Limits...
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 is a novel thymic immunosuppressive pentapeptide originally obtained from calf thymic immunosuppressive extract.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.

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

Effects of TIPP on the activation of mitogen-activated protein (MAP) kinases (A–D) and extracellular signal-regulated protein kinase (ERK) kinase 1/2 (MEK1/2) (E,F) in IgE-antigen complex stimulated RBL-2H3 cells. The Western blot diagram is a representative of three independent experiment diagrams with similar results. Each lane was loaded with 20 μg of total protein. Results are expressed as mean ± SEM (n = 3). Compared with Normal group, ##p < 0.01, ###p < 0.001; compared with Control group, *p < 0.05, **p < 0.01.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4307361&req=5

ijms-16-02252-f008: Effects of TIPP on the activation of mitogen-activated protein (MAP) kinases (A–D) and extracellular signal-regulated protein kinase (ERK) kinase 1/2 (MEK1/2) (E,F) in IgE-antigen complex stimulated RBL-2H3 cells. The Western blot diagram is a representative of three independent experiment diagrams with similar results. Each lane was loaded with 20 μg of total protein. Results are expressed as mean ± SEM (n = 3). Compared with Normal group, ##p < 0.01, ###p < 0.001; compared with Control group, *p < 0.05, **p < 0.01.

Mentions: Mitogen-activated protein (MAP) kinase (extracellular signal-regulated protein kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 mitogen-activated protein kinase (p38)) pathways have been identified to reflect RBL-2H3 activation. Thus, the levels of phosphorylation of three MAP kinases were examined. The results showed that the antigen stimulation caused apparent increases in the phosphorylation of the three MAP kinases compared with normal cells. TIPP treatment significantly reduced the upregulation of phosphorylation of ERK in activated RBL-2H3 cells but showed no effect on the levels of phosphorylation of p38 and JNK (Figure 8A–D).


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)

Effects of TIPP on the activation of mitogen-activated protein (MAP) kinases (A–D) and extracellular signal-regulated protein kinase (ERK) kinase 1/2 (MEK1/2) (E,F) in IgE-antigen complex stimulated RBL-2H3 cells. The Western blot diagram is a representative of three independent experiment diagrams with similar results. Each lane was loaded with 20 μg of total protein. Results are expressed as mean ± SEM (n = 3). Compared with Normal group, ##p < 0.01, ###p < 0.001; compared with Control group, *p < 0.05, **p < 0.01.
© Copyright Policy
Related In: Results  -  Collection

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

ijms-16-02252-f008: Effects of TIPP on the activation of mitogen-activated protein (MAP) kinases (A–D) and extracellular signal-regulated protein kinase (ERK) kinase 1/2 (MEK1/2) (E,F) in IgE-antigen complex stimulated RBL-2H3 cells. The Western blot diagram is a representative of three independent experiment diagrams with similar results. Each lane was loaded with 20 μg of total protein. Results are expressed as mean ± SEM (n = 3). Compared with Normal group, ##p < 0.01, ###p < 0.001; compared with Control group, *p < 0.05, **p < 0.01.
Mentions: Mitogen-activated protein (MAP) kinase (extracellular signal-regulated protein kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 mitogen-activated protein kinase (p38)) pathways have been identified to reflect RBL-2H3 activation. Thus, the levels of phosphorylation of three MAP kinases were examined. The results showed that the antigen stimulation caused apparent increases in the phosphorylation of the three MAP kinases compared with normal cells. TIPP treatment significantly reduced the upregulation of phosphorylation of ERK in activated RBL-2H3 cells but showed no effect on the levels of phosphorylation of p38 and JNK (Figure 8A–D).

Bottom Line: TIPP is a novel thymic immunosuppressive pentapeptide originally obtained from calf thymic immunosuppressive extract.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.

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