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Inhibition of the IgE-mediated activation of RBL-2H3 cells by TIPP, a novel thymic immunosuppressive pentapeptide.

Lian Q, Cheng Y, Zhong C, Wang F - Int J Mol Sci (2015)

Bottom Line: TIPP significantly suppressed the increase of intracellular calcium and the rearrangement of F-actin, attenuated the transcription of pro-inflammatory cytokines (IL-3, -4, -6, -13, TNF-α, and monocyte chemotactic protein-1 (MCP-1)), and decreased the expression of COX-2.Western blot analysis showed that TIPP had an inhibitory activity on the phosphorylation of extracellular signal-regulated protein kinase 1/2 (ERK1/2) and ERK kinase 1/2 (MEK1/2), and inhibited the activation of NF-κB.The data suggested that TIPP effectively suppressed IgE-mediated activation of RBL-2H3 cells via blocking MEK/ERK and NF-κB signaling pathways.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Chemical Biology of Natural Products (Ministry of Education), Institute of Biochemical and Biotechnological Drugs, School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China. xhrg_1042@126.com.

ABSTRACT
TIPP is a novel thymic immunosuppressive pentapeptide originally obtained from calf thymic immunosuppressive extract. The present study aimed to investigate the inhibitory activity of TIPP on IgE-mediated activation of RBL-2H3 cells. Release of β-hexosaminidase and histamine, intracellular calcium, membrane ruffling, mRNA levels of cytokines, cyclooxygenase-2 (COX-2) expression, and activation of mitogen-activated protein kinases (MAP kinases) and NF-κB were determined by colorimetric assay, fluorescence spectrophotometer, confocal fluorescence microscope, quantification PCR, and Western blot, respectively. The results showed that TIPP significantly inhibited the degranulation in IgE-antigen complex-stimulated RBL-2H3 cells without cytotoxicity. TIPP significantly suppressed the increase of intracellular calcium and the rearrangement of F-actin, attenuated the transcription of pro-inflammatory cytokines (IL-3, -4, -6, -13, TNF-α, and monocyte chemotactic protein-1 (MCP-1)), and decreased the expression of COX-2. Western blot analysis showed that TIPP had an inhibitory activity on the phosphorylation of extracellular signal-regulated protein kinase 1/2 (ERK1/2) and ERK kinase 1/2 (MEK1/2), and inhibited the activation of NF-κB. The data suggested that TIPP effectively suppressed IgE-mediated activation of RBL-2H3 cells via blocking MEK/ERK and NF-κB signaling pathways.

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Effect of TIPP on intracellular calcium level. This result is representative of three independent experiments with similar results.
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ijms-16-02252-f003: Effect of TIPP on intracellular calcium level. This result is representative of three independent experiments with similar results.

Mentions: The inhibitory effect of TIPP on the intracellular calcium level of activated RBL-2H3 cells was detected with Fluo 3-AM ((4-(6-acetoxymethoxy-2,7-dichloro-3-oxo-9-xanthenyl)-4'-methyl-2,2'(ethylenedioxy)dianiline-N,N,N',N'-tetraacetic acid tetrakis(acetoxymethyl) ester)), a calcium-specific fluorescent probe. As shown in Figure 3, antigen stimulation on sensitized RBL-2H3 cells caused an increase in the fluorescence intensity changes, which indicated that the intracellular calcium level was increased by antigen stimulation. TIPP (200 and 800 μg/mL) and ketotifen pretreatments could suppress the intracellular calcium level changes caused by antigen as the results showed decreased changes in fluorescence intensity in treated groups.


Inhibition of the IgE-mediated activation of RBL-2H3 cells by TIPP, a novel thymic immunosuppressive pentapeptide.

Lian Q, Cheng Y, Zhong C, Wang F - Int J Mol Sci (2015)

Effect of TIPP on intracellular calcium level. This result is representative of three independent experiments with similar results.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4307361&req=5

ijms-16-02252-f003: Effect of TIPP on intracellular calcium level. This result is representative of three independent experiments with similar results.
Mentions: The inhibitory effect of TIPP on the intracellular calcium level of activated RBL-2H3 cells was detected with Fluo 3-AM ((4-(6-acetoxymethoxy-2,7-dichloro-3-oxo-9-xanthenyl)-4'-methyl-2,2'(ethylenedioxy)dianiline-N,N,N',N'-tetraacetic acid tetrakis(acetoxymethyl) ester)), a calcium-specific fluorescent probe. As shown in Figure 3, antigen stimulation on sensitized RBL-2H3 cells caused an increase in the fluorescence intensity changes, which indicated that the intracellular calcium level was increased by antigen stimulation. TIPP (200 and 800 μg/mL) and ketotifen pretreatments could suppress the intracellular calcium level changes caused by antigen as the results showed decreased changes in fluorescence intensity in treated groups.

Bottom Line: TIPP significantly suppressed the increase of intracellular calcium and the rearrangement of F-actin, attenuated the transcription of pro-inflammatory cytokines (IL-3, -4, -6, -13, TNF-α, and monocyte chemotactic protein-1 (MCP-1)), and decreased the expression of COX-2.Western blot analysis showed that TIPP had an inhibitory activity on the phosphorylation of extracellular signal-regulated protein kinase 1/2 (ERK1/2) and ERK kinase 1/2 (MEK1/2), and inhibited the activation of NF-κB.The data suggested that TIPP effectively suppressed IgE-mediated activation of RBL-2H3 cells via blocking MEK/ERK and NF-κB signaling pathways.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Chemical Biology of Natural Products (Ministry of Education), Institute of Biochemical and Biotechnological Drugs, School of Pharmaceutical Sciences, Shandong University, Jinan 250012, China. xhrg_1042@126.com.

ABSTRACT
TIPP is a novel thymic immunosuppressive pentapeptide originally obtained from calf thymic immunosuppressive extract. The present study aimed to investigate the inhibitory activity of TIPP on IgE-mediated activation of RBL-2H3 cells. Release of β-hexosaminidase and histamine, intracellular calcium, membrane ruffling, mRNA levels of cytokines, cyclooxygenase-2 (COX-2) expression, and activation of mitogen-activated protein kinases (MAP kinases) and NF-κB were determined by colorimetric assay, fluorescence spectrophotometer, confocal fluorescence microscope, quantification PCR, and Western blot, respectively. The results showed that TIPP significantly inhibited the degranulation in IgE-antigen complex-stimulated RBL-2H3 cells without cytotoxicity. TIPP significantly suppressed the increase of intracellular calcium and the rearrangement of F-actin, attenuated the transcription of pro-inflammatory cytokines (IL-3, -4, -6, -13, TNF-α, and monocyte chemotactic protein-1 (MCP-1)), and decreased the expression of COX-2. Western blot analysis showed that TIPP had an inhibitory activity on the phosphorylation of extracellular signal-regulated protein kinase 1/2 (ERK1/2) and ERK kinase 1/2 (MEK1/2), and inhibited the activation of NF-κB. The data suggested that TIPP effectively suppressed IgE-mediated activation of RBL-2H3 cells via blocking MEK/ERK and NF-κB signaling pathways.

Show MeSH
Related in: MedlinePlus