals. Neopterin concentrations significantly (P< 0.01) increased after exposure to prochloraz in comparison to non-exposed fish, while biopterin concentrations were not influenced by exposure to prochloraz. This study contains new data on neopterin and biopterin concentrations in fish plasma and investigates neopterin and biopterin in their non-traditional role as markers of cell mediated immunity of fish associated with pesticide exposure.

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