Comparative study on modeccin- and phytohemagglutinin (PHA)-induced secretion of cytokines and nitric oxide (NO) in RAW264.7 cells

Kim, Daekyung; Yamasaki, Yasuhiro; Jiang, Zedong; Nakayama, Yuki; Yamanishi, Tomohiro; Yamaguchi, Kensei; Oda, Tatsuya

doi:10.1093/abbs/gmq105

Cited by 16 publications

(12 citation statements)

References 40 publications

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“…To investigate whether or not MAP kinase signal pathways are involved in NO-production in RAW264.7 cells stimulated by ascophyllan and fucoidans, the effects of PD98059, SB202190 and SP600125, which are specific inhibitors for ERK, p38, and JNK MAP kinase, respectively, were examined at final 10 M of each inhibitor and total 24 h incubation. Under the conditions, the inhibitors themselves showed no cytotoxic effect on RAW264.7 cells [39]. As shown in Table 1, SP600125 showed a potent inhibitory effect on ascophyllan-induced NO production in RAW264.7 cells, while PD98059 and SB202190 were less effective.…”

Section: Effects Of Three Map Kinase Inhibitors On No-inducing Activimentioning

confidence: 91%

“…To estimate NO level in RAW264.7 cells, nitrite, a stable reaction product of NO with molecular oxygen, was measured by Griess assay as described previously [39]. In brief, adherent RAW264.7 cells in 96-well plates (3 x 10 4 cells/well) were treated with varying concentrations of polysaccharide samples (0-200 g/ml) for 18 h in the growth medium at 37℃, and then the nitrite levels in the culture medium were measured.…”

Section: Nitrite Assay For the Estimation Of Nitric Oxide (No)mentioning

confidence: 99%

“…Enzyme-linked immunosorbent assay (ELISA) Adherent RAW264.7 cells in 96-well plates (3 x 10 4 cells/well) were treated with varying concentrations of polysaccharide samples (0 to 200 g/ml) in the growth medium at 37℃. After 18 h incubation, the levels of TNF- and G-CSF in the culture supernatantsof treated cells were measured by sandwich ELISA with two antibodies to two different epitopes on TNF- or G-CSF molecule as described previously[39]. The TNF- and G-CSF concentrations were estimated from a reference to a standard curve for serial two-fold dilution of murine recombinant TNF- or G-CSF.Electrophoretic mobility shift assay (EMSA)Adherent RAW264.7 cells (2 x 10 6 cells/3.5 cm dish) were treated with 100 g/ml of polysaccharide samples in serum-free DMEM at 37℃.…”

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The potent activity of sulfated polysaccharide, ascophyllan, isolated from Ascophyllum nodosum to induce nitric oxide and cytokine production from mouse macrophage RAW264.7 cells: Comparison between ascophyllan and fucoidan

Jiang

Okimura²,

Yamaguchi

et al. 2011

Nitric Oxide

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Ascophyllan isolated from the brown alga Ascophyllum nodosum is a fucose-containing sulfated polysaccharide, which has similar but distinct characteristic monosaccharide composition and entire chemical structure to fucoidan. In this study, we examined the effects of ascophyllan, fucoidan isolated from A. nodosum (A-fucoidan), and fucoidan from Sigma (S-fucoidan) as a representative fucoidan derived from other source (Fucus vesiculosus) on mouse macrophage cell line RAW264.7 cells. No significant cytotoxic effects of ascophyllan and A-fucoidan on RAW264.7 cells were observed up to 1000μg/ml, while S-fucoidan showed cytotoxic effect in a concentration-dependent manner. Ascophyllan induced extremely higher level of nitric oxide (NO) production from RAW264.7 cells than those induced by fucoidans over the concentration range tested (0-200μg/ml). Reverse transcription polymerase chain reaction (RT-PCR) and western blot analysis revealed that expression level of inducible NO synthase (iNOS) in ascophyllan-treated RAW264.7 cells was much higher than the levels detected in the cells treated with fucoidans. Furthermore, the activities of ascophyllan to induce the secretion of tumor necrosis factor-α (TNF-α) and granulocyte colony-stimulating factor (G-CSF) from RAW264.7 cells were also greater than those induced by fucoidans especially at lower concentration range (3.1-50μg/ml). The activities of ascophyllan to induce NO and cytokine production in mouse peritoneal macrophages were also stronger than those of fucoidans. Electrophoretic mobility shift assay (EMSA) using infrared dye labeled nuclear factor-kappa B (NF-κB) and AP-1 consensus sequences suggested that ascophyllan can strongly activate these transcription factors. Marked increase in the nuclear translocation of p65, and the phosphorylation and degradation of IκB-α were also observed in ascophyllan-treated RAW264.7 cells. Analysis using mitogen-activated protein (MAP) kinase inhibitors and western blot analysis suggested that c-Jun N-terminal kinase (JNK) and p38 MAP kinase are mainly involved in ascophyllan-induced NO production.

