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Effect of mitogen-activated protein kinases on chemokine synthesis induced by substance P in mouse pancreatic acinar cells.

Ramnath RD, Sun J, Adhikari S, Bhatia M - J. Cell. Mol. Med. (2007 Nov-Dec)

Bottom Line: Moreover, substance P-induced ERK 1/2, JNK, NFkappaB and AP-1 activation as well as chemokine synthesis were blocked by pre-treatment with either extracellular signal-regulated protein kinase kinase 1 (MEK1) inhibitor or JNK inhibitor.In addition, substance P-induced activation of ERK 112, JNK, NFkappaB and AP-1-driven chemokine production were attenuated by CP96345, a selective NK1 R antagonist, in pancreatic acinar cells.Taken together, these results suggest that substance P-NK1 R induced chemokine production depends on the activation of MAPKs-mediated NFkappaB and AP-1 signalling pathways in mouse pancreatic acini.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, National University of Singapore, Singapore.

ABSTRACT
Substance P, acting via its neurokinin 1 receptor (NK1 R), plays an important role in mediating a variety of inflammatory processes. Its interaction with chemokines is known to play a crucial role in the pathogenesis of acute pancreatitis. In pancreatic acinar cells, substance P stimulates the release of NFkappaB-driven chemokines. However, the signal transduction pathways by which substance P-NK1 R interaction induces chemokine production are still unclear. To that end, we went on to examine the participation of mitogen-activated protein kinases (MAPKs) in substance P-induced synthesis of pro-inflammatory chemokines, monocyte chemoanractant protein-1 (MCP-I), macrophage inflammatory protein-lalpha (MIP-lalpha) and macrophage inflammatory protein-2 (MIP-2), in pancreatic acini. In this study, we observed a time-dependent activation of ERK1/2, c-Jun N-terminal kinase (JNK), NFkappaB and activator protein-1 (AP-1) when pancreatic acini were stimulated with substance P. Moreover, substance P-induced ERK 1/2, JNK, NFkappaB and AP-1 activation as well as chemokine synthesis were blocked by pre-treatment with either extracellular signal-regulated protein kinase kinase 1 (MEK1) inhibitor or JNK inhibitor. In addition, substance P-induced activation of ERK 112, JNK, NFkappaB and AP-1-driven chemokine production were attenuated by CP96345, a selective NK1 R antagonist, in pancreatic acinar cells. Taken together, these results suggest that substance P-NK1 R induced chemokine production depends on the activation of MAPKs-mediated NFkappaB and AP-1 signalling pathways in mouse pancreatic acini.

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Substance P-NK1R interaction is involved in ERK1/2 and JNK activation. Freshly isolated pancreatic acini, obtained from three mice, were pre-incubated with 1 μM CP96345 for half an hour at 37°C followed by stimulation with 1 μM substance P for 45 min for ERK1/2 and JNK at 37°C. Cells were subsequently lysed, and cell proteins were subjected to Western blot analysis using antibodies against (A) phospho-ERK, total ERK1/2 (B) phospho-JNK, total JNK. Corresponding densitometric analysis of Western blot experiments from pancreatic acini. The results are representative of three independent experiments. Results shown are the means + SE. * P≤ 0.05 when compared to control, + P ≤0.05 when compared to substance P (SP).
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fig08: Substance P-NK1R interaction is involved in ERK1/2 and JNK activation. Freshly isolated pancreatic acini, obtained from three mice, were pre-incubated with 1 μM CP96345 for half an hour at 37°C followed by stimulation with 1 μM substance P for 45 min for ERK1/2 and JNK at 37°C. Cells were subsequently lysed, and cell proteins were subjected to Western blot analysis using antibodies against (A) phospho-ERK, total ERK1/2 (B) phospho-JNK, total JNK. Corresponding densitometric analysis of Western blot experiments from pancreatic acini. The results are representative of three independent experiments. Results shown are the means + SE. * P≤ 0.05 when compared to control, + P ≤0.05 when compared to substance P (SP).

Mentions: Our data show that substance P-induced ERK1/2 and JNK activation were mediated through NK1R. We pre-treated the pancreatic acini with 1 μM of CP96345, a selective NK1R antagonist, followed by stimulation with 1 μM of substance P for 45 min. Cells were then lysed, and cell proteins were subjected to Western blot analysis. Our results, in Figure 8, demonstrate that CP96345 significantly reduced substance P-induced ERK1/2 and JNK activation in pancreatic acinar cells when compared to substance P-only treated cells.


