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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-induced ERK1/2 and JNK cross activate NFκ B and AP-1 (c-Jun). Freshly isolated pancreatic acini, obtained from three mice, were pre-incubated with either MEK1 inhibitor PD98059 or JNK inhibitor SP600125 for 1 hr followed by stimulation with 1 μM substance P for 45 min. Acini were separated from incubation medium by centrifugation. The pellet (acini) was used for extraction and detection of (A) NFκB activation and (B) AP-1 (c-Jun) activation. 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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fig07: Substance P-induced ERK1/2 and JNK cross activate NFκ B and AP-1 (c-Jun). Freshly isolated pancreatic acini, obtained from three mice, were pre-incubated with either MEK1 inhibitor PD98059 or JNK inhibitor SP600125 for 1 hr followed by stimulation with 1 μM substance P for 45 min. Acini were separated from incubation medium by centrifugation. The pellet (acini) was used for extraction and detection of (A) NFκB activation and (B) AP-1 (c-Jun) activation. 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: Pancreatic acini were pre-treated with either PD98059 or SP600125 followed by stimulation with 1 μM substance P for 45 min. As shown in Figure 7A, PD98059 given at doses 10 μM, 30 μM, 50 μM and 100 μM significantly blocked AP-1 (c-Jun) activation. SP600125 attenuated NFκB activation at a concentration of 25 μM and above, as shown in Figure 7B. The negative control in which pancreatic acini were pre-treated with either 10 μM of PD98059 or 10 μM or even 25 μM of SP600125 (the doses sufficient to block substance P-mediated activation) for 1 hr followed by stimulation with placebo for 45 min had no significant effect on the activation of AP-1 (c-Jun) and NFκB, respectively, when compared to unstimulated controls (data not shown). Our data suggest that substance P-induced chemokine production also takes place through ERK1/2 mediated AP-1 (c-Jun) activation and JNK- mediated NFκB activation. These results imply that there is a cross-talk between the two classical pathways, ERK1/2-NFκB and JNKAP-1 (c-Jun), to induce synthesis of chemokines MCP-1, MIP-1α and MIP-2 in pancreatic acini.


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-induced ERK1/2 and JNK cross activate NFκ B and AP-1 (c-Jun). Freshly isolated pancreatic acini, obtained from three mice, were pre-incubated with either MEK1 inhibitor PD98059 or JNK inhibitor SP600125 for 1 hr followed by stimulation with 1 μM substance P for 45 min. Acini were separated from incubation medium by centrifugation. The pellet (acini) was used for extraction and detection of (A) NFκB activation and (B) AP-1 (c-Jun) activation. 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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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4401295&req=5

fig07: Substance P-induced ERK1/2 and JNK cross activate NFκ B and AP-1 (c-Jun). Freshly isolated pancreatic acini, obtained from three mice, were pre-incubated with either MEK1 inhibitor PD98059 or JNK inhibitor SP600125 for 1 hr followed by stimulation with 1 μM substance P for 45 min. Acini were separated from incubation medium by centrifugation. The pellet (acini) was used for extraction and detection of (A) NFκB activation and (B) AP-1 (c-Jun) activation. 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: Pancreatic acini were pre-treated with either PD98059 or SP600125 followed by stimulation with 1 μM substance P for 45 min. As shown in Figure 7A, PD98059 given at doses 10 μM, 30 μM, 50 μM and 100 μM significantly blocked AP-1 (c-Jun) activation. SP600125 attenuated NFκB activation at a concentration of 25 μM and above, as shown in Figure 7B. The negative control in which pancreatic acini were pre-treated with either 10 μM of PD98059 or 10 μM or even 25 μM of SP600125 (the doses sufficient to block substance P-mediated activation) for 1 hr followed by stimulation with placebo for 45 min had no significant effect on the activation of AP-1 (c-Jun) and NFκB, respectively, when compared to unstimulated controls (data not shown). Our data suggest that substance P-induced chemokine production also takes place through ERK1/2 mediated AP-1 (c-Jun) activation and JNK- mediated NFκB activation. These results imply that there is a cross-talk between the two classical pathways, ERK1/2-NFκB and JNKAP-1 (c-Jun), to induce synthesis of chemokines MCP-1, MIP-1α and MIP-2 in pancreatic acini.

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