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Section: Effects Of Three Map Kinase Inhibitors On No-inducing Activimentioning

confidence: 91%

Section: Nitrite Assay For the Estimation Of Nitric Oxide (No)mentioning

confidence: 99%

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confidence: 99%

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The potent activity of sulfated polysaccharide, ascophyllan, isolated from Ascophyllum nodosum to induce nitric oxide and cytokine production from mouse macrophage RAW264.7 cells: Comparison between ascophyllan and fucoidan

Jiang

Okimura²,

Yamaguchi

et al. 2011

Nitric Oxide

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“…RAW264.7 mouse macrophage cell line has long been used as an in vitro model to study inflammatory molecules synthetic response to a wide variety of stimuli (Shin, 2008;Jeong, 2009). It is known that macrophages produce multiple inflammatory molecules such as NO, TNF-α, IL-1, IL-12, interferon-g and chemokines in response to the stimulation of some lectins (Kesherwani et al, 2007), such as phytohemagglutinin to stimulate RAW264.7 cells could induce the production of NO (Daekyung et al, 2011). Nitric oxide is a multifunctional biomolecule involved in a variety of physiological and pathological processes, such as vascular regulation, host immune defense, neurotransmission and other systems (Paige, 2007).…”

Section: Discussionmentioning

confidence: 99%

Effects of forsythoside on lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages

Guan¹,

Shen²,

Zhang³

et al. 2013

Afr. J. Pharm. Pharmacol.

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“…Furthermore, PHA has been used to improve the efficiency of the nuclear transfer of somatic cells for oocytes from diverse species 19, because its capacity to induce closer contacts between adjacent cell membranes. In addition, PHA can stimulate the production of nitric oxide (NO) in different cells 20,21. Some studies have shown that neuronal NO synthase (nNOS) is expressed in ovarian follicles 22 and is involved in the control of folliculogenesis, steroidogenesis, oocyte maturation, and ovulation through the production of NO 23.…”

Section: Introductionmentioning

confidence: 99%

Phytohemagglutinin improves the development and ultrastructure of in vitro-cultured goat (Capra hircus) preantral follicles

Cunha

Costa

Rossi

et al. 2013

Braz J Med Biol Res

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The objective this study was to determine the effect of phytohemagglutinin (PHA) on survival, growth and gene expression in caprine secondary follicles cultured in vitro. Secondary follicles (∼0.2 mm) were isolated from the cortex of caprine ovaries and cultured individually for 6 days in α-MEM+ supplemented with PHA (0, 1, 10, 50, 100, or 200 µg/mL). After 6 days of culture, follicle diameter and survival, antrum formation, ultrastructure and expression of mRNA for FSH receptors (FSH-R), proliferating cell nuclear antigen (PCNA), and neuronal nitric oxide synthase were determined. All treatments maintained follicular survival [α-MEM+ (94.59%); 1 µg/mL PHA (96.43%); 10 µg/mL PHA (84.85%); 50 µg/mL PHA (85.29%); 100 µg/mL PHA (88.57%), and 200 µg/mL PHA (87.50)], but the presence of 10 µg/mL PHA in the culture medium increased the antrum formation rate (21.21%) when compared with control (5.41%, P < 0.05) and ensured the maintenance of oocyte and granulosa cell ultrastructures after 6 days of culture. The expression of mRNA for FSH-R (2.7 ± 0.1) and PCNA (4.4 ± 0.2) was also significantly increased in follicles cultured with 10 µg/mL PHA in relation to those cultured in α-MEM+ (1.0 ± 0.1). In conclusion, supplementation of culture medium with 10 µg/mL PHA maintains the follicular viability and ultrastructure, and promotes the formation of antral cavity after 6 days of culture in vitro.

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Comparative study on modeccin- and phytohemagglutinin (PHA)-induced secretion of cytokines and nitric oxide (NO) in RAW264.7 cells

Cited by 16 publications

References 40 publications

The potent activity of sulfated polysaccharide, ascophyllan, isolated from Ascophyllum nodosum to induce nitric oxide and cytokine production from mouse macrophage RAW264.7 cells: Comparison between ascophyllan and fucoidan

The potent activity of sulfated polysaccharide, ascophyllan, isolated from Ascophyllum nodosum to induce nitric oxide and cytokine production from mouse macrophage RAW264.7 cells: Comparison between ascophyllan and fucoidan

Effects of forsythoside on lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages

Phytohemagglutinin improves the development and ultrastructure of in vitro-cultured goat (Capra hircus) preantral follicles

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