Effect of mitogen-activated protein kinases on chemokine synthesis induced by substance P in mouse pancreatic acinar cells.

Ramnath RD, Sun J, Adhikari S, Bhatia M - J. Cell. Mol. Med. (2007 Nov-Dec)

Substance P-NK1R interaction is involved in ERK1/2 and JNK activation. Freshly isolated pancreatic acini, obtained from three mice, were pre-incubated with 1 μM CP96345 for half an hour at 37°C followed by stimulation with 1 μM substance P for 45 min for ERK1/2 and JNK at 37°C. Cells were subsequently lysed, and cell proteins were subjected to Western blot analysis using antibodies against (A) phospho-ERK, total ERK1/2 (B) phospho-JNK, total JNK. Corresponding densitometric analysis of Western blot experiments from pancreatic acini. The results are representative of three independent experiments. Results shown are the means + SE. * P≤ 0.05 when compared to control, + P ≤0.05 when compared to substance P (SP).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4401295&req=5

fig08: Substance P-NK1R interaction is involved in ERK1/2 and JNK activation. Freshly isolated pancreatic acini, obtained from three mice, were pre-incubated with 1 μM CP96345 for half an hour at 37°C followed by stimulation with 1 μM substance P for 45 min for ERK1/2 and JNK at 37°C. Cells were subsequently lysed, and cell proteins were subjected to Western blot analysis using antibodies against (A) phospho-ERK, total ERK1/2 (B) phospho-JNK, total JNK. Corresponding densitometric analysis of Western blot experiments from pancreatic acini. The results are representative of three independent experiments. Results shown are the means + SE. * P≤ 0.05 when compared to control, + P ≤0.05 when compared to substance P (SP).
Mentions: Our data show that substance P-induced ERK1/2 and JNK activation were mediated through NK1R. We pre-treated the pancreatic acini with 1 μM of CP96345, a selective NK1R antagonist, followed by stimulation with 1 μM of substance P for 45 min. Cells were then lysed, and cell proteins were subjected to Western blot analysis. Our results, in Figure 8, demonstrate that CP96345 significantly reduced substance P-induced ERK1/2 and JNK activation in pancreatic acinar cells when compared to substance P-only treated cells.

Bottom Line: Moreover, substance P-induced ERK 1/2, JNK, NFkappaB and AP-1 activation as well as chemokine synthesis were blocked by pre-treatment with either extracellular signal-regulated protein kinase kinase 1 (MEK1) inhibitor or JNK inhibitor.In addition, substance P-induced activation of ERK 112, JNK, NFkappaB and AP-1-driven chemokine production were attenuated by CP96345, a selective NK1 R antagonist, in pancreatic acinar cells.Taken together, these results suggest that substance P-NK1 R induced chemokine production depends on the activation of MAPKs-mediated NFkappaB and AP-1 signalling pathways in mouse pancreatic acini.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, National University of Singapore, Singapore.

ABSTRACT
Substance P, acting via its neurokinin 1 receptor (NK1 R), plays an important role in mediating a variety of inflammatory processes. Its interaction with chemokines is known to play a crucial role in the pathogenesis of acute pancreatitis. In pancreatic acinar cells, substance P stimulates the release of NFkappaB-driven chemokines. However, the signal transduction pathways by which substance P-NK1 R interaction induces chemokine production are still unclear. To that end, we went on to examine the participation of mitogen-activated protein kinases (MAPKs) in substance P-induced synthesis of pro-inflammatory chemokines, monocyte chemoanractant protein-1 (MCP-I), macrophage inflammatory protein-lalpha (MIP-lalpha) and macrophage inflammatory protein-2 (MIP-2), in pancreatic acini. In this study, we observed a time-dependent activation of ERK1/2, c-Jun N-terminal kinase (JNK), NFkappaB and activator protein-1 (AP-1) when pancreatic acini were stimulated with substance P. Moreover, substance P-induced ERK 1/2, JNK, NFkappaB and AP-1 activation as well as chemokine synthesis were blocked by pre-treatment with either extracellular signal-regulated protein kinase kinase 1 (MEK1) inhibitor or JNK inhibitor. In addition, substance P-induced activation of ERK 112, JNK, NFkappaB and AP-1-driven chemokine production were attenuated by CP96345, a selective NK1 R antagonist, in pancreatic acinar cells. Taken together, these results suggest that substance P-NK1 R induced chemokine production depends on the activation of MAPKs-mediated NFkappaB and AP-1 signalling pathways in mouse pancreatic acini.

Show MeSH
Related in: MedlinePlus